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Sample records for apoptosis-regulating genes bax

  1. The Impact of Adenosine Fast Induction of Myocardial Arrest during CABG on Myocardial Expression of Apoptosis-Regulating Genes Bax and Bcl-2

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    Ahmed Shalaby

    2009-01-01

    Full Text Available Background. We studied the effect of fast induction of cardiac arrest with denosine on myocardial bax and bcl-2 expression. Methods and Results. 40 elective CABG patients were allocated into two groups. The adenosine group (n=20 received 250 μg/kg adenosine into the aortic root followed by blood potassium cardioplegia. The control group received potassium cardioplegia in blood. Bcl-2 and bax were measured. Bax was reduced in the postoperative biopsies (1.38 versus 0.47, P=.002 in the control group. Bcl-2 showed a reducing tendency (0.14 versus 0.085, P=.07. After the adenosine treatment, the expression of both bax (0.52 versus 0.59, P=.4 and bcl-2 (0.104 versus 0.107, P=.4 remained unaltered after the operation. Conclusion. Open heart surgery is associated with rapid reduction in the expression of apoptosis regulating genes bax and bcl-2. Fast Adenosine induction abolished changes in their expression.

  2. Expression of apoptosis-regulating genes in the rat prostate following botulinum toxin type a injection

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    Gorgal Tiago

    2012-01-01

    Full Text Available Abstract Background Onabotulinumtoxin A (OnabotA injection has been investigated as a novel treatment for benign prostatic enlargement caused by benign prostatic hyperplasia. An OnabotA - induced volume reduction caused by sympathetic fibers impairment has been proposed as a potential mechanism of action. Our aim was to investigate the expression of apoptosis-regulating proteins in the rat prostate following OnabotA intraprostatic injection. Methods Adult Wistar rats were injected in the ventral lobes of the prostate with 10 U of OnabotA or saline. A set of OnabotA-injected animals was further treated with 0.5 mg/kg of phenylephrine (PHE subcutaneously daily. All animals were sacrificed after 1 week and had their prostates harvested. Immunohistochemical staining was performed for Bax, Bcl-xL and caspase-3 proteins and visualized by the avidin-biotin method. The optical density of the glandular cells was also determined, with measurement of differences between average optical densities for each group. Results Saline-treated animals showed intense epithelial staining for Bcl-xL and a faint labelling for both Bax and Caspase-3. OnabotA-treated rats showed a reduced epithelial staining of Bcl-xL and a consistently increased Bax and Caspase-3 staining when compared with saline-treated animals. PHE-treated animals showed a stronger Bcl-xL staining and reduced staining of both Bax and Caspase-3 when compared to the OnabotA group. Mean signal intensity measurements for each immunoreaction confirmed a significant decrease of the signal intensity for Bcl-xL and a significant increase of the signal intensity for Bax and Caspase 3 in OnabotA-injected animals when compared with the control group. In OnabotA+PHE treated animals mean signal intensity for Bcl-xL, Bax and Caspase 3 immunoreactions was identical to that of the control animals. Conclusions These results support the hypothesis that OnabotA activates apoptotic pathways in the rat prostate through a

  3. Shallow needling on Neiguan and Gongsun acupoints in rats with brain ischemia Changes in apoptosis regulating genes and vasoactive substances

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    Yushan Fan; Caijiao Zhao; Wei Huang; Yan Luo; Yong Pang; Huangsha Fan; Xingui Wu; Jinsheng Wang

    2011-01-01

    For the treatment of brain ischemia using acupuncture, the needle is predominantly inserted into muscular layers and deep tissue. However, few studies have investigated the outcomes of shallow needling. The present study established middle cerebral artery occlusion models in rats using the thrombosis method. Shallow needling and conventional needling at the bilateral Neiguan (PC 6) and Gongsun (SP 4) acupoints improved neurological function of middle cerebral artery occlusion rats, increased the expression of the anti-apoptotic Bcl-2, inhibited the expression of the pro-apoptotic Bax, and reduced the expression of the vasoactive substances nitric oxide synthase and endothelin-1. However, these changes were more pronounced in the shallow needling group, indicating that shallow needling is more effective in inhibiting brain ischemic injury.

  4. THE EXPRESSION AND CLINICAL VALUE OF APOPTOSIS CONTROL GENE Bcl-2 AND Bax IN BREAST CANCER

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    ZHENG Jun; YAO Zhen-xiang; ZHANG Jing

    1999-01-01

    Objective: To study the expression and clinical value of apoptosis control gene bcl-2 and bax in breast cancer.Methods: Protein bax and bcl-2 in 41 breast cancers obtained from operations in our hospital in 1996 were detected using ABC immunohistochemical stain assay and compared with 10 cases with normal breast tissues.Results: The positive rate of bax in normal breast tissue was 90% and in breast cancer was 59%, with a significant statistical difference between them (P<0.05), but there was no statistical difference in bcl-2 protein expression. Among the 41 breast cancer, the group with lymph node metastasis (21 cases) had obviously low bax expression (43%) and high bcl-2 expression (76%), showing significant difference to the group without lymph node metastasis (P<0.05).Conclusion: The antiapoptosis function of bcl-2 was stronger than bax in breast cancer. Protein bax and bcl-2 assay may be useful in understanding the biological behaviors of breast cancer.

  5. Analysis of opa1 isoforms expression and apoptosis regulation in autosomal dominant optic atrophy (ADOA) patients with mutations in the opa1 gene.

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    Formichi, Patrizia; Radi, Elena; Giorgi, Eleonora; Gallus, Gian Nicola; Brunetti, Jlenia; Battisti, Carla; Rufa, Alessandra; Dotti, Maria Teresa; Franceschini, Rossella; Bracci, Luisa; Federico, Antonio

    2015-04-15

    Autosomal dominant optic atrophy (ADOA) is a hereditary optic neuropathy characterized by bilateral symmetrical visual loss, decrease in retinal ganglion cells and a loss of myelin within the optic nerve. ADOA is associated to mutations in Optic atrophy 1 gene (OPA1), which encodes a mitochondrial protein involved in cristae remodeling, maintenance of mitochondrial membrane integrity, mitochondrial fusion and apoptosis regulation. We thus evaluated the rate of apoptosis and the expression levels of OPA1 isoforms in ADOA and control cells. Peripheral blood lymphocytes from eight patients with OPA1 mutation and age matched controls were cultivated both in basal conditions or with 2-deoxy-D-ribose, a reducing sugar that induces apoptosis through oxidative stress. Apoptosis was analyzed by flow cytometry, phosphatidylserine translocation, mitochondrial membrane depolarization and caspase 3 activation. We also analyzed the expression levels of OPA1 isoforms in ADOA and control cells cultured with and without 2-deoxy-D-ribose. We showed an increased percentage of apoptotic cells in ADOA patients compared to controls, both in basal culture conditions and after 2-deoxy-D-ribose treatment. This suggested a great susceptibility of ADOA cells to oxidative stress and a strong correlation between OPA1 protein dysfunctions and morphological-functional alterations to mitochondria. Moreover OPA1 protein expression was significantly decreased in lymphocytes from the ADOA patients after 2-deoxy-D-ribose treatment, implying a great sensitivity of the mutated protein to free radical damage. Concluding, we could confirm that oxidative stress-induced apoptosis may play a key role in the pathophysiological process bringing to retinal ganglion cells degeneration in ADOA. PMID:25796301

  6. Effect of normothermic liver ischemic preconditioning on the expression of apoptosis-regulating genes C-jun and Bcl-XL in rats

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    Guo-Huang Hu; Xin-Sheng Lü

    2005-01-01

    AIM: To explore the expression of apoptosis-regulatinggenes C-jun and Bcl-XL after normothermic liver ischemic preconditioning and its protective effect on hepatocytes in the rat.METHODS: Wistar rats are randomly divided into sham operation group (S group, n = 10), ischemic reperfusion group (IR group, n = 10) and ischemic preconditioning group (IP group, n = 10). After dissection of the hepatoduodenal ligament in S group, and after 30-min reperfusion in IR group and in IP group, the samples of liver tissue were taken for studying the hepatocellular apoptosis, theexpressions of C-jun mRNA, Bcl-XL mRNA and their proteins, and morphologic changes at 0, 3, 6, 20 h. Meanwhile the venous blood samples were drawn at 3, 6 and 20 h for testing ALT, AST and LDH.RESULTS: The levels of ALT, AST and LDH in IR group and IP group were significantly higher than those in S group. Hepatocellular apoptosis was significantly increased in both IR group and IP group, especially in IR group.Expressions of C-jun mRNA and protein were significantly increased in IR group compared with those in both IP group and S group, but no significant difference between IP group and S group (P>0.05). Expressions of Bcl-XL mRNA and protein in IR group and S group were not significant (P>0.05), but were significantly increased in IP group compared with those in both S group and IR group. Patch necrosis of hepatocytes because of severe injury could be seen in IR group microscopically, and the ultrastructural changes were irreversible. Meanwhile in IP group, no hepatocellular necrosis occurred, and the ultrastructural changes were reversible because of mild injury. CONCLUSION: (1) IP can protect the rat liver from normothermic IR injury by modulation of the expressionof apoptosis-regulating genes C-jun and Bcl-XL; (2) IR injury may activate the apoptosis of hepatocytes by increasing the expression of apoptosis-inducing gene C-jun; (3) IP may prohibit the apoptosis of hepatocytes by increasing the

  7. Effect of Exercise Training on Bcl-2 and Bax Gene Expression in the Rat Heart

    OpenAIRE

    Jafari; Pourrazi; Nikookheslat; Baradaran

    2015-01-01

    Background Apoptosis or programmed cell death plays an important role in the development of cardiovascular diseases, particularly heart failure. Current evidence suggests that exercise training may alter apoptosis-related signaling in sensitive somatic tissues such as the myocardium. Objectives The aim of this study was to assess the effect of exercise training on Bcl-2 and Bax genes expression as key molecules involved in intrins...

  8. Darbepoetin alpha, a long-acting erythropoeitin derivate, does not alter LPS evoked myocardial depression and gene expression of Bax, Bcl-Xs, Bcl-XL, Bcl-2, and TNF-alpha.

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    Brendt, Peter; Frey, Ulrich; Adamzik, Michael; Schäfer, Simon T; Peters, Jürgen

    2009-01-01

    Darbepoetin alpha (DA), a long-acting erythropoietin derivative stimulating erythropoiesis, can, by antiapoptotic effects, mitigate myocardial I/R injury. We tested the hypothesis that DA treatment improves left ventricular function (LV) in LPS evoked cardiomyopathy and alters gene expression of apoptosis-regulating proteins (Bcl-XL, Bcl-2, Bax, and Bcl-Xs) and TNF-alpha. In a prospective, controlled, randomized study in Lewis rats (n = 56; 8 groups), myocardial depression was evoked by LPS administration (serotype O127:B8; 10 mg/kg, i.p.). Darbepoetin alpha or vehicle was injected either 24 h before (pretreatment) or 2 h after LPS injection (treatment). Hearts were isolated 8 h after LPS injection, perfused (Krebs-Henseleit solution) in a Langendorff apparatus, and LV developed pressure and its derivatives were measured. For gene expression analysis, real-time polymerase chain reaction of LV specimen was performed. LPS decreased LV developed pressure (-64.6 +/- 7.9 mmHg) and its derivates by more than 60% in comparison to vehicle (P Xs, Bax, and TNF-alpha, but this was not altered by DA pretreatment. Furthermore, there was no effect on Bcl-Xl and Bcl-2 expression by DA alone. Whereas proapoptotic genes of the myocardium are up-regulated in LPS-induced cardiomyopathy, neither DA pretreatment nor treatment has significant effects on LV function or gene expression. This may suggest cardiac resistance to darbepoetin in LPS-mediated sepsis.

  9. CO-EXPRESSIONS OF SURVIVIN GENE,BCL-2 AND BAX PROTEINS IN OVARIAN CARCINOMA

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    林蓓; 张淑兰; 赵长清

    2004-01-01

    Objective To characterize the cellular properties of ovarian cancer, we examined the correlation between the expression of apoptosis-related gene survivin and those of Bcl-2 and Bar proteins. Methods Expressions of survivin mRNA, and Bcl-2 and Bax proteins in 35 cases of ovarian carcinoma, 10 cases of borderline carcinoma, 10 cases of benign tumors and 10 cases of normal tissue were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry SABC method, respectively. Results Expression of survivin gene was detected in a significantly greater proportion in ovarian carcinoma and borderline carcinoma than those in benign tumors and normal tissues. Although there was no relationship between expression of survivin gene and FIGO stage, histologic grade, pathological type and lymphatic metastasis, expressions of Bcl-2 and Bar proteins were positively and negatively correlated with that of survivin gene, respectively. Conclusion Survivin may play an important role in pathogenesis of ovarian carcinoma, with a synergistic role of apoptosis-related gene Bcl-2protein and an antagonistic role of Bax protein in formation and progression of ovarian carcinoma.

  10. Low levels of Bax inhibitor-1 gene expression increase tunicamycin-induced apoptosis in human neuroblastoma SY5Y cells

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    Dan Wu; Peirong Wang; Shiyao Wang

    2012-01-01

    A human SH-SY5Y neuroblastoma cell line with a low level of Bax inhibitor-1 expression was established by lentivirus-mediated RNA interference and fluorescence-activated cell sorting. In control SH-SY5Y cells, tunicamycin treatment induced endoplasmic reticulum stress-mediated apoptosis; however, after Bax inhibitor-1 gene knockdown, cell survival rates were significantly decreased and the degree of apoptosis was significantly increased following tunicamycin treatment. In addition, chromatin condensation and apparent apoptotic phenomena, such as marginalization and cytoplasmic vesicles, were observed. Our findings indicate that Bax inhibitor-1 can delay apoptosis induced by endoplasmic reticulum stress.

  11. A single nucleotide polymorphism in the Bax gene promoter affects transcription and influences retinal ganglion cell death

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    Sheila J Semaan

    2010-03-01

    Full Text Available Pro-apoptotic Bax is essential for RGC (retinal ganglion cell death. Gene dosage experiments in mice, yielding a single wild-type Bax allele, indicated that genetic background was able to influence the cell death phenotype. DBA/2JBax+/− mice exhibited complete resistance to nerve damage after 2 weeks (similar to Bax−/− mice, but 129B6Bax+/− mice exhibited significant cell loss (similar to wild-type mice. The different cell death phenotype was associated with the level of Bax expression, where 129B6 neurons had twice the level of endogenous Bax mRNA and protein as DBA/2J neurons. Sequence analysis of the Bax promoters between these strains revealed a single nucleotide polymorphism (T129B6 to CDBA/2J at position −515. A 1.5- to 2.5-fold increase in transcriptional activity was observed from the 129B6 promoter in transient transfection assays in a variety of cell types, including RGC5 cells derived from rat RGCs. Since this polymorphism occurred in a p53 half-site, we investigated the requirement of p53 for the differential transcriptional activity. Differential transcriptional activity from either 129B6 or DBA/2J Bax promoters were unaffected in p53−/− cells, and addition of exogenous p53 had no further effect on this difference, thus a role for p53 was excluded. Competitive electrophoretic mobility-shift assays identified two DNA–protein complexes that interacted with the polymorphic region. Those forming Complex 1 bound with higher affinity to the 129B6 polymorphic site, suggesting that these proteins probably comprised a transcriptional activator complex. These studies implicated quantitative expression of the Bax gene as playing a possible role in neuronal susceptibility to damaging stimuli.

  12. Effect of Exercise Training on Bcl-2 and Bax Gene Expression in the Rat Heart

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    Jafari

    2015-10-01

    Full Text Available Background Apoptosis or programmed cell death plays an important role in the development of cardiovascular diseases, particularly heart failure. Current evidence suggests that exercise training may alter apoptosis-related signaling in sensitive somatic tissues such as the myocardium. Objectives The aim of this study was to assess the effect of exercise training on Bcl-2 and Bax genes expression as key molecules involved in intrinsic pathway of apoptosis in the rat heart. Materials and Methods This study was conducted with a two-group experimental design (animal model and sixteen three-month-old male rats were selected and randomly divided to two groups of exercise training (n = 8 and control (n = 8. Rats in the trained group participated in an exercise training program for 12 weeks (10 – 60 m min-1, 24 – 33 min d-1, 15%. The rat hearts were removed forty-eight hours after the last training session. RNA extraction and synthesis of cDNA was done, and Bax and Bcl2 genes expression was analyzed through the Real Time-Polymerase Chain Reaction (RT-PCR. Kolmogorov-Smirnov and independent t-test were applied for statistical analysis of the data (P 0.05. However, Bcl2 expression was higher in the trained group compared to the control group (11%. Conclusions In general, it seems that three-month exercise training was effective in reducing cardiac mitochondrial pro-apoptotic protein. However, considering the results of the Bcl2 gene expression, more researches are needed to identify effects of exercise trainings on indices of myocardial apoptosis.

  13. Erythropoietin can promote survival of cerebral cells by downregulating Bax gene after traumatic brain injury in rats

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    Liao Z

    2009-01-01

    Full Text Available Background : Traumatic brain injury (TBI is an important cause of adult mortality and morbidity. Erythropoietin (Epo has been shown to promote the viability of cerebral cells by upregulating Bcl-2 gene; however, Epo may exert its antiapoptotic effect via the differential regulation of the expression of genes involved in the apoptotic process. Aim : The present study examined the neuroprotective effect of Epo as a survival factor through the regulation of the Bax. Materials and Methods : Wistar rats were randomly divided into three groups: Recombinant human EPO treated (rhEPO TBI, vehicle-treated TBI, and sham-operated. Traumatic brain injury was induced by the Feeney free-falling model. Rats were killed 5, 12, 24, 72, 120, or 168 h after TBI. Regulation of Bcl-2 was detected by reverse transcription-polymerase chain reaction (RT-PCR, western blotting and immunofluorescence. Results : Bax mRNA and protein levels were lower in the rhEPO-treated rat brains than in the vehicle-treated rat brains. Induction of Bax expression peaked at 24 h and remained stable for 72-120 h in vehicle-treated rat brains, whereas induction of Bax expression was only slightly elevated in rhEPO-treated rat brains. The number of TdT-mediated dUTP Nick-End Labeling(TUNEL-positive cells in the rhEPO-treated rat brains was far fewer than in the vehicle-treated rat brains. Conclusions : Epo exerts neuroprotective effect against traumatic brain injury via reducing Bax gene expression involved in inhibiting TBI-induced neuronal cell death.

  14. Decoding c-Myc networks of cell cycle and apoptosis regulated genes in a transgenic mouse model of papillary lung adenocarcinomas.

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    Ciribilli, Yari; Singh, Prashant; Spanel, Reinhard; Inga, Alberto; Borlak, Jürgen

    2015-10-13

    The c-Myc gene codes for a basic-helix-loop-helix-leucine zipper transcription factor protein and is reported to be frequently over-expressed in human cancers. Given that c-Myc plays an essential role in neoplastic transformation we wished to define its activity in lung cancer and therefore studied its targeted expression to respiratory epithelium in a transgenic mouse disease model. Using histological well-defined tumors, transcriptome analysis identified novel c-Myc responsive cell cycle and apoptosis genes that were validated as direct c-Myc targets using EMSA, Western blotting, gene reporter and ChIP assays.Through computational analyses c-Myc cooperating transcription factors emerged for repressed and up-regulated genes in cancer samples, namely Klf7, Gata3, Sox18, p53 and Elf5 and Cebpα, respectively. Conversely, at promoters of genes regulated in transgenic but non-carcinomatous lung tissue enriched binding sites for c-Myc, Hbp1, Hif1 were observed. Bioinformatic analysis of tumor transcriptomic data revealed regulatory gene networks and highlighted mortalin and moesin as master regulators while gene reporter and ChIP assays in the H1299 lung cancer cell line as well as cross-examination of published ChIP-sequence data of 7 human and 2 mouse cell lines provided strong evidence for the identified genes to be c-Myc targets. The clinical significance of findings was established by evaluating expression of orthologous proteins in human lung cancer. Taken collectively, a molecular circuit for c-Myc-dependent cellular transformation was identified and the network analysis broadened the perspective for molecularly targeted therapies.

  15. Identification of Novel Pepper Genes Involved in Bax- or INF1-Mediated Cell Death Responses by High-Throughput Virus-Induced Gene Silencing

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    Jeong Hee Lee

    2013-11-01

    Full Text Available Hot pepper is one of the economically important crops in Asia. A large number of gene sequences, including expressed sequence tag (EST and genomic sequences are publicly available. However, it is still a daunting task to determine gene function due to difficulties in genetic modification of a pepper plants. Here, we show the application of the virus-induced gene silencing (VIGS repression for the study of 459 pepper ESTs selected as non-host pathogen-induced cell death responsive genes from pepper microarray experiments in Nicotiana benthamiana. Developmental abnormalities in N. benthamiana plants are observed in the 32 (7% pepper ESTs-silenced plants. Aberrant morphological phenotypes largely comprised of three groups: stunted, abnormal leaf, and dead. In addition, by employing the combination of VIGS and Agrobacterium-mediated transient assays, we identified novel pepper ESTs that involved in Bax or INF1-mediated cell death responses. Silencing of seven pepper ESTs homologs suppressed Bax or INF1-induced cell death, five of which suppressed both cell death responses in N. benthamiana. The genes represented by these five ESTs encode putative proteins with functions in endoplasmic reticulum (ER stress and lipid signaling. The genes represented by the other two pepper ESTs showing only Bax-mediated cell death inhibition encode a CCCH-type zinc finger protein containing an ankyrin-repeat domain and a probable calcium-binding protein, CML30-like. Taken together, we effectively isolated novel pepper clones that are involved in hypersensitive response (HR-like cell death using VIGS, and identified silenced clones that have different responses to Bax and INF1 exposure, indicating separate signaling pathways for Bax- and INF1-mediated cell death.

  16. Effects of Chronic Mild Stress in Female Bax Inhibitor-1-Gene Knockout Mice

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    Sui, Zhi-Yan; Chae, Han-Jung; Huang, Guang-Biao; Zhao, Tong; Shrestha Muna, Sushma; Chung, Young-Chul

    2012-01-01

    Objective The anti-apoptotic protein Bax inhibitor-1 (BI-1) is a regulator of apoptosis linked to endoplasmic reticulum (ER) stress, and BI-1-/- mice exhibit increased sensitivity to tissue damage. The purpose of this study was to investigate the role of BI-1 in the pathogenesis of chronic mild stress (CMS)-induced depression-like behaviors in BI-1-/- mice. Methods We delivered CMS for 2 or 6 weeks in BI-1-knockout and wild-type mice. Control groups of BI-1-knockout and wild-type mice were le...

  17. The Expression of Apoptosis-Related Genes Bcl-2 and Bax Protein and Apoptosis Positivity in Cervical Carcinoma during Irradiation

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    ZHAODongli; SHIJingsen; LIMingzhong; SONGLiping; WANGShuwen

    2005-01-01

    Objective: To evaluate the apoptosis positivity, the expression of Bcl-2. bax proteins in 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By using immunohistochemical and TDT-dUTP nick end labelling techniques. 30 cases of squamous cell cervical carcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%, and 100% respectively, with the difference being significant (P<0.05); The positive rates of Bcl-2 protein before and after irradiation were 73.3% and 46.7% respectively, with the difference being significant (P<0.05): The positive rates of bax protein before and after irradiation were 86% and 100 respectively, with the difference being significant (P<0.05). Conclusion: bax and Bcl-2 protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosis induced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2 protein.

  18. Expression of bax and bcl2 Genes in MDMA-induced Hepatotoxicity on Rat Liver Using Quantitative Real-Time PCR Method through Triggering Programmed Cell Death

    OpenAIRE

    2015-01-01

    Background: 3-4methylenedioxymethamphetamine (MDMA) is a synthetic and psychoactive drug, which is known popularly as Ecstasy and has toxic effects on human organs. Objectives: Considering the potential toxic interaction, this study was performed to quantify the expression of bax and bcl2 genes in MDMA-induced hepatotoxicity on rat liver. Subsequently, we evaluated pentoxifylline as a possible protective drug on hepatotoxicity. Materials and Methods: Adult male Wistar rats weighting 250 - 300...

  19. The bcl-2, bax gene expression and apoptosis of continuous low-dose-rate irradiation on PC-3 transplanting tumor

    International Nuclear Information System (INIS)

    Objective: The aim of this study was to investigate bcl-2, bax expression and apoptosis of continuous low-dose-rate irradiation on prostate cancer (PC)-3 transplanting tumor. Methods: The expression of bcl-2 and bax associated with apoptosis between experiment and control groups were analyzed using immunohistochemistry at 48, 96 and 192 h after two 125I seed sources implanting model. The correlation between apoptosis and the ratio of bax/bcl-2 was analyzed using Bi-variable linear correlation. SPSS 11.0 was used to analyse the data. Results: The bcl-2 expression in experiment group began to down-regulated significantly after 125I seed irradiation for 48 h as compared with control(t=2.500, P=0.067), though it was not reached to statistical significance. At 96 and 192 h after irradiation, significantly low expression of bcl- 2 were noted (t=4.950, 3.464; P=0.008 and 0.026). In contrast, significantly over expression of bax was noted at 48, 96 and 192 h after 12si irradiation (t=3.334,4.025,5.292;P=0.029, 0.016 and 0.006). The apoptotic index (AI) for PC-3 at 48, 96 and 192 h after 125I irradiation were 22.3%, 21.7% and 30.7%, which was significantly higher than controls when at 96 and 192 h after 125I irradiation (P= 0.016 and 0.036). Moreover, positive correlation was noted between AI and bax/bcl-2 ratio (r=0.784, P= 0.012). Conclusion: Low-dose-rate irradiation could down-regulate the expression of bcl-2, up-regulate the expression of bax and induced PC-3 cells apoptosis. (authors)

  20. TNFα PRODUCTION AND APOPTOSIS REGULATION IN VIRAL HEPATITIS TYPE C

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    V. V. Novitsky

    2005-01-01

    Full Text Available Abstract. Chronical course of infection caused by hepatitis C virus is accompanied by increase Fas-positive lymphocytes of peripheral blood. Cultivation of agglutinin-stimulated mononuclear blood cells of patients with chronic hepatitis C revealed inhibition of apoptotic reactions of blood lymphocytes. This fact correlated with decrease in production of TNFα and accelerated synthesis of soluble receptor for this cytokine. We suggest a virus-specific influence on apoptosis regulation of target cells.

  1. Expression of protein encoded by apoptosis-associated gene p53, bcl-2, and bax in adaptive response of thymocyte apoptosis in mice induced by low dose radiation with X-rays

    International Nuclear Information System (INIS)

    Objective: To explore the regulative mechanism of apoptosis-associated gene proteins on the adaptive response of thymocyte apoptosis in mice induced by low dose radiation with X-rays. Methods: Kunming male mice were irradiated with the inductive doses (D1: 25, 50, 75, 100 and 200 mGy; dose rate: 12.5 mGy ·min-1) and the challenging dose (D2: 1.5 Gy; dose rate: 287 mGy·min-1). The time interval between D1 and D2 was 6 h. The expressive levels of thymocyte apoptosis-associated gene proteins were measured with flow cytometry. Results: As compared with the sham-irradiation, the positive percentage of thymocyte Bcl-2 protein expression decreased significantly in D2 group (P<0.05), Bax increased significantly (P<0.05), and Bcl-2/Bax decreased significantly (P<0.001); p 53 increased significantly (P<0.001). As compared with D2 group, the positive percentage of thymocyte Bcl-2 protein expression increased in varying degree in D1+ D2 group of 25-75 mGy D1, Bax decreased in varying degree, and Bcl-2/Bax increased significantly (P<0.01); p53 decreased significantly (P<0.001 or P<0.05). Conclusion: The apoptotic thymocytes in the adaptive response of thymocyte apoptosis in mice induced by irradiation with 25-75 mGy decrease significantly due to the increase of apoptosis-associated gene Bcl-2 protein expression and Bcl-2/Bax, the decrease of Bax and p53 protein expressions. (authors)

  2. Human Papillomavirus 16 E6E7 fusion gene′s impact on the expression of apoptosis regulation genes in esophageal squamous cancer cells KYSE450%HPV16 E6/E7对凋亡调控基因表达影响的研究

    Institute of Scientific and Technical Information of China (English)

    沙亚哈提·别尔克哈之; 李卉; 吉别克·瓦提别克; 刘伊宁; 李晓苗; 来雯婷; 美丽吾尔提·达吾列提汗; 谌宏鸣; 李惠武

    2014-01-01

    目的:探讨 HPV16E6/E7对 KYSE450细胞中凋亡调控因子 Bcl-2和 Bad 表达的影响。方法采用瞬时转染技术将 HPV1E6/E7融合基因的真核表达载体转染食管癌细胞株 KYSE450中,分组用 TSA(HDACi)处理后,收集 RNA,应用反转录-聚合酶链反应(reverse transcription polymerase chain reaction RT-PCR)技术检测Bcl-2、Bad 的 mRNA 表达情况。结果(1)用 RT-PCR 检测转染后的结果为阳性。(2)Bad mRNA 在食管癌细胞KYSE450中表达较低,E6E7过表达对细胞内 Bad 基因 mRNA 水平无影响,各转染组之间没有差异。(3)Bcl-2 mRNA 在食管癌细胞 KYSE450中均有表达,E6/E7对 Bcl-2有轻度调控作用。(4)HDAC 抑制剂 TSA 处理后, TSA 明显增加了 E6对 Bcl-2基因表达的诱导调控作用。结论KYSE450细胞中 E6和 E7基因促进 Bcl-2在转录水平上的表达,而对 Bad 的表达无明显的影响。在病毒感染鳞状上皮细胞后早期表达致癌基因 E6、E7可能诱导凋亡调控因子 Bcl-2在转录水平上的表达。%Objective To investigate the human papillomavirus 16 (HPV16 )E6E7′s impact on the expres-sion of apoptosis regulation genes Bcl-2 and Bad in esophageal squamous cancer cell line KYSE-450.Meth-ods E6E7 fusion eukaryotic expression vector was transfected into the esophageal cell lines KYSE450 by transient transfection technique,and grouped with TSA(HDACi)treatment;the expressions of Bcl-2 and Bad were detected by reverse transcription polymerase chain reaction RT-PCR.Results (1)The result af-ter transfection was positive by RT-PCR.(2)There was low expression of Bad gene in KYSE-450 cell lines;E6E7overexpression had no effect on Bad mRNA and there was no any difference between the groups.(3)Bcl-2 gene was expressed in KYSE-450 cell lines,suggesting that Bcl-2 was slightly regulated by E6E7.(4)Treatment after TSA(HDACi),which significantly increase the regulation of E6 on the Bcl-2 expres-sion.Conclusion E6,E7

  3. Lycopene modulates cholinergic dysfunction, Bcl-2/Bax balance, and antioxidant enzymes gene transcripts in monosodium glutamate (E621) induced neurotoxicity in a rat model.

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    Sadek, Kadry; Abouzed, Tarek; Nasr, Sherif

    2016-04-01

    The effect of monosodium glutamate (MSG) on brain tissue and the relative ability of lycopene to avert these neurotoxic effects were investigated. Thirty-two male Wistar rats were distributed into 4 groups: group I, untreated (placebo); group II, injected with MSG (5 mg·kg(-1)) s.c.; group III, gastrogavaged with lycopene (10 mg·kg(-1)) p.o.; and group IV received MSG with lycopene with the same mentioned doses for 30 days. The results showed that MSG induced elevation in lipid peroxidation marker and perturbation in the antioxidant homeostasis and increased the levels of brain and serum cholinesterase (ChE), total creatine phosphokinase (CPK), creatine phosphokinase isoenzymes BB (CPK-BB), and lactate dehydrogenase (LDH). Glutathione S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) activities and gene expression were increased and glutathione content was reduced in the MSG-challenged rats, and these effects were ameliorated by lycopene. Furthermore, MSG induced apoptosis in brain tissues reflected in upregulation of pro-apoptotic Bax while lycopene upregulated the anti-apoptotic Bcl-2. Our results indicate that lycopene appears to be highly effective in relieving the toxic effects of MSG by inhibiting lipid peroxidation and inducing modifications in the activity of cholinesterase and antioxidant pathways. Interestingly, lycopene protects brain tissue by inhibiting apoptosis signaling induced by MSG. PMID:26900785

  4. Expression and significance of Bcl-2 gene and Bax gene in lung cancer%凋亡相关基因bcl-2和bax在肺癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    郭雷; 王福昌; 杨五计

    2001-01-01

    To study the relationship between expression of bcl-2 and baxgene and lung cancer.The bcl-2 and bax gene expression in 51 lung cancer tissues and tissues near cancer were deteeted by immunohistochemical SP method.The positive degree of bcl-2 gene expression in lung cancer was higher than that in tissues near cancer.The positive degree of bax gene expression in lung cancer was lower than that in tissues near cancer.Both differences were very significant(P<0.01).There was no correlations between the immunoreactivity of bcl-2 and bax gene and histology,gender,smoking history,clinical subtypes,stage and lymphatic metastasis(P>0.05).It suggests that bcl-2 and bax gene may play an important role in tumorgenesis and tumor development.The detection and determination of bcl-2 and bax gene in preinvasive lesions may contribute to the early diagnosis of lung cancer.Some drugs can down-regulate bcl-2 expression and speed apoptosis so as to improve the sensitivity to chemical therapy and radiation.%为探讨B细胞淋巴瘤/白血病-2(bcl-2)基因及其同源类似物bax基因的表达与肺癌发生的关系,应用免疫组化SP法检测了51例肺癌患者的肺癌标本及其癌旁组织中bcl-2、bax的表达。结果显示,bcl-2在肺癌中染色比在癌旁组织中深,bax在肺癌中染色比在癌旁组织中浅,两者之间均有极显著差异(P<0.01);bcl-2、bax表达与肺癌的组织分型、患者的性别、吸烟史、临床分型、分期及淋巴结转移无关(P>0.05)。提示:①bcl-2、bax在肺癌发生中具有重要作用。②在癌前病变组织中检测并比较bcl-2和bax的表达,有利于肺癌的早期诊断。③可通过药物使bcl-2表达降低,促进凋亡,提高患者对放、化疗的敏感性。

  5. Silver Nanoparticles Biosynthesized Using Achillea biebersteinii Flower Extract: Apoptosis Induction in MCF-7 Cells via Caspase Activation and Regulation of Bax and Bcl-2 Gene Expression

    Directory of Open Access Journals (Sweden)

    Javad Baharara

    2015-02-01

    Full Text Available Silver nanoparticles (Ag-NPs, the most popular nanoparticles, possess unique properties. Achillea biebersteinii is a plant of the Asteraceae family rich in active antitumor components. The aim of this research was the characterization and investigation of the cytotoxic properties of Ag-NPs synthesized using A. biebersteinii flower extract, on a human breast cancer cell line. The Ag-NPs were synthesized after approximately 180 min of reaction at 40 °C, then they were characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR, transmission electron microscopy (TEM and dynamic light scattering (DLS. The anti-apoptosis effect of Ag-NPs on the MCF-7 cell line was investigated by MTT assay, DAPI and acridine orange staining and caspase activity. The transcriptional expression of bax, bcl-2, caspase-3, -8 and -9 were also evaluated by RT-PCR. The TEM images revealed that the Ag-NPs morphology had a different shape. The DLS indicated that the average hydrodynamic diameter of the biosynthesized Ag-NPs was around 12 nm. By UV-visible spectroscopy the strongest absorbance peak was observed at 460 nm. The FTIR results also showed interaction between the plant extract and Ag-NPs due to the similarity in the peak patterns. The EDS results showed that Ag-NPs display an absorption peak at 3 keV, indicating the presence of the element silver. The Ag-NPs caused a dose-dependent decrease in cell viability, fragmentation in nucleic acid, inhibited the proliferation and induction of apoptosis on MCF-7 by suppressing specific cell cycle genes, and simulation programmed cell dead genes. Further investigation is required to establish the potential of this novel and promising approach in cancer therapy.

  6. The apoptotic response in HCT116BAX-/- cancer cells becomes rapidly saturated with increasing expression of a GFP-BAX fusion protein

    International Nuclear Information System (INIS)

    Many chemotherapeutic agents promote tumor cell death by activating the intrinsic pathway of apoptosis. Intrinsic apoptosis involves permeabilization of the mitochondrial outer membrane and the release of cytochrome c, a process that is controlled by proteins of the BCL2 gene family. Chemoresistance is often associated with abnormalities in concentrations of BCL2 family proteins. Although stoichiometirc interactions between anti-apoptotic and BH3-only BCL2 family proteins have been well documented as affecting cell death, the association between changes in BAX concentration and intrinsic apoptosis are poorly understood. Exogenous GFP-murine Bax fusion constructs were transfected into BAX-deficient HCT116 cells. To titrate the expression of the fusion protein, GFP-BAX was cloned into a tetracycline sensitive expression cassette and cotransfected with a plasmid expressing the rtTA transcription factor into HCT116BAX-/- cells. Linear expression of the fusion gene was induced with doxycycline and monitored by quantitative PCR and immunoblotting. Cell death was assayed by DAPI staining cells after exposure to indomethacin, and scoring nuclei for condensed chromatin and fragmented nuclei. HCT116BAX-/- cells were resistant to indomethacin, but susceptibility could be recovered in cells expressing a GFP-BAX fusion protein. Titration of GFP-BAX expression revealed that the concentration of BAX required to induce a saturating apoptosis response from baseline, was rapidly achieved. Increased levels of GFP-BAX were unable to stimulate higher levels of cell death. Examination of GFP-BAX distribution before and after indomethacin treatment indicated that BAX protein did not form aggregates when present at sub-lethal concentrations. Within the limitations of this experimental system, BAX-dependent apoptosis in HCT116 cells exhibits an all-or-none response depending on the level of BAX protein present. The lack of BAX aggregation at sub-saturation levels suggests that the

  7. 细胞凋亡相关基因Bcl-2及Bax在骨肉瘤中的表达与自下而上质量的关系%Expression of apoptosis related gene Bcl 2 and Bax in osteosarcoma and their relationship with the prognosis

    Institute of Scientific and Technical Information of China (English)

    黄鲁豫; 刘建; 王臻; 吕荣

    2002-01-01

    Objective Apoptosis related gene Bcl 2 and Bax in osteosarcoma patients with different clinical appearance were being studied to analyze the prognosis of the patients. Method The cases were divided into two different groups according to the results of the follow up.33 cases in high risk group and 18 cases in low risk group. Expression of Bcl 2 and Bax were immunohistochemically stained by ABC method. Result Positive expression rate of Bcl 2 was 61% in high risk group (20/23) and 33% in low risk group (1/8). Positive expression of Bax was 22% in high risk group (6/27) and 67% in low risk group(12/18).Conclusion Expression of Bcl 2 and Bax was related to the prognosis of osteosarcoma. Positively expressed Bcl 2 in osteosarcoma cells may indicate bad prognosis. If Bax is highly expressed in osteosarcoma cells, this may indicated a good prognosis.

  8. Microsatellite instability and frameshift mutations in BAX and transforming growth factor-beta RII genes are very uncommon in acute lymphoblastic leukemia in vivo but not in cell lines.

    Science.gov (United States)

    Molenaar, J J; Gérard, B; Chambon-Pautas, C; Cavé, H; Duval, M; Vilmer, E; Grandchamp, B

    1998-07-01

    Mutations in the DNA mismatch repair (MMR) system lead to an instability of simple repetitive DNA sequences involved in several cancer types. This instability is reflected in a high mutation rate of microsatellites, and recent studies in colon cancer indicate that defects in MMR result in frequent frameshift mutations in mononucleotide repeats located in the coding regions of BAX and transforming growth factor-beta (TGF-beta) receptor genes. Circumstantial evidence suggests that the MMR defect may be involved in some lymphoid malignancies, although several allelotype analyses have concluded on the low level of microsatellite instability in acute lymphoblastic leukemias. To further evaluate the implication of MMR defects in leukemogenesis, we have studied a series of 98 children with acute lymphoblastic leukemia and 14 leukemic cell lines using several indicators of MMR defects. Microsatellite markers were compared between blast and normal DNA from the same patients and mutations were sought in mononucleotide repeat sequences of BAX and TGF-beta receptor II (TGF-beta RII). The absence of microsatellite instability (MI) and the absence of mutations in the genes examined from patient's leukemic cells contrasted with the observation that half of the cell lines displayed a high degree of MI and that three of seven of these mutator cell lines harbored mutations in BAX and/or TGF-beta RII. From these results we conclude that MMR defects are very uncommon in freshly isolated blasts but are likely to be selected for during the establishment of cell lines.

  9. Deregulated expression of A1, Bcl-2, Bcl-xL, and Mcl-1 antiapoptotic proteins and Bid, Bad, and Bax proapoptotic genes in polycythemia vera patients

    Directory of Open Access Journals (Sweden)

    Elainy Patricia Lino Gasparotto

    2011-12-01

    Full Text Available Apoptosis deregulation might have a role in the pathophysiology of polycythemia vera (PV. This study evaluated Bcl-2 molecule expression in CD34+ cells and leukocytes in 12 PV patients. Gene expression was investigated by real time PCR using SybrGreen Quantitect kit and protein expression was evaluated by western-blotting. JAK2 V617F mutation was detected according to Baxter et al (2005. CD34+ cells from PV patients presented higher levels of A1 and Mcl-1 expression (median: 22.6 and 5.2, respectively in comparison with controls (0.9 and 0.5, p=0.004 and p=0.020; while Bcl-2 and Bcl-xL expression decreased in PV patients (0.18 and 1.19 compared with controls (1.39 and 2.01, p=0.006 and p=0.020. CD34+ cells in PV patients showed an elevated Bid expression (14.4 in comparison with healthy subjects (1.0; p=0.002. Patients' leukocytes showed an A1 augmentation (7.41, p=0.001 and a reduced expression of Bax (0.19; p=0.040 and Bad (0.2; p=0.030. There was no correlation between JAK2 V617F allele burden and molecular expression. PV patients showed alterations in Bcl-2 members' expression, which may interfere with control of apoptotic machinery and contribute to disease pathogenesis.A desregulação da apoptose parece participar da fisiopatologia da policitemia vera (PV. Este estudo avaliou a expressão das moléculas da família Bcl-2 em células hematopoéticas CD34 + e leucócitos de 12 pacientes com PV. Foram realizados: a quantificação da expressão gênica por PCR em tempo real utilizando kit Sybrgreen Quantitect, avaliação da expressão de proteínas por western-blot e detecção da mutação JAK2 V617F segundo Baxter et al. (2005. Células CD34 + dos pacientes com PV apresentaram maior expressão de A1 e Mcl-1 (mediana: 22,6 e 5,2, respectivamente em comparação com controles (0,9 e 0,5, p = 0,004 e p = 0,020 e expressão de Bcl-2 e Bcl-xL diminuída nestes pacientes (0,18 e 1,19 em relação aos controles (1,39 e 2,01, p = 0,006 e p = 0

  10. bax和p53基因共转染对人舌癌细胞增 殖和凋亡的影晌%Effect on Proliferation and Apoptosis of Human Lingual Carc inoma Cells Cotransfected by bax and p53 Genes

    Institute of Scientific and Technical Information of China (English)

    马英红; 杨绍华; 陈俊杰; 高家让; 彭文珍; 王若菡

    2001-01-01

    Objective To investigate th e effects on prolife ration andapoptosis of human cancerous cells cotransfected by bax apoptosis-in ducing gene and p53 tumor suppressor gene. Methods The chi meric gene pSV-CIP-bax -CAT was constructed in which bax gene was flanked upstream by a 217bp fragment (+822~+1093) of the first intron and a 317bp promoter fragment of human a 1(Ⅰ) colla gen gene . Human lingual carcinoma cell line Tca 8113 (LCC) in culture was respe ctively transfected with pSV-CIP-bax-CAT, and cotransfected with pSV-p53-CA T and pSV-CIP-bax-CAT by using the transfection reagent DOSPER. Res ults Immuno- slot blot and ELISA demonstrated that the expression of bax and p53 genes incr eased remarkably in the transfected and cotransfected LCC, compared with the con trols(LCC transfected with pSV-CIP-CAT and LCC).MTT colorimetric assay,TUNE L fluorecence microscopy and flow cytometry showed that foreign bax or p53 gene inhibited the LCC growth and induced apoptosis(23.9% and 26.1% of inhibitory ra te by bax gene and p53 plus bax genes respectively). Conclusion The ectopic expression of the bax gene in the LCC promoted by the cis-act ing elements of human a1(Ⅰ)collagen gene has obviously synergistic effect on th e proliferation and apoptosis of the LCC cotransfected with p53 gene plus bax gene for 48 hours.%目的 探讨诱导凋亡的bax基因和p53抑癌基因共转染对人癌细胞增殖和凋亡的作用。方法 构建pSV-CIP-bax-CAT嵌合基因。在其bax基因上游含人a1(Ⅰ)型胶原基因的第一内含子217bp片段(+822~+1093)和317bp启动子片段。经阳离子转染试剂DOSPER介导,分别用pSV-CIP-bax-CAT转染、pSV-CIP-bax-CAT和pSV-p53-CAT共转染培养的人舌癌Tca8113细胞系(LCC)。结果 免疫狭缝印迹和ELISA检测证实,转染组和共转染组比对照的bax和p53基因表达量明显增加。MTT显色分析、TUNEL荧光显微镜和流式细胞仪监测结果显示,bax基因或p53基因均具抑

  11. Conformational Heterogeneity of Bax Helix 9 Dimer for Apoptotic Pore Formation

    Science.gov (United States)

    Liao, Chenyi; Zhang, Zhi; Kale, Justin; Andrews, David W.; Lin, Jialing; Li, Jianing

    2016-07-01

    Helix α9 of Bax protein can dimerize in the mitochondrial outer membrane (MOM) and lead to apoptotic pores. However, it remains unclear how different conformations of the dimer contribute to the pore formation on the molecular level. Thus we have investigated various conformational states of the α9 dimer in a MOM model — using computer simulations supplemented with site-specific mutagenesis and crosslinking of the α9 helices. Our data not only confirmed the critical membrane environment for the α9 stability and dimerization, but also revealed the distinct lipid-binding preference of the dimer in different conformational states. In our proposed pathway, a crucial iso-parallel dimer that mediates the conformational transition was discovered computationally and validated experimentally. The corroborating evidence from simulations and experiments suggests that, helix α9 assists Bax activation via the dimer heterogeneity and interactions with specific MOM lipids, which eventually facilitate proteolipidic pore formation in apoptosis regulation.

  12. Conformational Heterogeneity of Bax Helix 9 Dimer for Apoptotic Pore Formation.

    Science.gov (United States)

    Liao, Chenyi; Zhang, Zhi; Kale, Justin; Andrews, David W; Lin, Jialing; Li, Jianing

    2016-01-01

    Helix α9 of Bax protein can dimerize in the mitochondrial outer membrane (MOM) and lead to apoptotic pores. However, it remains unclear how different conformations of the dimer contribute to the pore formation on the molecular level. Thus we have investigated various conformational states of the α9 dimer in a MOM model - using computer simulations supplemented with site-specific mutagenesis and crosslinking of the α9 helices. Our data not only confirmed the critical membrane environment for the α9 stability and dimerization, but also revealed the distinct lipid-binding preference of the dimer in different conformational states. In our proposed pathway, a crucial iso-parallel dimer that mediates the conformational transition was discovered computationally and validated experimentally. The corroborating evidence from simulations and experiments suggests that, helix α9 assists Bax activation via the dimer heterogeneity and interactions with specific MOM lipids, which eventually facilitate proteolipidic pore formation in apoptosis regulation. PMID:27381287

  13. Overexpression of Bax induces apoptosis and enhances drug sensitivity of hepatocellular cancer-9204 cells

    Institute of Scientific and Technical Information of China (English)

    Jian-Yong Zheng; Guang-Shun Yang; Wei-Zhong Wang; Jiang Li; Kai-Zong Li; Wen-Xian Guan; Wen-Liang Wang

    2005-01-01

    AIM: To investigate the role of overexpression of Bax in apoptotic pathways and the response of human hepatocellular cancer (HCC)-9204 cells to cell death induced by adriamycin.METHODS: The whole length of Bax cDNA was transfectrd into human HCC-9204 cells by the method of lipofectamine transfection. An inducible MT-Ⅱ regulatory system was constructed, which allowed controlled expression of protein upon addition of ZnSO4 (100 μmol/L) as an external inducer. Stable transfecting inducible expression vector containing Bax gene was performed. Expression of Bax in protein was analyzed by immunohistochemistry and Western blotting. TUNEL and flow cytometry were used to assess the effect of Bax on apoptosis. Colony assay and tetrazolium blue (MTT) assay were used to evaluate the difference in drug sensitivity of HCC-9204 cells after Bax-transfection.RESULTS: Immunohistochemistry and Western blotting demonstrated that the expression of Bax protein markedly increased in Bax-transfected cells 4 h after the addition cytoplasm and perinuclear region of HCC-9404 cells, and there was ectopic expression in cells with marked condensation of chromatin and cytoplasm (apoptotic cells). Apoptotic index significantly increased in Bax-transfected HCC-9204/Bax cells (3.6 vs 27.2, 4.2 vs 32.3, P<0.05).Flow cytometry analysis showed a significant sub-G1 peak and apoptosis in 15.4% HCC-9204/Bax cells 24 h after treatment. Furthermore, colony survival rate decreased from 66% (HCC-9204/pMD) to 45% (HCC-9204/Bax) 2 dafter ADR withdrawal. MTT assay result showed that the effects of Bax on cell viability following ADR exposure were significant as compared to the vehicle-transfected HCC-9204/pMD cells (21% vs 44%, P<0.01).CONCLUSION: Overexpression of Bax not only induces apoptosis, but also sensitizes HCC-9204 cells to cell death induced by adriamycin.

  14. Bax expression measured by AQUAnalysis is an independent prognostic marker in oral squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Bose Pinaki

    2012-08-01

    Full Text Available Abstract Background Resistance to apoptosis is a hallmark of cancer and proteins regulating apoptosis have been proposed as prognostic markers in several malignancies. However, the prognostic impact of apoptotic markers has not been consistently demonstrated in oral squamous cell carcinoma (OSCC. This inconsistency in reported associations between apoptotic proteins and prognosis can be partly attributed to the intrinsic low resolution and misclassification associated with manual, semi-quantitative methods of biomarker expression measurement. The aim of this study was to examine the association between apoptosis-regulating proteins and clinical outcomes in oral squamous cell carcinoma (OSCC using the quantitative fluorescence immunohistochemistry (IHC based AQUAnalysis technique. Methods Sixty-nine OSCC patients diagnosed between 1998–2005 in Calgary, Alberta, Canada were included in the study. Clinical data were obtained from the Alberta Cancer Registry and chart review. Tissue microarrays (TMAs were assembled from triplicate cores of formalin-fixed paraffin embedded pre-treatment tumour tissue. Bax, Bcl-2 and Bcl-XL protein expression was quantified using fluorescent IHC and AQUA technology in normal oral cavity squamous epithelium (OCSE and OSCC tumour samples. Survival was analyzed using Kaplan-Meier plots and the Cox proportional hazard model. Results Bax expression was predominantly nuclear in OCSE and almost exclusively cytoplasmic in OSCC. No similar differences in localization were observed for Bcl-2 or Bcl-XL. Only Bax expression associated with disease-specific survival (DSS, with 5-year survival estimates of 85.7% for high Bax versus 50.3% for low Bax (p = 0.006, in univariate analysis. High Bax expression was also significantly associated with elevated Ki67 expression, indicating that increased proliferation might lead to an improved response to radiotherapy in patients with elevated Bax expression. In multivariate analyses

  15. Bax expression measured by AQUAnalysis is an independent prognostic marker in oral squamous cell carcinoma

    International Nuclear Information System (INIS)

    Resistance to apoptosis is a hallmark of cancer and proteins regulating apoptosis have been proposed as prognostic markers in several malignancies. However, the prognostic impact of apoptotic markers has not been consistently demonstrated in oral squamous cell carcinoma (OSCC). This inconsistency in reported associations between apoptotic proteins and prognosis can be partly attributed to the intrinsic low resolution and misclassification associated with manual, semi-quantitative methods of biomarker expression measurement. The aim of this study was to examine the association between apoptosis-regulating proteins and clinical outcomes in oral squamous cell carcinoma (OSCC) using the quantitative fluorescence immunohistochemistry (IHC) based AQUAnalysis technique. Sixty-nine OSCC patients diagnosed between 1998–2005 in Calgary, Alberta, Canada were included in the study. Clinical data were obtained from the Alberta Cancer Registry and chart review. Tissue microarrays (TMAs) were assembled from triplicate cores of formalin-fixed paraffin embedded pre-treatment tumour tissue. Bax, Bcl-2 and Bcl-XL protein expression was quantified using fluorescent IHC and AQUA technology in normal oral cavity squamous epithelium (OCSE) and OSCC tumour samples. Survival was analyzed using Kaplan-Meier plots and the Cox proportional hazard model. Bax expression was predominantly nuclear in OCSE and almost exclusively cytoplasmic in OSCC. No similar differences in localization were observed for Bcl-2 or Bcl-XL. Only Bax expression associated with disease-specific survival (DSS), with 5-year survival estimates of 85.7% for high Bax versus 50.3% for low Bax (p = 0.006), in univariate analysis. High Bax expression was also significantly associated with elevated Ki67 expression, indicating that increased proliferation might lead to an improved response to radiotherapy in patients with elevated Bax expression. In multivariate analyses, Bax protein expression remained an independent

  16. bax, but not bcl-2, influences the prognosis of human pancreatic cancer

    OpenAIRE

    Friess, H; Lu, Z; H. Graber; Zimmermann, A.; Adler, G.; Korc, M.; Schmid, R; Buchler, M

    1998-01-01

    Background—bcl-2 and bax belong to the bcl-2-related gene family, which marks a new class of genes that influence apoptosis. The bcl-2 oncogene acts as a broad antiapoptotic factor and extends both normal and tumour cell survival. In contrast, the bax gene is a promoter of apoptosis. 
Aims—To analyse the expression of bcl-2 and bax in pancreatic cancer and correlate the results with clinical parameters. 
Patients—Pancreatic cancer tissue samples were obtained fro...

  17. Bcl-2 and bax expression and prostate cancer outcome in men treated with radiotherapy in Radiation Therapy Oncology Group protocol 86-10

    International Nuclear Information System (INIS)

    Purpose: Bcl-2 and bax are proteins with opposing roles in apoptosis regulation; yet abnormal expression of either has been associated with failure after radiotherapy (RT). In this study we examined bcl-2 and bax expression as predictive markers in men treated with radiotherapy ± androgen deprivation on Radiation Therapy Oncology Group (RTOG) protocol 86-10. Experimental Design: Suitable archival diagnostic tissue was obtained from 119 (26%) patients for bcl-2 analysis and 104 (23%) patients for bax analysis. Cox proportional hazards multivariate analysis was used to determine the relationship of abnormal bcl-2 and bax expression to the end points of local failure, distant metastasis, cause-specific mortality, and overall mortality. Bcl-2 overexpression was classified as any tumor cell cytoplasmic staining and altered bax expression was classified as greater or lesser cytoplasmic staining intensity of tumor cells as compared with adjacent normal prostate epithelium. Results: The study cohort exhibited bcl-2 overexpression in 26% (n = 30) of cases and abnormal bax expression in 47% (n = 49) of cases. A borderline significant relationship was observed between abnormal bax expression and higher Gleason score (p = 0.08). In univariate and multivariate analyses, there was no statistically significant relationship seen between abnormal bcl-2 or bax expression and outcome. Conclusions: Abnormal bcl-2 and bax expression were not related to any of the end points tested. The cohort examined was comprised of patients with locally advanced disease and it is possible that these markers may be of greater value in men with earlier-stage prostate cancer

  18. CEPO调控新生大鼠缺氧缺血性脑损伤Bcl-2、Bax基因表达的研究%Effect of CEPO on the expression of Bcl-2 and Bax genes in hypoxic ischemic brains of neonatal

    Institute of Scientific and Technical Information of China (English)

    付中秋; 姚笠; 田执梁; 张玉晶; 邵庆亮; 张海涛; 赵霞霞

    2014-01-01

    Objective To investigated the association between the expressions of Bcl-2 and Bax genes in neonatal rats upon cerebral hypoxic-ischemia and the effects of Carbamylated Erythropoietin (CEPO.) administration on both genes.Methods An animal model of neonatal hypoxic-ischemia brain damage was established.The changes of Bcl-2 and Bax mRNAs in brain tissues were detected in rats after HI or CEPO administration by RT-PCR.Results The levels of Bcl-2 and Bax mRNAs were increased upon HIBD compared with sham-operated group at 2 h,12 h,24 h,48 h,72 h time points post treatment (P < 0.05).On the contrary,only the level of Bcl-2 mRNA was increased (P < 0.05) while the level of Bax mRNA was decreased (P < 0.05) in CEPOintervention group.Conclusion The levels of Bcl-2 and Bax mRNAs in HIBD of neonatal rats were altered.CEPO may exert some neuroprotective effects on the brain tissues via interventing the expression of Bcl-2 and Bax.%目的 观察新生大鼠缺氧缺血性脑损伤(HIBD)脑组织Bcl-2、Bax基因表达变化及氨基甲酰化促红细胞生成素(CEPO)干预对其表达的影响,探讨CEPO发挥脑保护作用的可能机制.方法 将新生7日龄SD大鼠建立HIBD模型,利用RT-PCR检测缺氧缺血和CEPO干预后2h、12h、24 h、48 h、72 h凋亡基因Bcl-2、Bax的mRNA表达的改变.结果 与假手术组相比,缺氧缺组在2h、12h、24 h、48 h、72 h脑组织中Bcl-2、Bax的表达均增加(P<0.05),与缺氧缺血组相比,CEPO干预组在不同时间点Bcl-2表达增加(P<0.05),Bax表达下降(P<0.05).结论 在新生大鼠HIBD中Bcl-2、Bax mRNA的表达发生改变,调控二者的表达水平可能是CEPO发挥脑保护的作用机制之一.

  19. Apoptosis regulates ipRGC spacing necessary for rods and cones to drive circadian photoentrainment

    OpenAIRE

    Chen, Shih-Kuo; Chew, Kylie S.; McNeill, David S.; Keeley, Patrick W.; Ecker, Jennifer L.; Mao, Buqing Q.; Pahlberg, Johan; Kim, Bright; Lee, Sammy C. S.; Fox, Michael; Guido, William; Wong, Kwoon Y.; Sampath, Alapakkam P.; Reese, Benjamin E.; Kuruvilla, Rejji

    2013-01-01

    The retina consists of ordered arrays of individual types of neurons for processing vision. Here we show that such order is necessary for intrinsically photosensitive retinal ganglion cells (ipRGCs) to function as irradiance detectors. We found that during development, ipRGCs undergo proximity-dependent Bax-mediated apoptosis. Bax mutant mice exhibit disrupted ipRGC spacing and dendritic stratification with an increase in abnormally localized synapses. ipRGCs are the sole conduit for light in...

  20. Effect of Helicobacter pylori infection on Bax protein expression in patients with gastric precancerous lesions

    Institute of Scientific and Technical Information of China (English)

    Hai-Feng Liu; Wei-Wen Liu; Guo-An Wang; Xiao-Chun Teng

    2005-01-01

    AIM: To investigate the effect of Helicobacter pylori (H pylori) infection on Bax protein expression, and explore the role of H pylori in gastric carcinogenesis.METHODS: H pylori was assessed by rapid urease test and Warthin-Starry method, and expression of Bax protein was examined immunohistochemically in 72 patients with pre-malignant lesions.RESULTS: Bax protein was differently expressed in intestinal metaplasia and gastric dysplasia, and showed 63.99% positivity. The positivity of Bax protein expression in H pylori-positive gastric precancerous lesions (72.3%) was significantly higher than that in H pylori-negative gastric precancerous lesions (48.0%, χ2 = 4.191, P<0.05).H pylori infection was well correlated with the expression of Bax protein in gastric precancerous lesions (r = 0.978,P<0.01). After eradication of H pylori, the positivity of Bax protein expression significantly decreased in H pylori-positive gastric precancerous lesions (χ2= 5.506,P<0.05). In the persisting H pylori-infected patients,the positivity of Bax protein expression was not changed.CONCLUSION: H pylori infection may be involved in the upregulation of Bax gene, which might be one of the mechanisms of H pylori infection-induced gastric epithelial cell apoptosis. H pylori might act as a tumor promoter in the genesis of gastric carcinoma and eradication of H pylori could inhibit gastric carcinogenesis.

  1. Driving p53 Response to Bax Activation Greatly Enhances Sensitivity to Taxol by Inducing Massive Apoptosis

    Directory of Open Access Journals (Sweden)

    Paola De Feudis

    2000-05-01

    Full Text Available The proapoptotic gene bax is one of the downstream effectors of p53. The p53 binding site in the bax promoter is less responsive to p53 than the one in the growth arrest mediating gene p21. We introduced the bax gene under the control of 13 copies of a strong p53 responsive element into two ovarian cancer cell lines. The clones expressing bax under the control of p53 obtained from the wild-type (wt p53-expressing cell line A2780 were much more sensitive (500- to 1000-fold to the anticancer agent taxol than the parent cell line, with a higher percentage of cells undergoing apoptosis after drug treatment that was clearly p53-dependent and bax-mediated. Xenografts established in nude mice from one selected clone (A2780/C3 were more responsive to taxol than the parental line and the apoptotic response of A2780/C3 tumors was also increased after treatment. Introduction of the same plasmid into the p53 null SKOV3 cell line did not alter the sensitivity to taxol or the induction of apoptosis. In conclusion, driving the p53 response (after taxol treatment by activating the bax gene rather than the p21 gene results in induction of massive apoptosis, in vitro and in vivo, and greatly enhances sensitivity to the drug.

  2. The effect of radiation on bcl-2 and bax in hyperplastic prostatic tissues

    International Nuclear Information System (INIS)

    Aim: To investigate the expressions of bcl-2 and bax in benign prostatic hyperplasia (BPH) and the effect of β-rays on bcl-2 and bax. Methods: The expressions of bcl-2 and bax are studied by means of immunohistochemical method in 9 normal prostate (NP) and 15 BPH and 35 patients treated with 90Sr/90Y Prostatic Hyperplasia Applicator. Results: The expressions of bcl-2 in epithelia of NP and BPH are higher than that in stroma P<0.01=. The expressions of bcl-2 in epithelia and stroma of BPH are higher than that in NP P<0.01=. The expressions of bax in epithelia of NP are higher than that in BPH P<0.05=. However ,the expressions of bcl-2 in epithelia and stroma of BPH are higher than bax P<0.01 =. Compared with the control group, the expressions of bcl-2 in epithelia and stroma of BPH treated with 90Sr/90Y Prostatic Hyperplasia Applicator decreased and the expressions of bax increased P<0.01=. Conclusion: bcl-2 gene and bax gene play an important role in the regulation of prostatic apoptosis and the treatment of β-rays can accelerate the apoptosis of prostatic tissues. (authors)

  3. Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells

    Institute of Scientific and Technical Information of China (English)

    XU MaoJun; DONG JuFang

    2007-01-01

    Bax, a mammalian pro-apoptotic member of the Bcl-2 family, triggers hypersensitive reactions when expressed in plants. To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells, we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the β-estradiol-inducible promoter. The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on β-estradiol concentrations applied. Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 μmol/L β-estradiol are 5.0- and 5.5-fold of the control cells. Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str, two key genes in terpenoid indole alkaloid biosynthetic pathway of Catharanthus roseus cells, and stimulates the accumulation of defense-related protein PR1 in the cells, showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway. Thus, our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.

  4. Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Bax,a mammalian pro-apoptotic member of the Bcl-2 family,triggers hypersensitive reactions when expressed in plants.To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells,we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the β-estradiol-inducible promoter.The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on β-estradiol concentrations applied.Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 μmol/L β-estradiol are 5.0-and 5.5-fold of the control cells.Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str,two key genes in terpenoid indole alkaloid bio-synthetic pathway of Catharanthus roseus cells,and stimulates the accumulation of defense-related protein PR1 in the cells,showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway.Thus,our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.

  5. THE EFFECT OF EXERCISE PRECONDITION INDUCED MIR-21 AND BAX GENE EXPRESSIONS ON RATS' CARDIAC MYOCYTE AFTER EXHAUSTIVE EXERCISES%运动预适应对力竭运动后心肌mir-21和bax基因表达的研究

    Institute of Scientific and Technical Information of China (English)

    黄雅雯

    2011-01-01

    [目的]探讨运动预适应(excerise preconditioning,EP)对大鼠一次性力竭运动后心肌mir-21和bax基因表达的影响.[方法]健康雄性SD大鼠36只,随机分为对照组(C组)、一次性力竭运动组(E组)、运动预适应+一次性力竭运动组(EP组).力竭运动后即刻和24 h取各组心肌采用real-time PCR检测各心肌内mir-21和bax基因表达情况.[结果]EP可以显著抑制一次性力竭运动后心肌内mir-21基因的下调表达(P<0.05)和bax基因的上调表达(P<0.05).[结论]EP可以通过抑制mir-21基因下调从而抑制bax基因的上调来发挥保护作用.%[ Objective] To explore the effect of exercise preconditioning induced mir-21 and bax gene expressions on rats' cardiac myocyte after exhaustive exercises. [Methods] 36 healthy male SD rats were randomly divided into control group (C group) , exhaustive exercises (E group) , exercise preconditioning+ exhaustive exercises (EP group) .Used real-time PCR to detect expressions of mir-21 and bax after exhaustive exercises and 24h. [Results] The results of real-time PCR suggested that in EP group, down-regulation of mir-21 and up-regulation of bax cardiac myocyte were obviously inhibited by exhaustive exercises. [Conclusion] EP could inhibit the down-regulation of bax gene through inhibiting up-regulation of mir-21 gene.

  6. Bax/Bak activation in the absence of Bid, Bim, Puma, and p53.

    Science.gov (United States)

    Zhang, J; Huang, K; O'Neill, K L; Pang, X; Luo, X

    2016-01-01

    How BH3-only proteins activate Bax/Bak, the two gateway proteins of the mitochondria-dependent apoptotic pathway, remains incompletely understood. Although all pro-apoptotic BH3-only proteins are known to bind/neutralize the anti-apoptotic Bcl-2 proteins, the three most potent ones, Bid (tBid), Bim, and Puma, possess an additional activity of directly activating Bax/Bak in vitro. This latter activity has been proposed to be responsible for triggering Bax/Bak activation following apoptotic stimulation. To test this hypothesis, we generated Bid(-/)(-)Bim(-/)(-)Puma(-/)(-) (TKO), TKO/Bax(-/)(-)/Bak(-/)(-) (PentaKO), and PentaKO/Mcl-1(-/-) (HexaKO) HCT116 cells through gene editing. Surprisingly, although the TKO cells were resistant to several apoptotic stimuli, robust apoptosis was induced upon the simultaneous inactivation of Bcl-xL and Mcl-1, two anti-apoptotic Bcl-2 proteins known to suppress Bax/Bak activation and activity. Importantly, such apoptotic activity was completely abolished in the PentaKO cells. In addition, ABT-737, a BH3 mimetic that inhibits Bcl-xL/Bcl-w/Bcl-2, induced Bax activation in HexaKO cells reconstituted with endogenous level of GFP-Bax. Further, by generating TKO/p53(-/-) (QKO) cells, we demonstrated that p53, a tumor suppressor postulated to directly activate Bax, is not required for Bid/Bim/Puma-independent Bax/Bak activation. Together, these results strongly suggest that the direct activation activities of Bid (tBid), Bim, Puma, and p53 are not essential for activating Bax/Bak once the anti-apoptotic Bcl-2 proteins are neutralized. PMID:27310874

  7. Induction of Apoptosis and expression of Apoptosis-related gene products in response to radiation in murine tumors

    International Nuclear Information System (INIS)

    To analyze the involvement of apoptosis regulatory genes p53, p21waf1/cip1, bax and bcl-2 in induction of apoptosis by radiation in murine tumors. The radiation-sensitive ovarian carcinoma OCa-I and the radiation-resistant hepatocarcinoma HCa-I were used. Tumors, 8mm in diameter, were irradiated with 25Gy and at various times after irradiation, ranging from 1 to 48 h, were analyzed histologically for apoptosis and by western blot for alterations in the expression of these genes. The p53 status of the tumors were determined by the polymerase chain reaction-single strand conformation polymorphism assay. Both tumors were positive for wild-type p53. Radiation induced apoptosis in OCa-I but not in HCa-I. Apoptosis developed rapidly, peaked at 2 h after irradiation and returned to almost the background level at 48 h. In OCa-I radiation upregulated the expression of p53, p21waf1/cip1, and the bcl-2/bax ratio was decreased. In HCa-I radiation increased the expression of both p53 and p21waf1/cip1, although the increase of the latter was small. The bcl-2/bax ratio was greatly increased. In general the observed changes occurred within a few hours after irradiation, and either preceded or coincided with development of apoptosis. The development of apoptosis required upregulation of both p53 and p21waf1/cip1 as well as a decrease in bcl-2/bax ratio. In contrast, an increase in bcl-2/bax ratio prevented apoptosis in the presence of upregulated p53 and p21waf1/cip1. These findings identified the involvement of multiple oncogenes in apoptosis regulation in vivo and demonstrate the complexity that may be associated with the use of a single oncogene assessment for predicting the outcome of cancer therapy with cytotoxic agents. (author)

  8. GSH depletion enhances adenoviral bax-induced apoptosis in lung cancer cells.

    Science.gov (United States)

    Honda, Tsuyoshi; Coppola, Simona; Ghibelli, Lina; Cho, Song H; Kagawa, Shunsuke; Spurgers, Kevin B; Brisbay, Shawn M; Roth, Jack A; Meyn, Raymond E; Fang, Bingliang; McDonnell, Timothy J

    2004-04-01

    The utility of dominant acting proapoptotic molecules to induce cell death in cancer cells is being evaluated in preclinical studies and clinical trials. We recently developed a binary adenoviral expression system to enable the efficient gene transfer of Bax and other proapoptotic molecules. Using this system, overexpression of Bax protein in four non-small-cell lung cancer (NSCLC) cell lines, H1299, A549, H226 and H322, was evaluated. The H322 line exhibited significant resistance to Bax-induced cell death compared to the other cell lines. H322 cells had the highest level of glutathione (GSH). GSH levels were significantly decreased following buthionine sulfoximine treatment and this coincided with enhanced apoptosis induction by Ad-Bax in H322 cells. GSH depletion enhanced Bax protein translocation to mitochondrial membranes. These findings suggest that the redox status may be a determinant of Bax-mediated cell death and that manipulation of intracellular thiols may sensitize cells to apoptosis by facilitating Bax insertion into mitochondrial membranes. PMID:15002033

  9. A Novel Mechanism for CTCF in the Epigenetic Regulation of Bax in Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Claudia Fabiola Méndez-Catalá

    2013-08-01

    Full Text Available We previously reported the association of elevated levels of the multifunctional transcription factor, CCCTC binding factor (CTCF, in breast cancer cells with the specific anti-apoptotic function of CTCF. To understand the molecular mechanisms of this phenomenon, we investigated regulation of the human Bax gene by CTCF in breast and non-breast cells. Two CTCF binding sites (CTSs within the Bax promoter were identified. In all cells, breast and non-breast, active histone modifications were present at these CTSs, DNA harboring this region was unmethylated, and levels of Bax mRNA and protein were similar. Nevertheless, up-regulation of Bax mRNA and protein and apoptotic cell death were observed only in breast cancer cells depleted of CTCF. We proposed that increased CTCF binding to the Bax promoter in breast cancer cells, by comparison with non-breast cells, may be mechanistically linked to the specific apoptotic phenotype in CTCF-depleted breast cancer cells. In this study, we show that CTCF binding was enriched at the Bax CTSs in breast cancer cells and tumors; in contrast, binding of other transcription factors (SP1, WT1, EGR1, and c-Myc was generally increased in non-breast cells and normal breast tissues. Our findings suggest a novel mechanism for CTCF in the epigenetic regulation of Bax in breast cancer cells, whereby elevated levels of CTCF support preferential binding of CTCF to the Bax CTSs. In this context, CTCF functions as a transcriptional repressor counteracting influences of positive regulatory factors; depletion of breast cancer cells from CTCF therefore results in the activation of Bax and apoptosis.

  10. Antitumor Activities and Apoptosis-regulated Mechanisms of Fermented Wheat Germ Extract in the Transplantation Tumor Model of Human HT-29 Cells in Nude Mice

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jia Yan; XIAO Xiang; DONG Ying; WU Jing; ZHOU Xing Hua

    2015-01-01

    Objective A subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermented wheat germ with Lactobacillus plantarum dy-1 (LFWGE). Methods The HT-29 cells were transplanted via subcutaneous injection of 1×107 cells into the right flank of each nude mouse. Then, nude mice were treated for 30 d with LFWGE (high-dose 2 g/kg/d;low-dose 1 g/kg/d) and for 7 d with 5-fluorouracil (5-FU, 25 mg/kg/d) by gavage and intraperitoneal injection, respectively. An inhibition of tumor growth was observed. Results Tumor volume and weights decreased significantly in both groups of nude mice treated with LFWGE. In addition, the cell apoptosis rate of the LFWGE group (2 g/kg/d, 60.1%±4.4%; 1 g/kg/d, 58.6%±6.9%) was significantly higher than that of the control group (11.5%±1.6%) and 5-FU group (32.1%±3.5%) as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot method further confirmed these enhancing apoptosis and growth inhibition effects. The involvement of LFWGE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, Caspase-3, and CyclinD1. Conclusion The results showed that LFWGE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be as a natural nutrient supplements or chemopreventive agent in the treatment of human colon cancer.

  11. Effect of β Radiation on Bcl-2 and Bax Expressions in Benign Prostate Hyperplasia Tissues

    Institute of Scientific and Technical Information of China (English)

    MA Qing-jie; GAO Shi; ZHAO Jie; GU Xin-quan; CAI Shan-yu; ZHAO Guo-qing

    2008-01-01

    The authors chose specimens from nine normal prostate tissues(NP group),15 benign prostate hyperplasia(BPH) prostates(BPH group),and 35 BPH prostates that had been treated with 90Sr/90Y Prostatic Hyperplasia Applicator(exposure group),The expressions of bcl-2 and bax in stroma and epithelia of prostate tissues were demonstrated by means of immunohistochemical staining,and the staining positive rate was semiquantatively determined,so as to observe the expression of bcl-2 and bax genes in the prostate tissues of normal individuals and BPH patients,before and after β radiation,and to evaluate the influence of β radiation on bcl-2 and bax expressions,The expressions of gene bcl-2 in the prostate epithelia of NP and BPH are significantly higher than those in the prostate stroma(P<0.01),However,the expressions of bcl-2 in the prostate epithelia and stroma of the BPH group are obviously higher than those in the NP group(P<0.01),The expression of gene bax in the prostate epithelia of the NP group is higher than that in the BPH group(P<0.05),However,bcl-2 expressions in the prostate epithelia and stroma of the BPH group are significantly higher than the bax expressions(P<0.01),Compared with those of the NP group,the expressions of bcl-2 in the prostate epithelia and stroma of the exposure group decrease remarkably,even as the expressions of the bax notably increase(P<0.01),Thus,the administration of β radiation can remarkably affect bcl-2 and bax gene expressions,to regulate cell apoptosis,in the prostate tissues of BPH.

  12. Impact of resveratrol on the expression of apoptosis related gene survivin and bax in human cancer cells%白藜芦醇对食管癌细胞凋亡相关基因survivin和bax表达的影响

    Institute of Scientific and Technical Information of China (English)

    Yongjun Li; Xiaohui Sun; Rui Zhang

    2009-01-01

    Objective: We explored the mechanism of apoptosis in human esophageal cancer Eca109 cells by resveratrol.Methods: The suppressive ratio of resveratrol on Ecal09 cells proliferation was evaluated by MTT colorimetric assay and morphology was observed by transmission electron microscope. The expression of survivin and bax was analyzed by RT-PCR and Flow Cytometry (FCM). Results: Resveratrel inhibited the growth of Ecal09 cells in a dose-and time-dependent man-ner, and the suppressive ratio arrived at 76.42%. Morphological apoptosis could be observed after treated with resveratrol.The bulk of some drug-treated cells turned small and the nuclear chromatin became condensed and marginated. The results determined by RT-PCR and FCM showed that resveratrol could down-regulate surviving, while up-regulate bax. Conclusion:Resveratrol could induce the apoptosis of human esophageal cancer Eca109 cells, and its possible molecular mechanisms might be related to modulation the expression of survivin and bax.

  13. Study on the Relationship between the Gene Expression of Bax and Apoptosis in Tongue Mycoderma Epithelium of Common Tongue Coating%bax mRNA和蛋白产物与常见舌苔舌上皮细胞凋亡关系的研究靠

    Institute of Scientific and Technical Information of China (English)

    李明; 吴正治; 何朝; 张永锋; 陈嫚茵

    2003-01-01

    目的:检测常见舌苔舌上皮细胞凋亡情况及凋亡相关基因bax mRNA和蛋白产物,探讨舌苔厚度变化与舌上皮细胞凋亡、bax基因表达的关系.方法:运用TUNEL(末端脱氧核苷酸转移酶介导的脱氧尿嘧啶核苷三磷酸缺口末端标记)技术、原位杂交、免疫组化和图像分析技术.结果:与正常薄苔比较,剥苔bax基因过度表达伴随细胞凋亡增多,而厚苔bax基因低表达伴随细胞凋亡减少.bax基因表达水平变化趋势与细胞凋亡水平变化趋势一致.结论:bax基因表达水平的变化可能是影响舌苔上皮细胞凋亡并导致舌苔厚度变化的重要原因.

  14. Immunohistochemical localization of Bcl-2 and Bax proteins in in situ and invasive duct breast carcinomas.

    Science.gov (United States)

    Kapucuoglu, N; Losi, L; Eusebi, V

    1997-01-01

    Bcl-2 and Bax proteins are coded by a family of genes that take part in the manteinance of the balance between cell proliferation rate and programmed cell death in multicellular organisms. The Bax gene acts as promoter of cell death by opposing the death protector effect of the Bcl-2 gene. Expression of the Bcl-2 and Bax proteins has been investigated in 58 cases of duct carcinoma in situ (DCIS) and duct invasive and invasive lobular carcinomas (IC) of the breast. While both proteins were expressed at the same time in normal and benign epithelium, different staining patterns were observed according to the degree of differentiation of the neoplastic epithelium. In well-differentiated DCIS and grade I IC there was a predominance of Bcl-2 protein staining. Grade II lesions co-expressed both proteins. Poorly differentiated DCIS displayed a predominantly Bax protein staining pattern. Therefore, it appears that Bax protein expression, especially in DCIS, relates to more aggressive neoplasms while Bcl-2 protein expression is associated with less aggressive malignant lesions.

  15. Influence of Apoptin on Up-regulation of the Expression of Bad and Bax

    Institute of Scientific and Technical Information of China (English)

    GUO Tai; YANG Qian

    2005-01-01

    The chicken anemia virus protein, apoptin, which manifests selectivity and specificity to tumor cells, induces a p53-independent and Bcl-2-insensitive type of apoptosis in various human tumor cells. In this study, the apoptin gene was cloned from the total DNA of chicken anemia virus, and the recombinant vector was constructed. We used oligonucleotide microarray to study the changes of four genes, including Bcl-2, Bcl-xL, Bad and Bax. The post-transfection with the recombinant was also studied. The pro-apoptotic genes(Bad and Bax) and anti-apoptosis genes(Bcl-2 and Bcl-xL) were up-regulated in contrast to the controls. According to the published data, either Bcl-2 or Bcl-xL can form non-functional heterodimers by Bad and Bax binding together, resulting in blocking partly the release of cytochrome c from mitochondria. However, apoptosis could be inhibited by neither the endogenous Bcl-xL nor Bcl-2 over-expression. The experiments show that the apoptin-induced apoptotic pathway is related to the up-regulation of Bad and Bax. Bad was up-regulated by apoptin; then this up-regulated product of Bad was in favor of displacing Bax from binding to Bcl-xL or Bcl-2. Consequently, Bax exerted a pro-apoptotic dysfunction to mitochondria, thereby inducing the release of cytochrome c. Finally, apoptin induced the apoptosis of HHCC cells. These results indicate that the oligonucleotide microarray can reveal the genes related to the apoptosis induced by apoptin in HHCC cells.

  16. Proteomics analysis of apoptosis-regulating proteins in tissues with different radiosensitivity

    International Nuclear Information System (INIS)

    The aim of this study was to identify of radiosusceptibility proteins in tissues with different radiosensitivity. C3H/HeJ mice were exposed to 10 Gy. The tissues were processed for proteins extraction and were analyzed by 2-dimensional electrophoresis. The proteins were identified by matrix-assisted laser desorption ionizing time-of-flight mass spectrometry and validated by immunohistochemical staining and Western blotting. The peaks of apoptosis levels were 35.3±1.7% and 0.6±0.2% in the spleen and the liver, respectively, after ionizing radiation. Analysis of liver tissue showed that the expression level of reactive oxygen species (ROS) related proteins such as cytochrome c, glutathione S transferase, NADH dehydrogenase and peroxiredoxin VI increased after radiation. The expression level of cytochrome c increased to 3-fold after ionizing radiation in both tissues. However in spleen tissue, the expression level of various kinds of apoptosis regulating proteins increased after radiation. These involved iodothyronine, CD 59A glycoprotein precursor, fas antigen and tumor necrosis factor -inducible protein TSG-6nprecursor after radiation. The difference in the apoptosis index between the liver and spleen tissues is closely associated with the expression of various kinds of apoptosis-related proteins. The result suggests that the expression of apoptosis-related protein and redox proteins play important roles in this radiosusceptibility. (author)

  17. The X-linked inhibitor of apoptosis regulates long-term depression and learning rate.

    Science.gov (United States)

    Gibon, Julien; Unsain, Nicolas; Gamache, Karine; Thomas, Rhalena A; De Leon, Andres; Johnstone, Aaron; Nader, Karim; Séguéla, Philippe; Barker, Philip A

    2016-09-01

    Hippocampal long-term depression (LTD) is an active form of synaptic plasticity that is necessary for consolidation of spatial memory, contextual fear memory, and novelty acquisition. Recent studies have shown that caspases (CASPs) play an important role in NMDA receptor-dependent LTD and are involved in postsynaptic remodeling and synaptic maturation. In the present study, we examined the role of X-linked inhibitor of apoptosis (XIAP), a putative endogenous CASP inhibitor, in synaptic plasticity in the hippocampus. Analysis in acute brain slices and in cultured hippocampal neurons revealed that XIAP deletion increases CASP-3 activity, enhances α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor internalization, sharply increases LTD, and significantly reduces synapse density. In vivo behaviors related to memory were also altered in XIAP(-/-) mice, with faster acquisition of spatial object location and increased fear memory observed. Together, these results indicate that XIAP plays an important physiologic role in regulating sublethal CASP-3 activity within central neurons and thereby facilitates synaptic plasticity and memory acquisition.-Gibon, J., Unsain, N., Gamache, K., Thomas, R. A., De Leon, A., Johnstone, A., Nader, K., Séguéla, P., Barker, P. A. The X-linked inhibitor of apoptosis regulates long-term depression and learning rate.

  18. BAX supports the mitochondrial network, promoting bioenergetics in nonapoptotic cells

    Science.gov (United States)

    Boohaker, Rebecca J.; Zhang, Ge; Carlson, Adina Loosley; Nemec, Kathleen N.

    2011-01-01

    The dual functionality of the tumor suppressor BAX is implied by the nonapoptotic functions of other members of the BCL-2 family. To explore this, mitochondrial metabolism was examined in BAX-deficient HCT-116 cells as well as primary hepatocytes from BAX-deficient mice. Although mitochondrial density and mitochondrial DNA content were the same in BAX-containing and BAX-deficient cells, MitoTracker staining patterns differed, suggesting the existence of BAX-dependent functional differences in mitochondrial physiology. Oxygen consumption and cellular ATP levels were reduced in BAX-deficient cells, while glycolysis was increased. These results suggested that cells lacking BAX have a deficiency in the ability to generate ATP through cellular respiration. This conclusion was supported by detection of reduced citrate synthase activity in BAX-deficient cells. In nonapoptotic cells, a portion of BAX associated with mitochondria and a sequestered, protease-resistant form was detected. Inhibition of BAX with small interfering RNAs reduced intracellular ATP content in BAX-containing cells. Expression of either full-length or COOH-terminal-truncated BAX in BAX-deficient cells rescued ATP synthesis and oxygen consumption and reduced glycolytic activity, suggesting that this metabolic function of BAX was not dependent upon its COOH-terminal helix. Expression of BCL-2 in BAX-containing cells resulted in a subsequent loss of ATP measured, implying that, even under nonapoptotic conditions, an antagonistic interaction exists between the two proteins. These findings infer that a basal amount of BAX is necessary to maintain energy production via aerobic respiration. PMID:21289292

  19. BAX OVEREXPRESSION ENHANCES APOPTOSIS INDUCED BY ADRIAMYCIN IN HCC-9204 CELLS

    Institute of Scientific and Technical Information of China (English)

    郑建勇; 李江; 李开宗; 王文亮; 王为忠

    2004-01-01

    Objective: To investigate the effect of overexpression of Bax to the sensitivity of human HCC-9204 cells to adriamycin (ADR). Methods: Human cultured hepatocellular carcinoma cell line HCC-9204 was exposed in vitro to adriamycin for various time. An inducible vector containing Bax gene, with ZnSO4 as external inducer was constructed. Cell apoptosis was ascertained by morphological criteria, detection of apoptotic DNA fragmentation by TUNEL assay and flow cytometry. Tetrazolium blue (MTT) assay was used to evaluate the differences in drug sensitivity of HCC-9204 cells after Bax-transfection. Results: HCC-9204 cells treated with adriamycin at 20μmol/L showed extensive cell death. TUNEL assay showed nucleus fragmentation. And apoptotic peak was also shown by flow cytometry. FACS analyses showed a significant sub-G1 peak and apoptosis in 31% cells at 24h after treatment. Furthermore, the time-course of cell viability following exposure of HCC-9204/Bax cells to adriamycin showed that Bax was able to significantly decrease cell survival following exposure to adriamycin.

  20. Apoptosis repressor with a CARD domain (ARC restrains Bax-mediated pathogenesis in dystrophic skeletal muscle.

    Directory of Open Access Journals (Sweden)

    Jennifer Davis

    Full Text Available Myofiber wasting in muscular dystrophy has largely been ascribed to necrotic cell death, despite reports identifying apoptotic markers in dystrophic muscle. Here we set out to identify the contribution of canonical apoptotic pathways to skeletal muscle degeneration in muscular dystrophy by genetically deleting a known inhibitor of apoptosis, apoptosis repressor with a card domain (Arc, in dystrophic mouse models. Nol3 (Arc protein genetic deletion in the dystrophic Sgcd or Lama2 null backgrounds showed exacerbated skeletal muscle pathology with decreased muscle performance compared with single null dystrophic littermate controls. The enhanced severity of the dystrophic phenotype associated with Nol3 deletion was caspase independent but dependent on the mitochondria permeability transition pore (MPTP, as the inhibitor Debio-025 partially rescued skeletal muscle pathology in Nol3 (-/- Sgcd (-/- double targeted mice. Mechanistically, Nol3 (-/- Sgcd (-/- mice showed elevated total and mitochondrial Bax protein levels, as well as greater mitochondrial swelling, suggesting that Arc normally restrains the cell death effects of Bax in skeletal muscle. Indeed, knockdown of Arc in mouse embryonic fibroblasts caused an increased sensitivity to cell death that was fully blocked in Bax Bak1 (genes encoding Bax and Bak double null fibroblasts. Thus Arc deficiency in dystrophic muscle exacerbates disease pathogenesis due to a Bax-mediated sensitization of mitochondria-dependent death mechanisms.

  1. Ernest Belfort Bax: Marxist, Idealist, Positivist

    OpenAIRE

    Bevir, Mark

    1993-01-01

    Bax was the leading philosopher of the socialist revival in Britain during the 1880s. He saw Marxism as an economic and historical science that lacked a philosophical and ethical basis. Consequently, he tried to justify the Marxian dialectic by using a philosophy indebted to German idealism to show that the dialectic was a fact about reality itself, and he also tried to provide an ethical defence of Marxism in terms of a positivist ethic enshrining the goals of the French Revolution. Such ...

  2. Small molecules reveal an alternative mechanism of Bax activation.

    Science.gov (United States)

    Brahmbhatt, Hetal; Uehling, David; Al-Awar, Rima; Leber, Brian; Andrews, David

    2016-04-15

    The pro-apoptotic protein Bax commits a cell to death by permeabilizing the mitochondrial outer membrane (MOM). To obtain small-molecule probes for elucidating the molecular mechanism(s) of Bax activation, we screened for compounds that induced Bax-mediated liposome permeabilization. We identified five structurally different small molecules that promoted both Bax targeting to and oligomerization at membranes. All five compounds initiated Bax oligomerization in the absence of membranes by a mechanism unlike Bax activation by Bcl-2 homology 3 domain (BH3) proteins. Some of the compounds induced Bax/Bak-dependent apoptosis in cells. Activation of Bax by the most active compound was poorly inhibited by the anti-apoptotic protein Bcl-XL and requires a cysteine residue at position 126 of Bax that is not required for activation by BH3 proteins. Our results reveal a novel pathway for Bax activation independent of pro-apoptotic BH3 proteins that may have important implications for the regulation of Bax activity in cells. PMID:26916338

  3. 黄芪多糖对力竭运动大鼠心肌细胞凋亡及BcL-2、Bax蛋白表达的影响%Effects of acute exercise to exhaustion on BcL-2,Bax gene of rat, sheart after supplyment of astragalus polysaccharide

    Institute of Scientific and Technical Information of China (English)

    马兰军; 田振军

    2012-01-01

    为了探讨黄芪多糖(APS)对力竭运动大鼠机体的保护作用,通过建立大强度力竭运动大鼠模型,选取成年健康雄性SD大鼠24只,随机分为安静对照组,运动组和运动加药组,运动组和运动加药组按照训练模型进行为期6周的耐力训练,最后1次训练进行1次力竭运动,随后取心肌组织并进行样本处理.检测大鼠心肌MDA含量,用缺口末端标记法(TUNEL法)及免疫组织化学法检大鼠心肌细胞凋亡指数(AI)及BcL-2、Bax蛋白表达的变化.结果表明:运动组MDA含量、AI和BcL-2、Bax蛋白表达水平均明显高于安静对照组(P<0.05);与运动组比较,运动加药组MDA含量,AI与Bax蛋白表达水平均下降,而BcL-2蛋白表达水平显著增高(P<0.05).黄芪多糖可有效抑制力竭运动大鼠心肌细胞凋亡,此作用可能与其减轻氧化应激、上调BcL-2和下调Bax蛋白表达水平有关.%To study the supplementary astragalus polysaccharide (APS) to exhaustion movement rats myocardial cell apoptosis, observe the influence of myocardial cell apop-tosis BcL-2, the regulation of gene Bax changes, explore astragalus polysaccharide (APS) to exhaustion movement rats airframe of protection. By establishing high intensity exhaustion movement model of rats, the selection of adult health male SD rats 24 only randomly divided into quiet in the exercise group and control group, sports dosing group, the exercise group and exercise dosing groups according to training model six-week endurance training, last training once exhaustion movement, exhaustion after motion myocardial organize and sample processing. Detection rats myocardial MDA content, using gap end labeling (TUNEL law) and the method of immunohis to chemistry methods were used by rats myocardial cell apoptosis I,ndex (AI) and BcL-2, Bax protein expression changes. In the exercise group MDA content, AI and BcL-2, Bax protein expression levels were significantly higher in quiet in control group (P

  4. WNT signaling controls expression of pro-apoptotic BOK and BAX in intestinal cancer

    Energy Technology Data Exchange (ETDEWEB)

    Zeilstra, Jurrit; Joosten, Sander P.J. [Department of Pathology, Academic Medical Center, University of Amsterdam (Netherlands); Wensveen, Felix M. [Department of Experimental Immunology, Academic Medical Center, Amsterdam (Netherlands); Dessing, Mark C.; Schuetze, Denise M. [Department of Pathology, Academic Medical Center, University of Amsterdam (Netherlands); Eldering, Eric [Department of Experimental Immunology, Academic Medical Center, Amsterdam (Netherlands); Spaargaren, Marcel [Department of Pathology, Academic Medical Center, University of Amsterdam (Netherlands); Pals, Steven T., E-mail: s.t.pals@amc.uva.nl [Department of Pathology, Academic Medical Center, University of Amsterdam (Netherlands)

    2011-03-04

    Research highlights: {yields} Intestinal adenomas initiated by aberrant activation of the WNT pathway displayed an increased sensitivity to apoptosis. {yields} Expression profiling of apoptosis-related genes in Apc{sup Min/+} mice revealed the differential expression of pro-apoptotic Bok and Bax. {yields} APC-mutant adenomatous crypts in FAP patients showed strongly increased BAX immunoreactivity. {yields} Blocking of {beta}-catenin/TCF-4-mediated signaling in colon cancer cells reduced the expression of BOK and BAX. -- Abstract: In a majority of cases, colorectal cancer is initiated by aberrant activation of the WNT signaling pathway. Mutation of the genes encoding the WNT signaling components adenomatous polyposis coli or {beta}-catenin causes constitutively active {beta}-catenin/TCF-mediated transcription, driving the transformation of intestinal crypts to cancer precursor lesions, called dysplastic aberrant crypt foci. Deregulated apoptosis is a hallmark of adenomatous colon tissue. However, the contribution of WNT signaling to this process is not fully understood. We addressed this role by analyzing the rate of epithelial apoptosis in aberrant crypts and adenomas of the Apc{sup Min/+} mouse model. In comparison with normal crypts and adenomas, aberrant crypts displayed a dramatically increased rate of apoptotic cell death. Expression profiling of apoptosis-related genes along the crypt-villus axis and in Apc mutant adenomas revealed increased expression of two pro-apoptotic Bcl-2 family members in intestinal adenomas, Bok and Bax. Analysis of the colon of familial adenomatous polyposis (FAP) patients along the crypt-to-surface axis, and of dysplastic crypts, corroborated this expression pattern. Disruption of {beta}-catenin/TCF-4-mediated signaling in the colorectal cancer cell line Ls174T significantly decreased BOK and BAX expression, confirming WNT-dependent regulation in intestinal epithelial cells. Our results suggest a feedback mechanism by which

  5. WNT signaling controls expression of pro-apoptotic BOK and BAX in intestinal cancer

    International Nuclear Information System (INIS)

    Research highlights: → Intestinal adenomas initiated by aberrant activation of the WNT pathway displayed an increased sensitivity to apoptosis. → Expression profiling of apoptosis-related genes in ApcMin/+ mice revealed the differential expression of pro-apoptotic Bok and Bax. → APC-mutant adenomatous crypts in FAP patients showed strongly increased BAX immunoreactivity. → Blocking of β-catenin/TCF-4-mediated signaling in colon cancer cells reduced the expression of BOK and BAX. -- Abstract: In a majority of cases, colorectal cancer is initiated by aberrant activation of the WNT signaling pathway. Mutation of the genes encoding the WNT signaling components adenomatous polyposis coli or β-catenin causes constitutively active β-catenin/TCF-mediated transcription, driving the transformation of intestinal crypts to cancer precursor lesions, called dysplastic aberrant crypt foci. Deregulated apoptosis is a hallmark of adenomatous colon tissue. However, the contribution of WNT signaling to this process is not fully understood. We addressed this role by analyzing the rate of epithelial apoptosis in aberrant crypts and adenomas of the ApcMin/+ mouse model. In comparison with normal crypts and adenomas, aberrant crypts displayed a dramatically increased rate of apoptotic cell death. Expression profiling of apoptosis-related genes along the crypt-villus axis and in Apc mutant adenomas revealed increased expression of two pro-apoptotic Bcl-2 family members in intestinal adenomas, Bok and Bax. Analysis of the colon of familial adenomatous polyposis (FAP) patients along the crypt-to-surface axis, and of dysplastic crypts, corroborated this expression pattern. Disruption of β-catenin/TCF-4-mediated signaling in the colorectal cancer cell line Ls174T significantly decreased BOK and BAX expression, confirming WNT-dependent regulation in intestinal epithelial cells. Our results suggest a feedback mechanism by which uncontrolled epithelial cell proliferation in the stem

  6. Detect of P53,bax and caspase 3 genes expression after optic nerve injury in rats with SYBR green I fluorescence quantitative PCR%SYBR Green I荧光定量PCR检测大鼠外伤性视神经损伤后P53、bax和caspase 3基因表达

    Institute of Scientific and Technical Information of China (English)

    吕瀛娟; 赵秀兰; 杨洁; 于金国; 颜华

    2009-01-01

    目的 应用SYBR Green Ⅰ荧光定量PCR法检测外伤性视神经损伤后P53、bax和caspase 3基因mRNA表达的变化.方法 应用液压颅脑损伤仪建立大鼠外伤性视神经损伤动物模型,伤后1、3、5、7、9、14、28d处死,以Trizol法提取新鲜视网膜组织的总RNA,以Oligo (dt) 18 为引物逆转录合成cDNA 并进行扩增,以T/A克隆法将纯化的目的 片断与T/A克隆载体(pTZ57R/T)连接成重组质粒并转化入E.coli DH5α.采用碱裂解法提取重组质粒,经蓝白斑筛选、酶切、测序鉴定后,根据标准品建立标准曲线,由软件自动计算出待测样本中靶基因mRNA的含量,并以靶基因和内参GAPDH mRNA含量的比值作为评价靶基因表达水平的指标.结果 由pTZ57R/T与目的 基因所构建的标准曲线的线性关系良好、灵敏度高、特异性强、准确可靠.P53和bax均在视神经损伤后3d mRNA表达明显增加,5d时达到高峰,7d后开始下降;伤后5d caspase 3 mRNA表达明显增加,9d时达到高峰,14d后开始下降.三者表达水平与对照组相比,差异均有统计学意义(P<0.05).结论 促凋亡基因P53、bax和caspase 3在视神经损伤后视网膜神经节细胞(RGCs)的凋亡发生中起到重要作用.%Objective Previous study showed that the histopathological basis of visual function damage caused by optical nerve injury is apoptosis of retinal ganglion cells(RGCs). This procedure is regulated by P53, bax and caspase 3 genes. Present study aimed to observe the expression of bax, P53 and caspase 3 mRNA in RGCs after traumatic optic nerve damage in the rats by SYBR green I fluorescence quantitative PCR method. Methods The animal model of optic nerve injury was established in the right eyes of 56 adult Wistar rats by a fluid percussion brain injury device (FPI) . Animal were killed on days 1, 3, 5, 7, 9, 14, 28 days separately after injury. Other 16 Wistar rats were divided into normal control group and sham operation group. The total RNA

  7. Association of Bax Expression and Bcl2/Bax Ratio with Clinical and Molecular Prognostic Markers in Chronic Lymphocytic Leukemia

    Directory of Open Access Journals (Sweden)

    Vucicevic Ksenija

    2016-04-01

    Full Text Available Background: In chronic lymphocytic leukemia (CLL, in vivo apoptotic resistance of malignant B lymphocytes results, in part, from the intrinsic defects of their apoptotic machinery. These include genetic alterations and aberrant expression of many apoptosis regulators, among which the Bcl2 family members play a central role.

  8. Effect of Bcl-2 and Bax on survival of side population cells from hepatocellular carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To understand the role and significance of side population (SP) cells from hepatocellular carcinoma (HCC) in hepatocarcinogenesis, development, relapse and metastasis, we simulated the denutrition conditions that cancer cells experience in clinical therapy, observed the different anti-apoptosis ability of SP cells and non-SP cells under such conditions, and established the possible effects of P53, Bcl-2 and Bax on survival of SP cells.METHODS: We used flow cytometry to analyze and sort the SP and non-SP cells in established HCC lines MHCC97and hHCC. We evaluated cell proliferation by methyl thiazolyl tetrazolium (MTT) assay and investigated the expression of p53, bd-2 and bax genes during denutrition,by RT-PCR and immunofluorescence staining.RESULTS: The percentage of SP cells in the two established HCC lines was 0.25% and 0.5%, respectively.SP cells had greater anti-apoptosis and proliferation ability than non-SP cells. Expression of Bcl-2 and Bax in SP and non-SP cells differed during denutrition. The former was up-regulated in SP cells, and the latter was up-regulated in non-SP cells.CONCLUSION: It may be that different upstream molecules acted and led to different expression levels of Bcl-2 and Bax in these two cell lines. There was a direct relationship between up-regulation of Bcl-2 and down-regulation of Bax and higher anti-apoptosis ability in SP cells. It may be that the existence and activity of SP cells are partly responsible for some of the clinical phenomena which are seen in HCC, such as relapse or metastasis. Further research on SP cells may have potential applications in the field of anticancer therapy.

  9. Cell cycle and apoptosis regulation by NFAT transcription factors: new roles for an old player.

    Science.gov (United States)

    Mognol, G P; Carneiro, F R G; Robbs, B K; Faget, D V; Viola, J P B

    2016-01-01

    The NFAT (nuclear factor of activated T cells) family of transcription factors consists of four Ca(2+)-regulated members (NFAT1-NFAT4), which were first described in T lymphocytes. In addition to their well-documented role in T lymphocytes, where they control gene expression during cell activation and differentiation, NFAT proteins are also expressed in a wide range of cells and tissue types and regulate genes involved in cell cycle, apoptosis, angiogenesis and metastasis. The NFAT proteins share a highly conserved DNA-binding domain (DBD), which allows all NFAT members to bind to the same DNA sequence in enhancers or promoter regions. The same DNA-binding specificity suggests redundant roles for the NFAT proteins, which is true during the regulation of some genes such as IL-2 and p21. However, it has become increasingly clear that different NFAT proteins and even isoforms can have unique functions. In this review, we address the possible reasons for these distinct roles, particularly regarding N- and C-terminal transactivation regions (TADs) and the partner proteins that interact with these TADs. We also discuss the genes regulated by NFAT during cell cycle regulation and apoptosis and the role of NFAT during tumorigenesis. PMID:27100893

  10. PUMA and Bax-induced Autophagy Contributes to Apoptosis

    OpenAIRE

    Yee, Karen S.; Wilkinson, Simon; James, John; Ryan, Kevin M.; Vousden, Karen H.

    2009-01-01

    The p53-inducible BH3-only protein PUMA is a key mediator of p53-dependent apoptosis, and PUMA has been shown to function by activating Bax and mitochondrial outer membrane permeabilization. In this study we describe an ability of PUMA to induce autophagy that leads to the selective removal of mitochondria. This function of PUMA depends on Bax/Bak and can be reproduced by overexpression of Bax. The induction of autophagy coincides with cytochrome c release, and taken together the results sugg...

  11. PUMA- and Bax-induced autophagy contributes to apoptosis

    OpenAIRE

    Yee, K S; Wilkinson, S; James, J; Ryan, K M; Vousden, K H

    2009-01-01

    The p53-inducible BH3-only protein PUMA is a key mediator of p53-dependent apoptosis, and PUMA has been shown to function by activating Bax and mitochondrial outer membrane permeabilization. In this study, we describe an ability of PUMA to induce autophagy that leads to the selective removal of mitochondria. This function of PUMA depends on Bax/Bak and can be reproduced by overexpression of Bax. The induction of autophagy coincides with cytochrome c release, and taken together the results sug...

  12. NOXA-induced alterations in the Bax/Smac axis enhance sensitivity of ovarian cancer cells to cisplatin.

    Directory of Open Access Journals (Sweden)

    Chao Lin

    Full Text Available Ovarian cancer is the most common cause of death from gynecologic malignancy. Deregulation of p53 and/or p73-associated apoptotic pathways contribute to the platinum-based resistance in ovarian cancer. NOXA, a pro-apoptotic BH3-only protein, is identified as a transcription target of p53 and/or p73. In this study, we found that genetic variants of Bcl-2 proteins exist among cisplatin-sensitive and -resistant ovarian cancer cells, and the responses of NOXA and Bax to cisplatin are regulated mainly by p53. We further evaluated the effect of NOXA on cisplatin. NOXA induced apoptosis and sensitized A2780s and SKOV3 cells to cisplatin in vitro and in vivo. The effects were mediated by elevated Bax expression, enhanced caspase activation, release of Cyt C and Smac into the cytosol. Furthermore, gene silencing of Bax or Smac significantly attenuated NOXA and/or cisplatin-induced apoptosis in chemosensitive A2780s cells, whereas overexpression of Bax or addition of Smac-N7 peptide significantly increased NOXA and/or cisplatin-induced apoptosis in chemoresistant SKOV3 cells. To our knowledge, these data suggest a new mechanism by which NOXA chemosensitized ovarian cancer cells to cisplatin by inducing alterations in the Bax/Smac axis. Taken together, our findings show that NOXA is potentially useful as a chemosensitizer in ovarian cancer therapy.

  13. Effects of curcumin on hippocampal Bax and Bcl-2 expression and cognitive function of a rat model of Alzheimer's disease

    Institute of Scientific and Technical Information of China (English)

    Yunliang Wang; Honglei Yin; Jiyu Lou; Bing Han; Xinyue Qin; Fanchao Meng; Shuang Geng; Yajun Liu

    2011-01-01

    We tested the effects of curcumin treatment on a rat model of Alzheimer's disease induced by beta-amlyoid (Aβ1-40) expression. We investigated alterations in the expression of the apoptosis-related genes Bax and Bcl-2 in the hippocampus, as well as changes in the spatial memory and cognitive function of the rats. Reverse transcription-polymerase chain reaction and immunohistochemistry results showed that Bax expression was remarkably decreased and Bcl-2 expression was increased in the rat Alzheimer's disease model after curcumin treatment. Morris water maze results showed that the average time of escape latency was shortened in the curcumin treated model rats. Our study shows that curcumin can significantly improve spatial learning and memory functions in rats with Aβ1-40-induced Alzheimer's disease by modulating Bax and Bcl-2 expression.

  14. Killing of Brain Tumor Cells by Hypoxia-Responsive Element Mediated Expression of BAX

    Directory of Open Access Journals (Sweden)

    Hangjun Ruan

    1999-11-01

    Full Text Available The presence of radioresistant hypoxic cells in human brain tumors limits the overall effectiveness of conventional fractionated radiation therapy. Tumor-specific therapies that target hypoxic cells are clearly needed. We have investigated the expression of suicide genes under hypoxia by a hypoxia-responsive element (HRE, which can be activated through hypoxia-inducible factor-1 (HIF-1. We transfected plasmids containing multiple copies of HIRE into U-87 MG and U-251 MG-NCI human brain tumor cells and tested their ability to induce LacZ gene expression under anoxia. Gene expression under anoxia versus oxia was increased about 12-fold for U-87 MG cells and about fourfold for U-251 MG-NCI cells. At intermediate hypoxic conditions, increased LacZ gene expression in U-87 MG cells was induced by the plasmid that contained three HREs, but not by the plasmid with two HREs. Lastly, when we placed a suicide gene BAX under the control of HREs, cells transfected with the BAX plasmids were preferentially killed through apoptosis under anoxia. Our studies demonstrate that HRE-regulated gene expression is active in brain tumor cells, and that the amount of increased gene expression obtained is dependent on the cell line, the HIRE copy number, and the degree of hypoxia.

  15. Molecular cloning, characterization and expression analysis of (B-cell lymphoma-2 associated X protein) Bax in the orange-spotted grouper (Epinephelus coioides) after the Vibrio alginolyticus challenge.

    Science.gov (United States)

    Luo, Sheng-Wei; Wang, Wei-Na; Sun, Zuo-Ming; Xie, Fu-Xing; Kong, Jing-Rong; Liu, Yuan; Cheng, Chang-Hong

    2016-07-01

    Bax is a pro-apoptotic member of Bcl-2 like superfamily, playing an important role in regulating the apoptosis. In this study, the full-length Bax (EcBax) was obtained, containing a 5'UTR of 64 bp, an ORF of 579 bp and a 3'UTR of 1021 bp. The EcBax gene encoded a polypeptide of 192 amino acids with an estimated molecular mass of 21.55 KDa and a predicted isoelectric point (pI) of 6.75. The deduced amino acid sequence analysis showed that EcBax comprised the conserved residues and the characteristic domains known to the critical function of Bax. qRT-PCR analysis revealed that EcBax mRNA was broadly expressed in all of the examined tissues, while the highest expression level was observed in blood, followed by the expression in liver, gill, spleen, kidney, heart, muscle and intestine. A sharp increase of EcBax expression was observed in the vibrio challenge group by comparing with those in the control. Subcellular localization analysis revealed that EcBax was predominantly localized in the cytoplasm. EcBax exerted a regulatory role in modulating the mitochondrial membrane potential, promoting the cytochrome c release, and then activating the downstream caspase signaling. Moreover, the overexpression of EcBax can decrease the cell viability and antagonize NF-kB, AP-1, Stat3 promoter activity in Hela cells. These results indicate that EcBax containing the conserved domain of pro-apoptotic member of Bcl-2 family may disrupt the mammalian signaling and play a regulative role in the apoptotic process.

  16. A novel plant glutathione S-transferase/peroxidase suppresses Bax lethality in yeast

    DEFF Research Database (Denmark)

    Kampranis, S C; Damianova, R; Atallah, M;

    2000-01-01

    The mammalian inducer of apoptosis Bax is lethal when expressed in yeast and plant cells. To identify potential inhibitors of Bax in plants we transformed yeast cells expressing Bax with a tomato cDNA library and we selected for cells surviving after the induction of Bax. This genetic screen allo...

  17. Structural and Biochemical Studies of TIGAR (TP53-induced Glycolysis and Apoptosis Regulator)

    Energy Technology Data Exchange (ETDEWEB)

    Li, H.; Jogl, G

    2009-01-01

    Activation of the p53 tumor suppressor by cellular stress leads to variable responses ranging from growth inhibition to apoptosis. TIGAR is a novel p53-inducible gene that inhibits glycolysis by reducing cellular levels of fructose-2,6-bisphosphate, an activator of glycolysis and inhibitor of gluconeogenesis. Here we describe structural and biochemical studies of TIGAR from Danio rerio. The overall structure forms a histidine phosphatase fold with a phosphate molecule coordinated to the catalytic histidine residue and a second phosphate molecule in a position not observed in other phosphatases. The recombinant human and zebra fish enzymes hydrolyze fructose-2,6-bisphosphate as well as fructose-1,6-bisphosphate but not fructose 6-phosphate in vitro. The TIGAR active site is open and positively charged, consistent with its enzymatic function as bisphosphatase. The closest related structures are the bacterial broad specificity phosphatase PhoE and the fructose-2,6-bisphosphatase domain of the bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. The structural comparison shows that TIGAR combines an accessible active site as observed in PhoE with a charged substrate-binding pocket as seen in the fructose-2,6-bisphosphatase domain of the bifunctional enzyme.

  18. Combined analysis of cell growth and apoptosis-regulating proteins in HPVs associated anogenital tumors

    International Nuclear Information System (INIS)

    The clinical course of human papillomavirus (HPV) associated with Bowenoid papulosis and condyloma acuminatum of anogenital tumors are still unknown. Here we evaluated molecules that are relevant to cellular proliferation and regulation of apoptosis in HPV associated anogenital tumors. We investigated the levels of telomerase activity, and inhibitor of apoptosis proteins (IAPs) family (c-IAP1, c-IAP2, XIAP) and c-Myc mRNA expression levels in 20 specimens of Bowenoid papulosis and 36 specimens of condyloma acuminatum in anogenital areas. Overall, phosphorylated (p-) AKT, p-ribosomal protein S6 (S6) and p-4E-binding protein 1 (4EBP1) expression levels were examined by immunohistochemistry in anogenital tumors both with and without positive telomerase activity. Positive telomerase activity was detected in 41.7% of Bowenoid papulosis and 27.3% of condyloma acuminatum compared to normal skin (p < 0.001). In contrast, the expression levels of Bowenoid papulosis indicated that c-IAP1, c-IAP2 and XIAP mRNA were significantly upregulated compared to those in both condyloma acuminatum samples (p < 0.001, p < 0.001, p = 0.022, respectively) and normal skin (p < 0.001, p = 0.002, p = 0.034, respectively). Overall, 30% of Bowenoid papulosis with high risk HPV strongly promoted IAPs family and c-Myc but condyloma acuminatum did not significantly activate those genes. Immunohistochemically, p-Akt and p-S6 expressions were associated with positive telomerase activity but not with p-4EBP1 expression. Combined analysis of the IAPs family, c-Myc mRNA expression, telomerase activity levels and p-Akt/p-S6 expressions may provide clinically relevant molecular markers in HPV associated anogenital tumors

  19. Effects of Radix notoginseng extracts drug-containing serum on expressions of bcl-2, Bax and p21WAF1 proteins in MNNG transformed GES-1 cells%三七提取物含药血清对MNNG转化后GES-1细胞凋亡相关基因蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    李军祥; 王志斌; 朱陵群; 牛福玲; 崔巍

    2008-01-01

    Objective: To investigate the effects of Radix notoginseng extracts drug-containing serum on the expressions of apoptosis-regulating proteins including Bax, bcl-2 and p21WAF1 in precancerous gastric cells. Methods: The N-methyI-N'-nitro-N-nitroso-guanidine (MNNG) transformed eternalized human gastric mucosa epithelium GES-I cell line (MC cell) was used in vitro as a model of gastric precancerous lesion. The medicated canine serum was prepared by feeding to the adult Beagle dog with Radix notoginseng extracts and obtaining the serum after 2-hour medication. MC cells were cultured with medicated canine serum (medicated serum group) or non-medicated canine serum (normal control group) for 72 hours. Expressions of Bax, bcl-2 and p21WAF1 proteins were detected by immunocytochemical assay and the average optical density of the cells was determined by an image analysis system. Results. Compared with those of the normal control group, Bax and p21WAF1 expressions in medicated serum group were significantly enhanced (P<0.01), while the expression of bcl-2 was significantly reduced (P 001). Conclusion. Radix notoginseng extracts may inhibit the proliferation and promote the apoptosis of precancerous gastric cells through altering expressions of the bcl-2, Bax and p21WAF1 genes.%目的:研究三七提取物犬药物血清作用于胃癌前细胞后,其凋亡相关基因蛋白表达的变化.方法:采用被N-甲基-N-硝基-N-亚硝基胍(N-methyl-N-nitroso-guanidine,MNNG)转化后的永生化人胃黏膜上皮细胞系GES-1细胞(简称MC细胞)作为胃癌前病变细胞的体外研究模型,用三七提取物一次性灌胃彼格犬,取给药后2 h的血清作为实验药物血清.以免疫组织化学法检测药物血清对MC细胞作用72 h后bcl-2、Bax和p21WAF13种凋亡相关基因蛋白表达情况,并与正常培养的MC细胞相比较.结果:药物血清作用后的MC细胞中Bax和p21WAF1的表达较正常培养的MC细胞升高(P<0.01);Bc1-2表达较

  20. New molecular analysis of differential gene expressions to evaluate new exposure markers and radioresistance

    International Nuclear Information System (INIS)

    Molecular techniques, such as macro array and representation difference analysis (RDA) (1), allow to detect subtle variations into complex biological processes induced by exposure to ionising radiation. One of the most reliable method to investigate radioresistance in vitro is to select a clone with acquired or intrinsic resistant phenotype by delivering repeated fractions of low-dose X-irradiation to a parent cell line. The resulting isolated resistant clone is then suitable for molecular techniques to analyse differential genes which expressions are important in characterising response and resistance to radiation. The cDNA expression arrays allow to perform the analysis of hundreds of known genes while RDA permits the comparison of genomic cDNA also from higher eukaryotes. The aim of the present work was to verify the suitable of these new molecular approaches to recognize the expression of genes hypothetically useful as radioprotection markers. To this end, a relatively high dose of X-rays was used (2 Gy), differentially expressed genes were isolated, and new experiments based on high sensitive and reproducible RT-PCR are foreseen for lower doses. Human neuroblastoma cell lines IMR32 and its resistant clone (Clone F), previously isolated by repeated 2 Gy X-irradiation( 2), were irradiated with a single 2 Gy X-rays. Six hours later, cells were monitored for surviving fraction, index of apoptosis and RNAs were extracted, purified and analysed either by human macro array with 205 cDNA of apoptosis genes related spiked on and either by RDA methodology. Human apoptosis macro array confirmed higher expression of genes related both to apoptosis regulator (Bax) and apoptosis effectors (caspase-2) in IMR32 cell line. RDA showed several differentially expressed genes in the resistant clone. Among these genes, two unknown forms of a protein with a putative enzymatic activity are cloned and transfected in the sensitive cell line to understand their role in radioresistance

  1. EGR-1 forms a complex with YAP-1 and upregulates Bax expression in irradiated prostate carcinoma cells.

    Science.gov (United States)

    Zagurovskaya, M; Shareef, M M; Das, A; Reeves, A; Gupta, S; Sudol, M; Bedford, M T; Prichard, J; Mohiuddin, M; Ahmed, M M

    2009-02-26

    In this study, we investigated the functional role of early growth response-1 (Egr1 gene) in the regulation of radiation-induced clonogenic inhibition and apoptosis in p53 wild-type and mutant prostate cancer cells 22Rv1 and DU145, respectively. 22Rv1 cells were more sensitive to irradiation compared with DU145 cells, and the sensitivity was enhanced by overexpression of EGR-1 in both cells. Dominant-negative EGR-1 mutant (dnEGR-1) or repressor of EGR-1, NGFIA binding protein 1 (NAB1), increased radioresistance of these cells. Significant activation of caspases 3 and 9 and Bcl2-associated X (Bax) with increased poly(ADP-ribose) polymerase (PARP) cleavage and cytochrome c release was observed in radiation-exposed EGR-1 overexpressing cells. Gel shift analysis and chloramphenicol acetyl transferase (CAT) reporter assays indicate that EGR-1 transactivates the promoter of the Bax gene. Interaction of EGR-1 and Yes kinase-associated protein 1 (YAP-1) through the WW domain of YAP-1 enhances the transcriptional activity of EGR-1 on the Bax promoter as shown by chromatin immunoprecipitation and reporter assays. Irradiation of PC3 cell xenografts that were treated with adenoviral EGR-1 showed significant regression in tumor volume. These findings establish the radiation-induced pro-apoptotic action of EGR-1, in a p53-independent manner, by directly transactivating Bax, and prove that alters the B-cell CLL/lymphoma 2 (Bcl-2)/Bax ratio as one of the mechanisms resulting in significant activation of caspases, leading to cell death through the novel interaction of EGR-1 with YAP-1. PMID:19137013

  2. EXPRESSION OF BAX AND BCL-2 IN MOUSE OFFSPRING BRAIN AFTER MATERNAL ORAL ADMINIS TRATION OF MONOSODIUM GLUTAMATE

    Institute of Scientific and Technical Information of China (English)

    徐磊; 赵晏; 展淑琴; 王会生; 史文春

    2002-01-01

    Objective To analyze the excitotoxicity of monoso dium glutamate (MSG) in the offspring cerebral cortex and hippocampal subregions after maternal oral administration of MSG. Methods Kunming mi ce were given per os MSG ( 4.0 g/kg ) at 17~21 days of pregnancy and their offs pring behaviors were studied at 10, 20 , 30 days postnatally. By using immunohis tochemical means, the involvement of Bcl-2 and Bax in the glutamate-induced c ell death in cortical and hippocampal neur ons were examined. Cell damage was assessed by direct cell counting. Res ults Administration of monosodium glutamate during the fetal period in mice resulted in a moderate increase in the expression of Bax in principal neuro ns in CA1, CA2, CA3, CA4 and in the cerebral cortex at postpartum 10, 20, 30 day s in the offspring mice, whereas Bcl-2 protein expressions were reduced signif icantly in the same regions as compared with those of controls. Conclusi on These findings suggest that glutamate toxicity results in cellular d eath via an apoptotic mechanism in which the Bcl-2/Bax-alpha molecular comple x may be involved. The glutamate-induced apoptosis appears to be related to the modulation of Bcl-2 family gene products such as Bcl-2 and Bax.

  3. EXPRESSION OF BAX AND BCL-2 IN MOUSE OFFSPRING BRAIN AFIER MATERNAL ORAL ADMINISTRATION OF MONOSODIUM GLUTAMATE

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objective:To analyze the excitotoxicity of monosodium glutamate(MSG)in the offspring crebral cortex and hippocampal subresions after maternal oral administration of MSG.Methods:Kunming mice were given per os MSG(4.0g/kg)at 17-21 days of pregnancy and their offspring behaviors were studied at 10,20,30days postnatally.By using inmunohistochemical means,the involvment of Bcl-2 and bax in the glutamate-induced cell death in cortical and hippocampal neurons were examined.Cell damage was assessed by direct cell counting.Results:administration of monosodium glutamate during the fetal period in mice resulted in a moderate increase in the expression of Bax in principal neurons in CA1,CA2,CA3,CA4 and in the cerebral cortex at postpartum 10,20,30 days in the offspring mice,whereas Bcl-2 protein expressions were reduced significantly in the same regions as compared with those of controls.Conclusion:These findings suggest that glutamate toxicity results in cellular death via an apoptotic mechanism in which the Bcl-2/Bax-alpha molecular complex may be involved.The glutamate-induced apoptosis appears to be related to the modulation of Bcl-2 family gene products such as Bcl-2 and Bax.

  4. Upregulation of Bax and Bcl-2 following prenatal cocaine exposure induces apoptosis in fetal rat brain

    Directory of Open Access Journals (Sweden)

    DaLiao Xiao, Lubo Zhang

    2008-01-01

    Full Text Available Cocaine abuse during pregnancy has been associated with numerous adverse perinatal outcomes. Aims: The present study was to determine whether prenatal cocaine exposure induced apoptosis and the possible role of Bcl-2 family genes in the programming cell death in fetal rat brain. Main methods: Pregnant rats were treated with cocaine subcutaneously (30 & 60 mg/kg/day from day 15 to 21 of gestation. Then the fetal and maternal brains were isolated. Key findings: Cocaine produced a dose-dependent decrease in fetal brain weight and brain/body weight ratio (P<0.05. Apoptotic nuclei in fetal brain were increased from 2.6 ± 0.1 (control to 8.1± 0.6 (low dose and 10.4 ± 0.2% (high dose (P<0.05. In accordance, cocaine dose dependently increased activities of caspase-3, caspase-8, and caspase-9 (% of control in the fetal brain by 177%, 155%, 174%, respectively, at 30 mg/kg/day, and by 191%, 176%, 274%, respectively, at 60 mg/kg/day. In contrast, cocaine showed no effect on caspase activities in the maternal brain. Cocaine produced a dose-dependent increase in both Bcl-2 and Bax protein expression in the fetal brain, and increased the ratio of Bax/Bcl-2 at dose of 30 mg/kg/day (P<0.05. Significance: Our study has demonstrated that prenatal cocaine exposure induces apoptosis in the fetal brain, and suggested that up-regulating Bax/Bcl-2 gene expression may be involved in cocaine-induced apoptosis. The increased apoptosis of neuronal cells in the fetal brain is likely to play a key role in cocaine-induced neuronal defects during fetal development.

  5. Enhancing survival of mouse oocytes following chemotherapy or aging by targeting Bax and Rad51.

    Directory of Open Access Journals (Sweden)

    Loro L Kujjo

    Full Text Available BACKGROUND: Therapeutic approaches to preserve fertility in females undergoing cancer treatments are currently ineffective. This is partly due to limited knowledge of the molecular mechanisms that injured germ cells elicit to repair damage and survive or to abort repair and activate biochemical pathways leading to death. So far, we know that following spontaneously occurring or drug-induced DNA damage, the efficiency of DNA repair is a critical determinant of the cell's fate. The protein encoded by the Rad51 gene is one of several components recruited for homologous recombination-dependent DNA double-strand break repair in both somatic cells and germ cells. Recently, we showed that microinjection of recombinant Rad51 into AKR/J mouse oocytes decreased the extent of spontaneous DNA double-strand breaks, suppressed apoptosis, and restored the developmental competence in AKR/J embryos. Herein we characterized the nature of chemotherapy-induced lesions in oocytes, and the associated individual components of the DNA damage sensor and repair apparatus. For comparison, we also assessed parallel spontaneous changes in aging oocytes. METHODS: Data collected were derived from: analysis of apoptosis; immunodepletion; oocyte microinjections; immunocytochemistry; immunofluorescence; and CHIP-like assays. RESULTS: Our data show that: (i DNA damage in oocytes can be induced by both chemotherapy and spontaneously by the aging process; (ii oocytes possess the machinery and capability for repairing such DNA damage; (iii Rad51 is a critical player in the repair of both chemotherapy-induced and spontaneously-sustained DNA damage; and (iv in response to damage, oocytes exhibit an inverse functional relationship between presence of Bax and activity of Rad51. CONCLUSION/SIGNIFICANCE: Our results establish Rad51 and/or Bax as potential candidates that can be targeted for development of individualized chemotherapeutic interventions that are effective, but minimal in

  6. NFAT2 mediates high glucose-induced glomerular podocyte apoptosis through increased Bax expression

    Energy Technology Data Exchange (ETDEWEB)

    Li, Ruizhao, E-mail: liruizhao1979@126.com [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China); Zhang, Li, E-mail: Zhanglichangde@163.com [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China); Southern Medical University, Guangzhou, Guangdong (China); Shi, Wei, E-mail: shiwei.gd@139.com [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China); Zhang, Bin, E-mail: zhangbinyes@yahoo.com.cn [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China); Liang, Xinling, E-mail: xinlingliang@yahoo.com [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China); Liu, Shuangxin, E-mail: mplsxi@yahoo.com.cn [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China); Wang, Wenjian, E-mail: wwjph@yahoo.com [Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan No. 2 Road, Guangzhou, 510080 (China)

    2013-04-15

    Background: Hyperglycemia promotes podocyte apoptosis and plays a key role in the pathogenesis of diabetic nephropathy. However, the mechanisms that mediate hyperglycemia-induced podocyte apoptosis is still far from being fully understood. Recent studies reported that high glucose activate nuclear factor of activated T cells (NFAT) in vascular smooth muscle or pancreatic β-cells. Here, we sought to determine if hyperglycemia activates NFAT2 in cultured podocyte and whether this leads to podocyte apoptosis. Meanwhile, we also further explore the mechanisms of NFAT2 activation and NFAT2 mediates high glucose-induced podocyte apoptosis. Methods: Immortalized mouse podocytes were cultured in media containing normal glucose (NG), or high glucose (HG) or HG plus cyclosporine A (a pharmacological inhibitor of calcinerin) or 11R-VIVIT (a special inhibitor of NFAT2). The activation of NFAT2 in podocytes was detected by western blotting and immunofluorescence assay. The role of NFAT2 in hyperglycemia-induced podocyte apoptosis was further evaluated by observing the inhibition of NFAT2 activation by 11R-VIVIT using flow cytometer. Intracellular Ca{sup 2+} was monitored in HG-treated podcocytes using Fluo-3/AM. The mRNA and protein expression of apoptosis gene Bax were measured by real time-qPCR and western blotting. Results: HG stimulation activated NFAT2 in a time- and dose-dependent manner in cultured podocytes. Pretreatment with cyclosporine A (500 nM) or 11R-VIVIT (100 nM) completely blocked NFAT2 nuclear accumulation. Meanwhile, the apoptosis effects induced by HG were also abrogated by concomitant treatment with 11R-VIVIT in cultured podocytes. We further found that HG also increased [Ca{sup 2+}]i, leading to activation of calcineurin, and subsequent increased nuclear accumulation of NFAT2 and Bax expression in cultured podocytes. Conclusion: Our results identify a new finding that HG-induced podocyte apoptosis is mediated by calcineurin/NFAT2/Bax signaling pathway

  7. Melatonin restores normal Bax and Bcl-2 protein expression in the subgranular zone of the dentate gyrus in pinealectomized rats

    Institute of Scientific and Technical Information of China (English)

    Shengchang Zhang; Shuang Zhao; Lu Bai; Mingming Guan; Jielin Mo; Ling Lan

    2011-01-01

    In this study, we sought to elucidate the effects of melatonin on learning and memory as well as apoptosis and expression of the Bax or Bcl-2 proteins in the subgranular zone of the dentate gyrus in pinealectomized rats. Using the Morris water maze and the olfactory memory tests, we found that the average escape latency in pinealectomized rats was clearly increased compared with sham-operated rats. Moreover, the average escape latency in the melatonin-treated and pinealectomized rats was longer than that in the sham-operated rats and shorter than that in the pinealectomized and untreated rats. Immunohistochemistry and terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) showed that there were fewer Bax immunoreactive cells and TUNEL-positive (apoptotic) cells but more Bcl-2 immunoreactive cells in the melatonin-treated rats compared with the pinealectomized rats. The sham-operated rats showed numbers of these cells similar to the melatonin-treated rats. These experimental findings demonstrate that melatonin treatment may reduce abnormal apoptosis by promoting gene expression of Bax and suppressing gene expression of Bcl-2 in the subgranular zone of the dentate gyrus in pinealectomized rats. These effects appear to result in the inhibition of cellular apoptosis and the improvement of spatial learning and memory in pinealectomized rats.

  8. Virosecurinine induces apoptosis by affecting Bcl-2 and Bax expression in human colon cancer SW480 cells.

    Science.gov (United States)

    Chen, Chuan-Rong; Xia, Yong-Hui; Yao, Shu-Yan; Zhang, Qing; Wang, Ying; Ji, Zhao-Ning

    2012-04-01

    Virosecurinine, the major alkaloid isolated from Securinega suffruticosa Pall Rehd was found to exhibit growth inhibition and cytotoxicity against huaman colon cancer SW480 cells via the microculture tetrazolium (MTT) assay. Due to its greater cytotoxic potency and selectivity towards SW480 cells, flow cytometry was used to analyze the cell cycle distribution of control and treated SW480 cells whereas Annexin V-FITC/PI flow cytometry analysis was carried out to confirm apoptosis induced by virosecurinine in SW480 cells. Apoptotic regulatory genes were determined by RT-PCR analysis. Virosecurinine was found to induce G1/S cell cycle arrest which led to predominantly apoptotic mode of cell death. Mechanistically, virosecurinine was found to up-regulated the Bax gene expression and down-regulated the Bcl-2 expression in SW480, The ratio of Bcl-2 to Bax was significantly decreased. Hence, we suggest that virosecurinine induced apoptosis in SW480 cells by affecting the expression of bcl-2 and bax. PMID:22570942

  9. Spatial and temporal changes in Bax subcellular localization during NPe6-PDT-induced apoptosis

    Science.gov (United States)

    Liu, Lei; Xing, Da; Chen, Wei R.; Wan, Qingling; Zhou, Feifan

    2008-02-01

    Photodynamic therapy (PDT) employing photosensiter N-aspartyl chlorin e6 (NPe6) can induce lysosome disruption and initiate the intrinsic apoptotic pathway. Bax, a member of the Bcl-2 family of proteins, is an essential regulator of apoptosis. Bax is normally found in the cytosol of healthy cells, and translocates to mitochondria in response to many apoptotic stimuli. In this study, using real-time single-cell analysis, we have investigated the kinetics of Bax distribution during NPe6-induced apoptosis in ASTC-a-1 cells. In order to monitor Bax subcellular distribution, cells were stained with GFP-Bax and Mito Tracker Red. The results show that Bax redistribution occurred at about 170 min after treated with NPe6-PDT, and then sequestered into clusters associated with the mitochondira within 30 min. Our data clearly showed the spatial and temporal changes in Bax distribution in living cells during NPe6-induced apoptosis.

  10. Clinicopathological significance of Bcl-2 and Bax protein expression in human pancreatic cancer

    Institute of Scientific and Technical Information of China (English)

    Ming Dong; Jian-Ping Zhou; Hao Zhang; Ke-Jian Guo; Yu-Lin Tian; Yu-Ting Dong

    2005-01-01

    AIM: To assess the clinicopathological significance of the expression of the apoptosis-inhibitory Bcl-2 protein (pBcl-2) and the apoptosis-promoting Bax protein (pBax) in human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: Fifty-nine surgical specimens of IDCs of the pancreas were stained immunohistochemically to detectpBcl-2 and pBax expressions whose correlation to tumor classification, staging, and prognosis was analyzed by univariate and multivariate analyses. RESULTS: The expression of pBcl-2 and pBax was detected in 21 of 59 (35.6%) and in 29 of 59 (49.2%) patients with IDCs of the pancreas, respectively. Neither pBcl-2 nor pBax alone was correlated to TNM staging and differentiation degree of IDCs of the pancreas according to univariate analysis. By Mantel-Cox test, the median survival time after surgery for pBcl-2(+) and pBcl-2(-) groups were 14.3 and 7.3 mo, respectively (χ2= 9.357, P = 0.002) and that for pBax(+) and pBax(-) groups were 12.9 and 10.2 mo, respectively (χ2= 0.285, P>0.05).Contingency coefficient between pBd-2 and pBax expression was 0.298, indicating that there was correlation between them (χ2= 5.74, P<0.05). The median survival time after surgery for pBd-2(+)pBax(+) and pBcl-2(+)pBax(-) groups were 14.3 and 14.1 mo, respectively, and that for pBcl-2 (-)pBax(+) and pBcl-2(-)pBax(-) groups were 5.9 and 9.9 mo, respectively. There was a significant difference between pBcl-2(+)pBax(+) and pBcl-2(-)pBax(+) (χ2 = 5.06,P<0.05), such was the case for pBcl-2(+)pBax(+) andpBcl-2(-)pBax(-) (χ2= 7.18, P<0.01). Cox proportional hazards model for multivariate analysis was applied, indicating that pBcl-2, TNM staging, age and pBax were high risk factors of post-surgical survival time. CONCLUSION: Both pBcl-2 and pBax have high expression in IDCs of the pancreas, indicating that co-expression of pBcl-2 and pBax is a good indicator of favorable prognosis in IDCs of the pancreas.

  11. E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation

    International Nuclear Information System (INIS)

    PUMA is a pro-apoptotic Bcl-2 family member that has been shown to be involved in apoptosis in many cell types. We sought to ascertain whether induction of PUMA plays a crucial role in E2F-1-induced apoptosis in melanoma cells. PUMA gene and protein expression levels were detected by real-time PCR and Western blot in SK-MEL-2 and HCT116 cell lines after Ad-E2F-1 infection. Activation of the PUMA promoter by E2F-1 overexpression was detected by dual luciferase reporter assay. E2F-1-induced Bax translocation was shown by immunocytochemistry. The induction of caspase-9 activity was measured by caspase-9 colorimetric assay kit. Up-regulation of the PUMA gene and protein by E2F-1 overexpression was detected by real-time PCR and Western blot analysis in the SK-MEL-2 melanoma cell line. In support of this finding, we found six putative E2F-1 binding sites within the PUMA promoter. Subsequent dual luciferase reporter assay showed that E2F-1 expression could increase the PUMA gene promoter activity 9.3 fold in SK-MEL-2 cells. The role of PUMA in E2F-1-induced apoptosis was further investigated in a PUMA knockout cell line. Cell viability assay showed that the HCT116 PUMA-/- cell line was more resistant to Ad-E2F-1-mediated cell death than the HCT116 PUMA+/+ cell line. Moreover, a 2.2-fold induction of the PUMA promoter was also noted in the HCT116 PUMA+/+ colon cancer cell line after Ad-E2F-1 infection. Overexpression of a truncated E2F-1 protein that lacks the transactivation domain failed to up-regulate PUMA promoter, suggesting that PUMA may be a transcriptional target of E2F-1. E2F-1-induced cancer cell apoptosis was accompanied by Bax translocation from the cytosol to mitochondria and the induction of caspase-9 activity, suggesting that E2F-1-induced apoptosis is mediated by PUMA through the cytochrome C/Apaf-1-dependent pathway. Our studies strongly demonstrated that E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation. The signaling

  12. Expression of Bax in yeast affects not only the mitochondria but also vacuolar integrity and intracellular protein traffic

    DEFF Research Database (Denmark)

    Dimitrova, Irina; Toby, Garabet G; Tili, Esmerina;

    2004-01-01

    Bax-induced lethality in yeast is accompanied by morphological changes in mitochondria, giving rise to a reduced number of swollen tubules. Although these changes are completely abolished upon coexpression of the Bax inhibitor, Bcl-2, coexpression of Bax with Bax inhibiting-glutathione S-transfer...

  13. 8周中等强度低负荷量训练对老龄雌性大鼠骨骼肌Bax和Bcl-2蛋白及SIRT1/SIRT3信号轴基因表达的影响%Effects of 8-week medium intensity low load training on proteins Bax and Bcl-2 and the gene expression of signal axis SIRT1/SIRT3 of skeletal muscle of aged female rats

    Institute of Scientific and Technical Information of China (English)

    李方晖; 肖琳; 覃飞; 刘承宜

    2014-01-01

    In order to observe the effects of 8-week medium intensity low load training on the levels of proteins Bax and Bcl-2 and the gene messenger RNA (mRNA) expression of axis sirtuin 1 (SIRT1)/sirtuin 3 (SIRT3) of gastrocne-mius of aged rats, the authors divided 16 18-month old female SD rats randomly into a control group and an exercise group, each of which contained 8 rats, let the rats in the exercise group do an aerobic exercise on a treadmill for con-secutive 8 weeks, at a speed of 15 km/h (with 60%~75%VO2max), 15 minutes a day, 5 days a week, let the rats in the control group live freely, in 24 hours after rat exercising at the end of week 8, killed the rats, measured gastrocnemius index, measured the levels of proteins Bax and Bcl-2 of gastrocnemius by means of Western blot analysis, measured the mRNA levels of SIRT3, SIRT1, manganese superoxide dismutase (MnSOD), Caspase 3, peroxisome prolifera-tor-activated receptor-γcoactivator-1 (PGC-1α), mitochondrial transcription factor A (TFAM) and nuclear respiratory factor 1 (NRF1) by means of RT-PCR, and revealed the following findings:as for the rats in the exercise group, their gastrocnemius mass and gastrocnemius index increased significantly (P<0.05 and P<0.01 respectively), their protein Bax level decreased significantly (P<0.05), their protein Bcl-2 level and Bcl-2/Bax ratio increased significantly (P<0.05);their mRNA levels of SIRT3, SIRT1, PGC-1α, NRF1, TFAM and MnSOD increased significantly (P<0.05), their mRNA level of Caspase-3 decreased significantly (P<0.05). The said findings indicated the followings:medium intensity low load training could delay the changing of muscle cell apoptosis signal of aged rats; the homeostatic mechanism mediated by axis SIRT1/SIRT3 played an important role in medium intensity low load training increasing the mitochondria refreshing rate and antioxidase level of skeletal muscle of aged rats.%观察8周中等强度低负荷量训练对老龄雌性大鼠腓肠肌Bax和Bcl-2

  14. Bax function in the absence of mitochondria in the primitive protozoan Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Adrian B Hehl

    Full Text Available Bax-induced permeabilization of the mitochondrial outer membrane and release of cytochrome c are key events in apoptosis. Although Bax can compromise mitochondria in primitive unicellular organisms that lack a classical apoptotic machinery, it is still unclear if Bax alone is sufficient for this, or whether additional mitochondrial components are required. The protozoan parasite Giardia lamblia is one of the earliest branching eukaryotes and harbors highly degenerated mitochondrial remnant organelles (mitosomes that lack a genome. Here we tested whether human Bax expressed in Giardia can be used to ablate mitosomes. We demonstrate that these organelles are neither targeted, nor compromised, by Bax. However, specialized compartments of the regulated secretory pathway are completely ablated by Bax. As a consequence, maturing cyst wall proteins that are sorted into these organelles are released into the cytoplasm, causing a developmental arrest and cell death. Interestingly, this ectopic cargo release is dependent on the carboxy-terminal 22 amino acids of Bax, and can be prevented by the Bax-inhibiting peptide Ku70. A C-terminally truncated Bax variant still localizes to secretory organelles, but is unable to permeabilize these membranes, uncoupling membrane targeting and cargo release. Even though mitosomes are too diverged to be recognized by Bax, off-target membrane permeabilization appears to be conserved and leads to cell death completely independently of mitochondria.

  15. Bax function in the absence of mitochondria in the primitive protozoan Giardia lamblia.

    Science.gov (United States)

    Hehl, Adrian B; Regos, Attila; Schraner, Elisabeth; Schneider, André

    2007-01-01

    Bax-induced permeabilization of the mitochondrial outer membrane and release of cytochrome c are key events in apoptosis. Although Bax can compromise mitochondria in primitive unicellular organisms that lack a classical apoptotic machinery, it is still unclear if Bax alone is sufficient for this, or whether additional mitochondrial components are required. The protozoan parasite Giardia lamblia is one of the earliest branching eukaryotes and harbors highly degenerated mitochondrial remnant organelles (mitosomes) that lack a genome. Here we tested whether human Bax expressed in Giardia can be used to ablate mitosomes. We demonstrate that these organelles are neither targeted, nor compromised, by Bax. However, specialized compartments of the regulated secretory pathway are completely ablated by Bax. As a consequence, maturing cyst wall proteins that are sorted into these organelles are released into the cytoplasm, causing a developmental arrest and cell death. Interestingly, this ectopic cargo release is dependent on the carboxy-terminal 22 amino acids of Bax, and can be prevented by the Bax-inhibiting peptide Ku70. A C-terminally truncated Bax variant still localizes to secretory organelles, but is unable to permeabilize these membranes, uncoupling membrane targeting and cargo release. Even though mitosomes are too diverged to be recognized by Bax, off-target membrane permeabilization appears to be conserved and leads to cell death completely independently of mitochondria. PMID:17534438

  16. Expression of Bcl-2 and Bax in extrahepatic biliary tract carcinoma and dysplasia

    Institute of Scientific and Technical Information of China (English)

    Sheng-Mian Li; Shu-Kun Yao; Nobuyoshi Yamamura; Toshitsugu Nakamura

    2003-01-01

    AIM: To compare the difference of expression of Bcl-2 and Bax in extrahepatic biliary tract carcinoma and dysplasia, and to analyze the role of Bcl-2 and Bax proteins in the progression from dysplasia to carcinoma and to evaluate the correlation of Bcl-2/Bax protein expression with the biological behaviors.METHODS: Expressions of Bcl-2 and Bax were examined immunohistochemically in 27 cases of extrahepatic biliary tract carcinomas (bile duct carcinoma: n=21, carcinoma of ampulla of Vater: n=6), and 10 cases of atypical dysplasia.Five cases of normal biliary epithelial tissues were used as controls. A semiquantitative scoring system was used to assess the Bcl-2 and Bax reactivity.RESULTS: The expression of Bd-2 was observed in 10 out of 27 (37.0 %) invasive carcinomas, 1 out of 10 clysplasias, none out of 5 normal epithelial tissues. Bax expression rate was 74.1% (20/27) in invasive carcinoma, 30 % (3/10) in dysplasia,and 40 % (2/5) in normal biliary epithelium. Bcl-2 and Bax activities were more intense in carcinoma than in dysplasia,with no significant difference in Bcl-2 expression (P=0.1:10),and significant difference in Bax expression (P=0.038). Level of Bax expression was higher in invasive carcinoma than in dysplasia and normal tissue (P=0.012). Bcl-2 expression was correlated to Bax expression (P=0.0059). However, Bcl-2/Bax expression had no correlation with histological subtype,grade of differentiation, or level of invasion.CONCLUSION: Increased Bcl-2/Bax expression from dysplasia to invasive tumors supports the view that this is the usual route for the development of extrahepatic biliary tract carcinoma. Bcl-2/Bax may be involved, at least in part,in the apoptotic activity in extrahepatic biliary carcinoma.

  17. Evidence that inhibition of BAX activation by BCL-2 involves its tight and preferential interaction with the BH3 domain of BAX

    Institute of Scientific and Technical Information of China (English)

    Bonsu Ku; Chengyu Liang; Jae U Jung; Byung-Ha Oh

    2011-01-01

    Interactions between the BCL-2 family proteins determine the cell's fate to live or die. How they interact with each other to regulate apoptosis remains as an unsettled central issue. So far, the antiapoptotic Bc1-2 proteins are thought to interact with BAX weakly, but the physiological significance of this interaction has been vague.Herein, we show that recombinant BCL-2 and BCL-w interact potently with a BCL-2 homology (BH) 3 domain-containing peptide derived from BAX, exhibiting the dissociation constants of 15 and 23 nM, respectively. To clarify the basis for this strong interaction, we determined the three-dimensional structure of a complex of BCL-2 with a BAX peptide spanning its BH3 domain. It revealed that their interactions extended beyond the canonical BH3 domain and involved three nonconserved charged residues of BAX. A novel BAX variant, containing the alanine substitution of these three residues, had greatly impaired affinity for BCL-2 and BCL-w, hut was otherwise indistinguishable from wild-type BAX. Critically, the apoptotic activity of the BAX variant could not be restrained by BCL-2 and BCL-w, pointing that the observed tight interactions are critical for regulating BAX activation. We also comprehensively quantified the binding affinities between the three BCL-2 subfamily proteins. Collectively, the data show that due to the high affinity of BAX for BCL-2, BCL-w and A1, and of BAK for BCL-XL, MCL-1 and A1, only a subset of BH3-only proteins, commonly including BIM, BID and PUMA, could he expected to free BAX or BAK from the antiapoptotic BCL-2 proteins to elicit apoptosis.

  18. Bax and Bak expression in cervical smears of women with low-and high-risk HPV types: A study of 120 cases

    Directory of Open Access Journals (Sweden)

    Eirini Klapsinou

    2015-01-01

    Full Text Available Background: Human papillomavirus (HPV is known to be involved in the carcinogenesis of squamous cells in uterine cervix cancer, mostly by binding and inactivating the p53 and pRb tumor suppressor genes. Lately, evidence has emerged suggesting that HPV oncoproteins may interact with proteins involved in cellular apoptosis as well. Aim: This study aimed to investigate the expression of proapoptotic proteins Bax and Bak in women with low-risk and high-risk HPV types as opposed to HPV-negative women, and in women with normal pap smear compared to women with abnormal Papanicolau test (Pap smear. Materials and Methods: A total of 120 liquid-based cervical samples were subtyped for HPV types with microarray hybridization and then stained and evaluated immunocytochemically for Bax and Bak expression. Statistical analysis was performed on the Bax and Bak scores (percentage of positive cells × staining intensity, the overall percentage of positive cells, and the most prevalent staining intensity group found in each sample. Results: A weak association between negative Bax staining and cytologically normal Pap smears was discovered, whereas cytologically abnormal samples tended to stain weakly or moderately positive. No other statistically significant difference was found in the other analyzed parameters. Conclusion: Cytologically normal pap smears seem to have a slight tendency to stain negative for Bax as opposed to cytologically abnormal pap smears. Although the association is weak, it is an indication that there might be a connection between the expression of Bax and the development of cervical intraepithelial dysplasia, which warrants further investigation in larger-scale studies.

  19. The oxidized phospholipid PazePC promotes permeabilization of mitochondrial membranes by Bax.

    Science.gov (United States)

    Lidman, Martin; Pokorná, Šárka; Dingeldein, Artur P G; Sparrman, Tobias; Wallgren, Marcus; Šachl, Radek; Hof, Martin; Gröbner, Gerhard

    2016-06-01

    Mitochondria play a crucial role in programmed cell death via the intrinsic apoptotic pathway, which is tightly regulated by the B-cell CLL/lymphoma-2 (Bcl-2) protein family. Intracellular oxidative stress causes the translocation of Bax, a pro-apoptotic family member, to the mitochondrial outer membrane (MOM) where it induces membrane permeabilization. Oxidized phospholipids (OxPls) generated in the MOM during oxidative stress directly affect the onset and progression of mitochondria-mediated apoptosis. Here we use MOM-mimicking lipid vesicles doped with varying concentrations of 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine (PazePC), an OxPl species known to significantly enhance Bax-membrane association, to investigate three key aspects of Bax's action at the MOM: 1) induction of Bax pores in membranes without additional mediator proteins, 2) existence of a threshold OxPl concentration required for Bax-membrane action and 3) mechanism by which PazePC disturbs membrane organization to facilitate Bax penetration. Fluorescence leakage studies revealed that Bax-induced leakage, especially its rate, increased with the vesicles' PazePC content without any detectable threshold neither for OxPl nor Bax. Moreover, the leakage rate correlated with the Bax to lipid ratio and the PazePC content. Solid state NMR studies and calorimetric experiments on the lipid vesicles confirmed that OxPl incorporation disrupted the membrane's organization, enabling Bax to penetrate into the membrane. In addition, 15N cross polarization (CP) and insensitive nuclei enhanced by polarization transfer (INEPT) MAS NMR experiments using uniformly (15)N-labeled Bax revealed dynamically restricted helical segments of Bax embedded in the membrane, while highly flexible protein segments were located outside or at the membrane surface. PMID:26947183

  20. 辛伐他汀诱导人胃癌SGC7901细胞凋亡及其对Bax和Bcl-2表达的影响%Effect of Simvastatin on apoptosis and expressions of Bax and Bcl-2 gene in human gastric cancer cell line SGC7901

    Institute of Scientific and Technical Information of China (English)

    朱梦霞; 王芳; 谢娟; 熊文昊; 杨璐; 黄靓

    2015-01-01

    目的 研究辛伐他汀对人胃低分化腺癌细胞株SGC7901凋亡的影响及其可能的分子机制.方法 体外培养人胃癌SGC7901细胞至对数生长期, 再分别用不同浓度的辛伐他汀处理SGC7901细胞,48h后采用流式细胞术检测细胞凋亡;RT-PCR法和Western Blot法观察Bax和Bcl-2的表达.结果 辛伐他汀能诱导SGC7901细胞凋亡,且呈浓度依赖性.流式细胞术检测显示10、20、40μmol/L辛伐他汀组细胞凋亡率分别为(20.37±3.60)%、(35.17±3.91)%、(58.39±4.06)%,与对照组(4.78±1.51)%相比,凋亡率显著增强(P<0.05).不同浓度的辛伐他汀作用后能显著增强SGC7901细胞Bax mRNA和蛋白的表达,降低Bcl-2 mRNA和蛋白的表达(P<0.05). 结论 辛伐他汀呈浓度依赖性地诱导胃癌SGC7901细胞凋亡,其机制可能与上调Bax、下调Bcl-2表达有关.%Objective To investigate the effects of Simvastatin on apoptosis in human gastric lower-differentiation ade-nocarcinoma cell line SGC7901 and to explore its potential molecular mechanisms. Methods Gastric SGC7901 cells were cultured in vitro, cells of exponential phase of growth were used to experiment. Then using different concentrations of Simvas-tatin role in SGC7901, 48 hours later, flow cytometry method was used to detect the cell apoptosis, the expressions of Bax and Bcl-2 were tested by RT-PCR and Western-Blot assay. Results Simvastatin could promote the apoptosis of gastric cell line SGC7901 in a dose-dependent manner. Flow cytometry method showed that,Simvastatin (10μmol/L, 20μmol/L, 40μmol/L) role in SGC7901 for 48 hours, the apoptosis rates of the three groups were (20.37±3.60)%,(35.17±3.91)%,(58.39±4.06)%, the apoptosis rate of the control group was (4.78±1.51)%, the experimental groups were significantly higher than the control group (P<0.05). Different concentrations of Simvastatin could promote the expression of mRNA and protein of Bax, reduce the expres-sion of mRNA and protein of Bcl-2 in SGC7901

  1. 犬弓首线虫感染小鼠脑组织细胞凋亡及凋亡基因bcl-2、bax的表达%Apoptosis in cerebral histiocytes and the expression of apoptosis-related genes Bcl-2 and bax in mice infected with Toxocara canis

    Institute of Scientific and Technical Information of China (English)

    郑胜生; 李建华; 沈继龙; 汪学龙

    2006-01-01

    目的研究犬弓首线虫(Toxocara canis)感染小鼠脑组织细胞凋亡及凋亡基因bcl-2、bax mRNA表达情况,探讨幼虫移行症对脑组织细胞影响可能机理.方法取狗小肠内犬弓首线虫成虫进行解剖,取子宫段的虫卵培养至感染期,感染小鼠后不同时间段取脑组织采用流式细胞仪检测小鼠脑组织细胞凋亡;应用原位杂交技术检测bcl-2、bax mRNA表达情况.结果 1.犬弓首线虫子宫段虫卵经培养有98%达感染期.2.病理切片HE染色均见感染小鼠脑组织有犬弓首线虫幼虫.3.流式细胞仪检测小鼠脑组织细胞凋亡10 d、15 d、20 d、25 d对照组与实验组比较,具统计学意义差别(P0.05).结论 1.犬弓首线虫感染小鼠早期,脑组织细胞有不同程度细胞凋亡出现.2.其细胞凋亡与凋亡基因bcl-2、bax无明显关系.

  2. Changes of bcl-xL and bax mRNA expression following traumatic brain injury in rats

    Institute of Scientific and Technical Information of China (English)

    骆纯; 卢亦成; 江基尧; 朱诚

    2002-01-01

    Objective: To investigate the changes of bcl-2 gene family and the molecular mechanism of neuronal apoptosis following traumatic brain injury (TBI) in rats.Methods: Male Sprague-Dawley (SD) rats were subjected to lateral fluid percussion brain injury (FPBI) of moderate severity. The bcl-xL and bax mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR). In addition to morphological evidence of apoptosis, terminal deoxynucleotide transferase-mediated dUTP-biotin nick-end labeling (TUNEL) histochemistry was used to identify the DNA fragmentation in situ at both light and electron microscope levels, whereas characteristic internucleosomal DNA fragmentation of apoptosis was demonstrated by DNA gel electrophoresis.Results: The apoptotic response to trauma was regionally distinct and may be involved in both acute and delayed cell death. The bcl-xL mRNA expression of the impact site was significantly lower (67.42%±7.54%) than that of the ipsilateral hemisphere at 6 hours after injury (P<0.01). The decrease of bcl-xL mRNA expression preceded apoptosis at 24 hours after injury. The bax mRNA expression rose slowly, doubled at 3 days after injury and returned to the sham level slowly.Conclusions: Decreased expression of bcl-xL mRNA and increased expression of bax mRNA coincides with apoptosis following brain injury. The bcl-2 gene family is involved in neuronal apoptosis after TBI, and the changes of mRNA expression of the family members lead the neuronal cells to apoptosis.

  3. A sandwich ELISA for the conformation-specific quantification of the activated form of human Bax.

    Science.gov (United States)

    Teijido, Oscar; Ganesan, Yogesh Tengarai; Llanos, Raul; Peton, Ashley; Urtecho, Jean-Baptiste; Soprani, Adauri; Villamayor, Aimee; Antonsson, Bruno; Manon, Stéphen; Dejean, Laurent

    2016-03-15

    Bcl-2 family proteins are critical regulators of mitochondrial outer membrane permeabilization (MOMP), which represents the point of no return of apoptotic cell death. The exposure of the Bax N-terminus at the mitochondria reflects Bax activation; and this activated configuration of the Bax protein is associated with MOMP. N-terminal exposure can be detected using specific monoclonal and/or polyclonal antibodies, and the onset of activated Bax has extensively been used as an early marker of apoptosis. The protocols of immunoprecipitation and/or immunocytochemistry commonly used to detect activated Bax are long and tedious, and allow semiquantification of the antigen at best. The sandwich ELISA protocol we developed has a 5 ng/mL detection limit and is highly specific for the activated conformation of Bax. This ELISA allows a rapid quantification of activated human Bax in whole cells and isolated mitochondria protein extracts. These properties grant this assay the potential to further clarify the prognostic and diagnostic value of activated Bax in disorders associated with deregulated apoptotic pathways such as degenerative diseases or cancer. PMID:26748144

  4. Involvement of nitric oxide signaling in mammalian Bax-induced terpenoid indole alkaloid production of Catharanthus roseus cells

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Bax, a mammalian pro-apoptotic member of the Bcl-2 family, has been demonstrated to be a potential regulatory factor for plant secondary metabolite biosynthesis recently. To investigate the molecular mechanism of Bax-induced secondary metabolite biosynthesis, we determined the contents of nitric oxide (NO) of the transgenic Catharanthus roseus cells overexpressing a mouse Bax protein and checked the effects of NO specific scavenger 2,4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1- oxyl-3-oxide (cPITO) on Bax-induced terpenoid indole alkaloid (TIA) production of the cells. The data showed that overexpression of the mouse Bax in C. roseus cells triggered NO generation of the cells. Treatment of cPITO not only inhibited the Bax-triggered NO burst but also suppressed the Bax-induced TIA production. The results indicated that the mouse Bax might activate the NO signaling in C. roseus cells and induce TIA production through the NO-dependent signal pathway in the cells. Furthermore, the activities of nitric oxide synthase (NOS) were significantly increased in the transgenic Bax cells as compared to those in the control cells, showing that the mouse Bax may induce NOS of C. roseus cells. Treatment of the transgenic Bax cells with NOS inhibitor PBITU blocked both Bax-induced NO generation and TIA production, which suggested that the mouse Bax might trigger NO generation and TIA production through NOS. However, the NOS-like activities and NO generation in the transgenic Bax cells did not match kinetically and the Bax-induced NOS-like activity was much later and lower than NO production. Moreover, the Bax-induced NO generation and TIA production were only partially inhibited by PBITU. Thus, our results suggested that the Bax-induced NO production and secondary metabolite biosynthesis in C. roseus cells was not entirely dependent on NOS or NOS-like enzymes.

  5. Overexpression of Bax sensitizes prostate cancer cells to TGF-β induced apoptosis

    Institute of Scientific and Technical Information of China (English)

    Pei Hui LIN; Zui PAN; Lin ZHENG; Na LI; David DANIELPOUR; Jian Jie MA

    2005-01-01

    NRP-154 is a tumorigenic epithelial cell line derived from the preneoplastic dorsal-lateral prostate of rats. These cells are exquisitely sensitive to TGF-β induced apoptosis. In contrast, we find that NRP-154 cells can sustain overexpression of exogenous Bax protein, which is different from non-tumor cells where Bax functions as a ubiquitous stimulator of apoptosis. NRP-154 cells stably overexpressing Bax show increased sensitivity to TGF-β induced apoptosis. The degree of TGF-β induced apoptosis displays high correlation with cleavage of Bax at the amino-terminus. Our data indicate that prostate cancer cells can host high levels of latent Bax which can be activated through post-translational modification.

  6. Molecular Basis for Membrane Pore Formation by Bax Protein Carboxyl Terminus

    Science.gov (United States)

    Tatulian, Suren A.; Garg, Pranav; Nemec, Kathleen N.; Chen, Bo; Khaled, Annette R.

    2015-01-01

    Bax protein plays a key role in mitochondrial membrane permeabilization and cytochrome c release upon apoptosis. Our recent data have indicated that the 20-residue C-terminal peptide of Bax (BaxC-KK; VTIFVAGVL-TASLTIWKKMG), when expressed intracellularly, translocates to the mitochondria and exerts lethal effect on cancer cells. Moreover, the BaxC-KK peptide, as well as two mutants where the two lysines are replaced with glutamate (BaxC-EE) or leucine (BaxC-LL), have been shown to form relatively large pores in lipid membranes, composed of up to eight peptide molecules per pore. Here the pore structure is analyzed by polarized Fourier transform infrared, circular dichroism, and fluorescence experiments on the peptides reconstituted in phospholipid membranes. The peptides assume an α/β-type secondary structure within membranes. Both β-strands and α-helices are significantly (by 30–60 deg) tilted relative to the membrane normal. The tryptophan residue embeds into zwitterionic membranes at 8–9 Å from the membrane center. The membrane anionic charge causes a deeper insertion of tryptophan for BaxC-KK and BaxC-LL but not for BaxC-EE. Combined with the pore stoichiometry determined earlier, these structural constraints allow construction of a model of the pore where eight peptide molecules form an “α/β-ring” structure within the membrane. These results identify a strong membranotropic activity of Bax C-terminus and propose a new mechanism by which peptides can efficiently perforate cell membranes. Knowledge on the pore forming mechanism of the peptide may facilitate development of peptide-based therapies to kill cancer or other detrimental cells such as bacteria or fungi. PMID:23110300

  7. 细胞凋亡及bax基因表达与舌苔变化关系的研究%A Study on the Relationship Between the Gene Expression of Bax and Cell Apoptosis in Tongue Mycoderma Epithelium of Common Tongue Coating

    Institute of Scientific and Technical Information of China (English)

    李新华; 吴正治; 李明; 张永锋; 陈嫚茵

    2006-01-01

    目的:检测常见舌苔舌上皮细胞凋亡情况及凋亡相关基因bax mRNA和蛋白产物,探讨舌苔厚度变化与舌上皮细胞凋亡、bax基因表达的关系.方法:运用TUNEL(末端脱氧核苷酸转移酶介导的脱氧尿嘧啶核苷三磷酸缺口末端标记)技术、原位杂交、免疫组化和图像分析技术.结果:与正常薄苔比较,剥苔bax基因过度表达伴随细胞凋亡增多,而厚苔bax基因低表达伴随细胞凋亡减少.bax基因表达水平变化趋势与细胞凋亡水平变化趋势一致.结论:bax基因表达水平的变化可能是影响舌苔上皮细胞凋亡并导致舌苔厚度变化的重要原因.

  8. PUMA promotes Bax translocation by competitive binding to Bcl-Xl during UV-induced apoptosis

    Science.gov (United States)

    Zhang, Yingjie; Xing, Da; Wu, Yinyuan; Liu, Lei

    2008-02-01

    Ultraviolet (UV) irradiation can induce apoptosis through both the membrane death receptor and the intrinsic apoptotic signaling pathways as DNA-damaging agents. PUMA, a BH3-only Bcl-2 family protein, plays an essential role in DNA damage-induced apoptosis. Bax, also a Bcl-2 family member, translocates from the cytosol to the mitochondrial membrane during UV-induced apoptosis. However, the regulation of Bax activation induced by UV irradiation remains poorly understood. In this study, the FRET (fluorescence resonance energy transfer) technique was used to study the interactions of Bax, Bcl-Xl, and PUMA in ASTC-a-1 cells. The results show that Bax translocated from the cytosol to the mitochondrial membrane at about 7 h after UV irradiation, and the translocation can not be blocked completely when overexpressed Bcl-xl. Moreover, The interaction of Bax and Bcl-Xl weakened markedly. In addition, Co-immunoprecipitation shows that PUMA released Bax by directly binding to Bcl-XL after UV irradiation in ASTC-a-1 cells. Taken together, these results indicated that PUMA can promote Bax translocation by binding to Bcl-Xl during UV-induced apoptosis.

  9. Insights into the structural stability of Bax from molecular dynamics simulations at high temperatures

    Science.gov (United States)

    Rosas-Trigueros, Jorge Luis; Correa-Basurto, José; Guadalupe Benítez-Cardoza, Claudia; Zamorano-Carrillo, Absalom

    2011-01-01

    Bax is a member of the Bcl-2 protein family that participates in mitochondrion-mediated apoptosis. In the early stages of the apoptotic pathway, this protein migrates from the cytosol to the outer mitochondrial membrane, where it is inserted and usually oligomerizes, making cytochrome c-compatible pores. Although several cellular and structural studies have been reported, a description of the stability of Bax at the molecular level remains elusive. This article reports molecular dynamics simulations of monomeric Bax at 300, 400, and 500 K, focusing on the most relevant structural changes and relating them to biological experimental results. Bax gradually loses its α-helices when it is submitted to high temperatures, yet it maintains its globular conformation. The resistance of Bax to adopt an extended conformation could be due to several interactions that were found to be responsible for maintaining the structural stability of this protein. Among these interactions, we found salt bridges, hydrophobic interactions, and hydrogen bonds. Remarkably, salt bridges were the most relevant to prevent the elongation of the structure. In addition, the analysis of our results suggests which conformational movements are implicated in the activation/oligomerization of Bax. This atomistic description might have important implications for understanding the functionality and stability of Bax in vitro as well as within the cellular environment. PMID:21936009

  10. Bax-PGAM5L-Drp1 complex is required for intrinsic apoptosis execution.

    Science.gov (United States)

    Xu, Wenjuan; Jing, Linlin; Wang, Quanshi; Lin, Chung-Chih; Chen, Xiaoting; Diao, Jianxin; Liu, Yuanliang; Sun, Xuegang

    2015-10-01

    Intrinsic apoptosis eliminates cells with damaged DNA and cells with dysregulated expression of oncogene. PGAM5, a member of the phosphoglycerate mutase family, has two splicing variants: PGAM5L (the long form) and PGAM5S (the short form). It has been well established that PGAM5 is at the convergent point of multiple necrosis pathways. However, the role of PGAM5 in intrinsic apoptosis is still controversial. Here we report that the PGAM5L, but not PGAM5S is a prerequisite for the activation of Bax and dephosphorylation of Drp1 in arenobufagin and staurosporine induced intrinsic apoptosis. Knockdown of PGAM5L inhibits the translocation of Bax to the mitochondria and reduces mitochondrial fission. The interaction between PGAM5L and Drp1 was observed in both arenobufagin and staurosporine treated HCT116 cells, but not in HCT116 Bax(-/-) cells. Bax transfection rescues the formation of the triplex in both arenobufagin and staurosporine stimulated HCT116 Bax(-/-) cells. Arenobufagin shows remarkable anti-cancer effects both in orthotropic and heterotropic CRC models and demonstrates less toxic effects as compared with that of cisplatin. Bax-PGAM5L-Drp1 complex is detected in arenobufagin and staurosporine treated CRC cells in vitro and in arenobufagin and cisplatin treated tumor in vivo as well. In summary, our results demonstrate that Bax-PGAM5L-Drp1 complex is required for intrinsic apoptosis execution. PMID:26356820

  11. Isatin decreases Bax protein expression in the substantia nigra of a mouse model of Parkinson's disease

    Institute of Scientific and Technical Information of China (English)

    Jiguo Zhang; Fang Zhang; Yanlong Qiu; Wang Yue

    2011-01-01

    The present study observed the action of 1H-indole-2, 3-dione (isatin) on Bax protein expression in the substantia nigra of a Parkinson's disease animal model. Parkinson's disease-like behaviors were induced in C57BL/6J mice treated with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Bax protein expression was significantly reduced in isatin (100, 200 mg/kg)-pretreated mice. Results demonstrate that isatin plays a neuroprotective role in mice treated with MPTP by down-regulating Bax protein expression.

  12. Bax translocation into mitochondria during dihydroartemisinin(DHA)-induced apoptosis in human lung adenocarcinoma cells

    Science.gov (United States)

    Lu, Ying-ying; Chen, Tong-sheng; Qu, Jun-Le

    2009-02-01

    Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, isolated from the traditional Chinese herb Artemisia annua, has been shown to possess promising anticancer activities and induce cancer cell death through apoptotic pathways. However, the molecular mechanisms are not well understood. This study was investigated in human lung adenocarconoma ASTC-a-1 cell line and aimed to determine whether the apoptotic process was mediated by Bax activation and translocation during DHA-induced apoptosis. In this study, DHA induced a time-dependent apoptotic cell death, which was assayed by Cell Counting Kit (CCK-8) and Hoechst 33258 staining. Detection of Bax aggregation and translocation to mitochondria was observed in living cells which were co-transfected with GFP-Bax and Dsred-mito plasmid using confocal fluorescence microscope technique. Overall, these results demonstrated that Bax activation and translocation to mitochondria occurred during DHA-induced apoptosis.

  13. Paclitaxel induces apoptosis in human gastric carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    Hai-Bo Zhou; Ju-Ren Zhu

    2003-01-01

    AIM: To investigate the apoptosis in gastric cancer cells induced by paclitaxel, and the relation between this apoptosis and expression of Bcl-2 and Bax.METHODS: In in vitro experiments, MTT assay was used to determine the cell growth inhibitory rate. Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of gastric cancer cell line SGC-7901 before and after the paditaxel treatment. Immunohistochemical staining was used to detect the expression of apoptosis-regulated gene Bcl-2and Bax.RESULTS: Paclitaxel inhibited the growth of gastric cancer cell line SGC-7901 in a dose-and time-dependent manner.Paclitaxel induced SGC-7901 cells to undergo apoptosis with typically apoptotic characteristics, including morphological changes of chromatin condensation, chromatin crescent formation, nucleus fragmentation and apoptotic body formation. Paclitaxel could reduce the expression of apoptosis-regulated gene Bcl-2, and improve the expression of apoptosis-regulated gene Bax.CONCLUSION: Paclitaxel is able to induce the apoptosis in gastric cancer. This apoptosis may be mediated by downexpression of apoptosis-regulated gene Bcl-2 and upexpression of apoptosis-regulated gene Bax.

  14. Expression of Arabidopsis Bax Inhibitor-1 in transgenic sugarcane confers drought tolerance.

    Science.gov (United States)

    Ramiro, Daniel Alves; Melotto-Passarin, Danila Montewka; Barbosa, Mariana de Almeida; Santos, Flavio Dos; Gomez, Sergio Gregorio Perez; Massola Júnior, Nelson Sidnei; Lam, Eric; Carrer, Helaine

    2016-09-01

    The sustainability of global crop production is critically dependent on improving tolerance of crop plants to various types of environmental stress. Thus, identification of genes that confer stress tolerance in crops has become a top priority especially in view of expected changes in global climatic patterns. Drought stress is one of the abiotic stresses that can result in dramatic loss of crop productivity. In this work, we show that transgenic expression of a highly conserved cell death suppressor, Bax Inhibitor-1 from Arabidopsis thaliana (AtBI-1), can confer increased tolerance of sugarcane plants to long-term (>20 days) water stress conditions. This robust trait is correlated with an increased tolerance of the transgenic sugarcane plants, especially in the roots, to induction of endoplasmic reticulum (ER) stress by the protein glycosylation inhibitor tunicamycin. Our findings suggest that suppression of ER stress in C4 grasses, which include important crops such as sorghum and maize, can be an effective means of conferring improved tolerance to long-term water deficit. This result could potentially lead to improved resilience and yield of major crops in the world. PMID:26872943

  15. Lack of association between Bax promoter (-248G>A single nucleotide polymorphism and susceptibility towards cancer: evidence from a meta-analysis.

    Directory of Open Access Journals (Sweden)

    Sushil Kumar Sahu

    Full Text Available BACKGROUND: The Bcl-2-associated X protein (Bax is a proapoptotic member of the Bcl-2 family known to be activated and upregulated during apoptosis. Single nucleotide polymorphisms (SNPs in Bax promoter may participate in the process of carcinogenesis by altering its own expression and the cancer related genes. Bax-248G>A polymorphism has been implicated to alter the risk of cancer, but the listed results are inconsistent and inconclusive. In the present study, we performed a meta-analysis to systematically summarize the possible association of this polymorphism with the risk of cancer. METHODOLOGY: We conducted a search of case-control studies on the associations of Bax-248G>A polymorphism with susceptibility to cancer in Pub Med, Science Direct, Wiley Online Library and hand search. Data from all eligible studies based on some key search terms, inclusion and exclusion criteria were extracted for this meta-analysis. Hardy-Weinberg equilibrium (HWE in controls, power calculation, heterogeneity analysis, Begg's funnel plot, Egger's linear regression test, forest plot and sensitivity analysis were performed in the present study. RESULTS: Cancer risk associated with Bax-248G>A polymorphism was estimated by pooled odds ratios (ORs and 95% confidence intervals (95% CIs. The pooled ORs were calculated in allele contrast, homozygous comparison, heterozygous comparison, dominant and recessive model. Statistical significance was checked through Z and p-value in forest plot. A total of seven independent studies including 1772 cases and 1708 controls were included in our meta-analysis. Our results showed that neither allele frequency nor genotype distributions of this polymorphism were associated with risk for cancer in any of the genetic model. Furthermore, Egger's test did not show any substantial evidence of publication bias. CONCLUSIONS/SIGNIFICANCE: This meta-analysis suggests that the Bax-248G>A polymorphism is not an important cancer risk factor

  16. A New Fungal Diterpene Induces VDAC1-dependent Apoptosis in Bax/Bak-deficient Cells.

    Science.gov (United States)

    Huang, Li; Han, Junjie; Ben-Hail, Danya; He, Luwei; Li, Baowei; Chen, Ziheng; Wang, Yueying; Yang, Yanlei; Liu, Lei; Zhu, Yushan; Shoshan-Barmatz, Varda; Liu, Hongwei; Chen, Quan

    2015-09-25

    The pro-apoptotic Bax and Bak proteins are considered central to apoptosis, yet apoptosis occurs in their absence. Here, we asked whether the mitochondrial protein VDAC1 mediates apoptosis independently of Bax/Bak. Upon screening a fungal secondary metabolite library for compounds inducing apoptosis in Bax/Bak-deficient mouse embryonic fibroblasts, we identified cyathin-R, a new cyathane diterpenoid compound able to activate apoptosis in the absence of Bax/Bak via promotion of the VDAC1 oligomerization that mediates cytochrome c release. Diphenylamine-2-carboxilic acid, an inhibitor of VDAC1 conductance and oligomerization, inhibited cyathin-R-induced VDAC1 oligomerization and apoptosis. Similarly, Bcl-2 overexpression conferred resistance to cyathin-R-induced apoptosis and VDAC1 oligomerization. Silencing of VDAC1 expression prevented cyathin-R-induced apoptosis. Finally, cyathin-R effectively attenuated tumor growth and induced apoptosis in Bax/Bak-deficient cells implanted into a xenograft mouse model. Hence, this study identified a new compound promoting VDAC1-dependent apoptosis as a potential therapeutic option for cancerous cells lacking or presenting inactivated Bax/Bak.

  17. A New Fungal Diterpene Induces VDAC1-dependent Apoptosis in Bax/Bak-deficient Cells*

    Science.gov (United States)

    Huang, Li; Han, Junjie; Ben-Hail, Danya; He, Luwei; Li, Baowei; Chen, Ziheng; Wang, Yueying; Yang, Yanlei; Liu, Lei; Zhu, Yushan; Shoshan-Barmatz, Varda; Liu, Hongwei; Chen, Quan

    2015-01-01

    The pro-apoptotic Bax and Bak proteins are considered central to apoptosis, yet apoptosis occurs in their absence. Here, we asked whether the mitochondrial protein VDAC1 mediates apoptosis independently of Bax/Bak. Upon screening a fungal secondary metabolite library for compounds inducing apoptosis in Bax/Bak-deficient mouse embryonic fibroblasts, we identified cyathin-R, a new cyathane diterpenoid compound able to activate apoptosis in the absence of Bax/Bak via promotion of the VDAC1 oligomerization that mediates cytochrome c release. Diphenylamine-2-carboxilic acid, an inhibitor of VDAC1 conductance and oligomerization, inhibited cyathin-R-induced VDAC1 oligomerization and apoptosis. Similarly, Bcl-2 overexpression conferred resistance to cyathin-R-induced apoptosis and VDAC1 oligomerization. Silencing of VDAC1 expression prevented cyathin-R-induced apoptosis. Finally, cyathin-R effectively attenuated tumor growth and induced apoptosis in Bax/Bak-deficient cells implanted into a xenograft mouse model. Hence, this study identified a new compound promoting VDAC1-dependent apoptosis as a potential therapeutic option for cancerous cells lacking or presenting inactivated Bax/Bak. PMID:26253170

  18. Bcl-xS and Bax induce different apoptotic pathways in PC12 cells.

    Science.gov (United States)

    Lindenboim, L; Yuan, J; Stein, R

    2000-03-30

    Apoptosis is regulated by the action of the Bcl-2 family of proteins, which includes anti- and pro-apoptotic members such as Bcl-xS and Bax. These proteins may differ from each other in structure, mechanism of action and interactions with anti-apoptotic signaling. The mechanism whereby Bax induces cell death has been studied in some cellular systems, but the mechanism of Bcl-xS-induced apoptosis is largely unknown. In this study we investigated and compared the apoptotic effects of Bcl-xS and Bax in the pheochromocytoma cell line, PC12 (a useful model system for studying neuronal apoptosis), and the extent to which they are protected by the survival factor, nerve growth factor (NGF). PC12 cells express endogenous Bcl-xS, Bax and Bcl-xL proteins. Subcellular fractionation revealed that Bax is presented mainly in the cytosolic and the heavy membrane fractions, Bcl-xS is present only in the cytosol, and the anti-apoptotic protein Bcl-xL is located mainly in the heavy membrane fraction. In contrast to the cytosolic localization of endogenous Bcl-xS, the exogenously overexpressed Bcl-xS is localized to the mitochondria. Overexpression of Bcl-xS or Bax induces cell death in the transfected cells. The cell death induced by overexpression of Bcl-xS was inhibited by coexpression of Bcl-xS with Bcl-2 or Bcl-xL, or by treatment with the broad-spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoro-methylketone (Z-VAD-FMK) or with NGF. The Bcl-2 mutants deltaC22, which lacks the transmembrane domain, and G145A (mI-3) were able to inhibit the death-inducing effect of Bcl-xS. These results therefore suggest that the apoptotic pathway induced by overexpression of Bcl-xS in PC12 cells can be controlled by Bcl-2 and Bcl-xL, is mediated by caspases, and can be inhibited by the NGF signaling pathway. The Bax-induced cell death was inhibited by co-expression of Bax with Bcl-2 or Bcl-xL, but was not inhibited by Z-VAD-FMK, NGF, or the Bcl-2 ml-3 or deltaC22 mutants. These

  19. Dynamic interaction between 14-3-3zeta and bax during TNF-α-induced apoptosis in living cells

    Science.gov (United States)

    Gao, Xuejuan; Xing, Da; Chen, Tongsheng

    2006-09-01

    Bax, a proapoptotic member of the Bcl-2 family, localizes largely in the cytoplasm but redistributes to mitochondria and undergoes oligomerization to induce the release of apoptogenic factors such as cytochrome c in response to apoptotic stimuli. Cytoplasmic protein 14-3-3zeta binds to Bax and, upon apoptotic stimulation, releases Bax by a caspase-independent mechanism. However, the direct interaction of the cytoplasmic 14-3-3zeta and Bax in living cells has not been observed. In present study, to monitor the dynamic interaction between 14-3-3zeta and Bax in living cells in real time during apoptosis induced by tumor necrosis factor (TNF-α), DsRed-14-3-3zeta plasmid is constructed. By cotransfecting DsRed- 14-3-3zeta and GFP-Bax plasmids into human lung adenocarcinoma cells (ASTC-a-1), we observe the dynamic interaction between Bax and 14-3-3zeta using fluorescence resonance energy transfer (FRET) technique on laser scanning confocal microscope. The results show that 14-3-3zeta remains in the cytoplasm but GFP-Bax translocates to mitochondria completely after TNF-α stimulation. These results reveal that 14-3-3zeta binds directly to Bax in healthy cells, and that 14-3-3zeta negatively regulates Bax translocation to mitochondria during TNF-α-induced apoptosis.

  20. Both p53-PUMA/NOXA-Bax-mitochondrion and p53-p21cip1 pathways are involved in the CDglyTK-mediated tumor cell suppression

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Zhendong, E-mail: zdyu@hotmail.com [Department of Clinical laboratory, Peking University Shenzhen Hospital, Guangdong (China); Wang, Hao [Department of pathology, The Chinese University of Hong Kong, Hong Kong (China); Zhang, Libin; Tang, Aifa; Zhai, Qinna; Wen, Jianxiang; Yao, Li [Department of Clinical laboratory, Peking University Shenzhen Hospital, Guangdong (China); Li, Pengfei, E-mail: lipengfei@cuhk.edu.hk [Department of pathology, The Chinese University of Hong Kong, Hong Kong (China)

    2009-09-04

    CDglyTK fusion suicide gene has been well characterized to effectively kill tumor cells. However, the exact mechanism and downstream target genes are not fully understood. In our study, we found that CDglyTK/prodrug treatment works more efficiently in p53 wild-type (HONE1) cells than in p53 mutant (CNE1) cells. We then used adenovirus-mediated gene delivery system to either knockdown or overexpress p53 and its target genes in these cells. Consistent results showed that both p53-PUMA/NOXA/Bcl2-Bax and p53-p21 pathways contribute to the CDglyTK induced tumor cell suppression. Our work for the first time addressed the role of p53 related genes in the CDglyTK/prodrug system.

  1. Both p53-PUMA/NOXA-Bax-mitochondrion and p53-p21cip1 pathways are involved in the CDglyTK-mediated tumor cell suppression

    International Nuclear Information System (INIS)

    CDglyTK fusion suicide gene has been well characterized to effectively kill tumor cells. However, the exact mechanism and downstream target genes are not fully understood. In our study, we found that CDglyTK/prodrug treatment works more efficiently in p53 wild-type (HONE1) cells than in p53 mutant (CNE1) cells. We then used adenovirus-mediated gene delivery system to either knockdown or overexpress p53 and its target genes in these cells. Consistent results showed that both p53-PUMA/NOXA/Bcl2-Bax and p53-p21 pathways contribute to the CDglyTK induced tumor cell suppression. Our work for the first time addressed the role of p53 related genes in the CDglyTK/prodrug system.

  2. Changes of bcl—XL and bax mRNA expression following traumatic brain injury in rats

    Institute of Scientific and Technical Information of China (English)

    骆纯; 卢亦成; 等

    2002-01-01

    Objective:To investigate the changes of bcl-2 gene family and the molecular mechanism of neuromal apoptosis following traumatic brain injury(TBI)in rats.Methods:Male Sprague-Dawley(SD)rats were subjected to lateral fluid percussion brain injury(FPBI)of moderate severity.Thebcl-XLand baxmRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR)bax mRNA expression rose slowly,doubled at 3days after injury and returned to the sham level slowly.Conclusions:Decreased expression of bcl-XLmRNA and increased expression of bax mRNA coincides tith apoptosis followwin brain injury.The bcl-2gene family is involved in neuronal apoptosis after TBI,and the changes of mRNA expression of the family members lead the neuronal cells to apoptosis.

  3. Identification of Bax-interacting proteins in oligodendrocyte progenitors during glutamate excitotoxicity and perinatal hypoxia–ischemia

    Directory of Open Access Journals (Sweden)

    Sopio Simonishvili

    2013-12-01

    Full Text Available OPC (oligodendrocyte progenitor cell death contributes significantly to the pathology and functional deficits following hypoxic-ischemic injury in the immature brain and to deficits resulting from demyelinating diseases, trauma and degenerative disorders in the adult CNS. Glutamate toxicity is a major cause of oligodendroglial death in diverse CNS disorders, and previous studies have demonstrated that AMPA/kainate receptors require the pro-apoptotic protein Bax in OPCs undergoing apoptosis. The goal of the present study was to define the pro-apoptotic and anti-apoptotic effectors that regulate Bax in healthy OPCs and after exposure to excess glutamate in vitro and following H–I (hypoxia–ischemia in the immature rat brain. We show that Bax associates with a truncated form of Bid, a BH3-only domain protein, subsequent to glutamate treatment. Furthermore, glutamate exposure reduces Bax association with the anti-apoptotic Bcl family member, Bcl-xL. Cell fractionation studies demonstrated that both Bax and Bid translocate from the cytoplasm to mitochondria during the early stages of cell death consistent with a role for Bid as an activator, whereas Bcl-xL, which normally complexes with both Bax and Bid, disassociates from these complexes when OPCs are exposed to excess glutamate. Bax remained unactivated in the presence of insulin-like growth factor-1, and the Bcl-xL complexes were protected. Our data similarly demonstrate loss of Bcl-xL–Bax association in white matter following H–I and implicate active Bad in Bax-mediated OPC death. To identify other Bax-binding partners, we used proteomics and identified cofilin as a Bax-associated protein in OPCs. Cofilin and Bax associated in healthy OPCs, whereas the Bax–cofilin association was disrupted during glutamate-induced OPC apoptosis.

  4. Apoptosis of human tumor cells by chemotherapeutic anthracyclines is enhanced by Bax overexpression.

    Science.gov (United States)

    Lu, Y; Yagi, T

    1999-09-01

    One of the major factors for efficacy of a chemotherapeutic drug is its activity to induce apoptosis of tumor cells. Doxorubicin and daunorubicin, radiomimetic anthracycline-group drugs, have been used for chemotherapy for about 30 years. Here we established the colorectal tumor and osteosarcoma cells in which Bax expression can be induced by the treatment of isopropyl-beta-D-thiogalactopyranoside, and examined the effect of the Bax overexpression on the cell death caused by these drugs. While the Bax overexpression neither affected growth nor morphology of the undamaged cells, it enhanced the cell death caused by these drugs. Increase in cellular nucleus fragmentation and DNA ladder formation indicates that the Bax-enhanced cell death is due to enhanced apoptosis of the drug-treated cells. The enhanced cell death was not observed when the cells were irradiated with X-ray or treated with other chemotherapeutic agents we examined. These results indicate that Bax may have a specific role to enhance the efficacy of chemotherapy with anthracycline-group agents.

  5. Eucommia ulmoides cortex, geniposide and aucubin regulate lipotoxicity through the inhibition of lysosomal BAX.

    Directory of Open Access Journals (Sweden)

    Geum-Hwa Lee

    Full Text Available In this study we examined the inhibition of hepatic dyslipidemia by Eucommia ulmoides extract (EUE. Using a screening assay for BAX inhibition we determined that EUE regulates BAX-induced cell death. Among various cell death stimuli tested EUE regulated palmitate-induced cell death, which involves lysosomal BAX translocation. EUE rescued palmitate-induced inhibition of lysosomal V-ATPase, α-galactosidase, α-mannosidase, and acid phosphatase, and this effect was reversed by bafilomycin, a lysosomal V-ATPase inhibitor. The active components of EUE, aucubin and geniposide, showed similar inhibition of palmitate-induced cell death to that of EUE through enhancement of lysosome activity. Consistent with these in vitro findings, EUE inhibited the dyslipidemic condition in a high-fat diet animal model by regulating the lysosomal localization of BAX. This study demonstrates that EUE regulates lipotoxicity through a novel mechanism of enhanced lysosomal activity leading to the regulation of lysosomal BAX activation and cell death. Our findings further indicate that geniposide and aucubin, active components of EUE, may be therapeutic candidates for non-alcoholic fatty liver disease.

  6. Bax is not involved in the resveratrol-induced apoptosis in human lung adenocarcinoma cells

    Science.gov (United States)

    Zhang, Wei-wei; Wang, Zhi-ping; Chen, Tong-sheng

    2010-02-01

    Resveratrol (RV) is a natural plant polyphenol widely present in foods such as grapes, wine, and peanuts. Previous studies indicate that RV has an ability to inhibit various stages of carcinogenesis and eliminate preneoplastic cells in vitro and in vivo. However, little is known about the molecular mechanism of RV-induced apoptosis in human lung adenocarcinoma (ASTC-a-1) cell. In this report, we analyzed whether Bax translocation from cytoplasm to mitochondria during RV-induced apoptosis in single living cell using onfocal microscopey. Cells were transfected with GFP-Bax plasmid. Cell counting kit (CCK-8) assay was used to assess the inhibition of RV on the cells viability. Apoptotic activity of RV was detected by Hoechst 33258 and propidium iodide (PI) staining. Our results showed that RV induced a dose-dependent apoptosis in which Bax did not translocate to mitochondrias.

  7. Curcumin induces the expression of NF-κB and Bcl-2/Bax in human renal cell carcinoma cell line ACHN

    Institute of Scientific and Technical Information of China (English)

    Gang Li; Tie Chong; Ziming Wang

    2009-01-01

    Objective: To explore the in vitro effects of curcumin on the proliferation and apoptosis of the human renal cell carcinoma cell line ACHN, and to investigate its mechanisms of action. Methods: The human renal cell carcinoma cell line ACHN was treated with different concentrations of curcumin for 24 h. The MTT assay was used to evaluate the cytotoxic effects of curcumin and flow cytometry was utilized to observe and detect the apoptosis of ACHN cells induced by curcumin. The expression levels of Bcl-2, Bax and NF-κBP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), while the expression of Bcl-2, Bax, NF-κBP65 and IkB proteins was evaluated by Western blot. Results: The concentrations of curcumin used significantly inhibited the proliferation of ACHN human renal cell carcinoma cells in vitro in a dose and time-dependent manner (Ftime=5.55, P < 0.05; Fdose=110.05, P < 0.05). Obvious apoptosis of cells treated with different concentrations of curcumin could be observed by FCM. Compared with the control group, the apoptosis rates of curcumin-treated cells were markedly increased (F=96.35, P < 0.05). Lower dose of curcumin significantly induced the apoptosis of ACHN cells. With intervention of different concentrations of curcumin (0, 10, 20 and 40 μmol/L) for 24 h, the expression levels of Bcl-2 and NF-κBP65 mRNA in ACHN cells were decreased while the expression level of Bax mRNA was increased (P < 0.05), and Bcl-2, and NF-κBP65 protein decreased, while Bax and IκB protein increased compared with those in the untreated group. Conclusion: Curcumin inhibited proliferation and increased apoptosis of the human renal cell carcinoma cell line ACHN. These curcumin effects appear to involve up-regulating IκB, down-regulating NF-κB, and regulating the expression of the apoptosis genes Bcl-2/Bax.

  8. PUMA Promotes Bax Translocation by Both Directly Interacting with Bax and by Competitive Binding to Bcl-XL during UV-induced Apoptosis

    OpenAIRE

    Zhang, Yingjie; Xing, Da; Liu, Lei

    2009-01-01

    Cell apoptosis induced by UV irradiation is a highly complex process in which different molecular signaling pathways are involved. p53 up-regulated modulator of apoptosis (PUMA) has been proposed as an important regulator in UV irradiation-induced apoptosis. However, the molecular mechanism through which PUMA regulates apoptosis, especially how PUMA activates Bcl-2-associated X protein (Bax) in response to UV irradiation is still controversial. In this study, by using real-time single-cell an...

  9. Differential expression of Bcl-2 and Bax during gastric ischemia-reperfusion of rats

    Institute of Scientific and Technical Information of China (English)

    Wei-Li Qiao; Guang-Ming Wang; Yue Shi; Jin-Xia Wu; You-Jian Qi; Jian-Fu Zhang; Hong Sun; Chang-Dong Yan

    2011-01-01

    AIM: To investigate expression of Bcl-2 and Bax in gastric ischemia-reperfusion (GI-R) and involvement of extracellular signal-regulated kinase (ERK) 1/2 activation.METHODS: The GI-R model was established by ligature of the celiac artery for 30 min and reperfusion in Sprague-Dawley rats. Rats were assigned to groups in accordancewith their evaluation period: control, 0, 0.5, 1, 3, 6, 24,48, and 72 h. Expression and distribution of Bcl-2 and Bax proteins were analyzed by immunohistochemistry and western blotting in gastric tissue samples after sacrifice.RESULTS: Compared with controls, the percentage of positive cells and protein levels of Bcl-2 decreased in the early phases of reperfusion, reached its minimumat 1 h (P < 0.05); it then increased, reaching its peak at 24 h of reperfusion (P < 0.05). The pattern of Bax expression was opposite to that of Bcl-2. Bax expressionincreased after reperfusion, with its peak at 1 h of reperfusion (P < 0.05), and then it decreased gradually to a minimum at 24 h after reperfusion (P < 0.05).On the other hand, inhibition of activation of ERK1/2 induced by PD98059, a specific upstream MEK inhibitor,had significant effects on Bcl-2 and Bax in GI-R.Compared with GI-R treatment only at 3 h of reperfusion,PD98059 reduced the number of Bcl-2 positive cells (0.58% of R3h group, P < 0.05) and Bcl-2 proteinlevel (74% of R3h group, P < 0.05) but increased the number of Bax-positive cells (1.33-fold vs R3h group, P< 0.05) and Bax protein level (1.35-fold of R3h group,P < 0.05).CONCLUSION: These results indicated that the Bcl-2 and Bax played a pivotal role in the gastric mucosal I-R injury and repair by activation of ERK1/2.

  10. 杏仁核点燃鼠海马不同亚区Bax mRNA的表达%The expression of Bax mRNA in the hippocampus-subareas in amygdala-kindled rats

    Institute of Scientific and Technical Information of China (English)

    梁代义; 伍国锋; 庞成

    2006-01-01

    目的:观察细胞凋亡调控基因Bax mRNA在癫(癎)鼠海马不同亚区的表达.方法:利用电极植入鼠脑杏仁核的方法建立点燃癫(癎)模型,采用原位杂交法检测鼠脑海马CA1、CA2、CA3及齿状回(DGL)区Bax mRNA的表达.结果:正常大鼠海马各区少见Bax mRNA阳性细胞表达,杏仁核植入电极大鼠及其点燃后海马各区Bax mRNA表达阳性细胞数增多,点燃鼠Bax mRNA阳性细胞平均光密度高于仅仅植入电极鼠,海马不同亚区Bax mRNA阳性细胞平均光密度值不同.结论:杏仁核点燃癫(癎)模型鼠的海马不同亚区均存在Bax mRNA表达增强,但不同亚区对Bax mRNA表达敏感性不同,以DGL区为最高.

  11. Expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax after delayed paraplegia induced by ischemia/reperfusion injury

    Institute of Scientific and Technical Information of China (English)

    Bibo Liu; Miao Liu; Duoning Wang; Wei Ma; Shengli Dang

    2006-01-01

    BACKGROUND:Operation of spine does not involve in spinal cord;however,spinal cord injury occurs at post-operation induced by unclear factors.Meanwhile,after decompression of lumbar spinal canal,symptoms are severer and severer.In addition,during extirpation of oervical and lumbar intervertabral disc,spinal cord and its vessels are not damaged,but spinal cord injury is also suffered from patients with partial improvement.All statuses mentioned above are related to expressed changes of bc/-2 gene inhibiting apoptosis and bax gene accelerating apoptosis.OBJECTIVE:To observe motor function of hindlimbs of ischemia/reperfusion model in rabbits at various time points after reperfusion and expressions of apoptosis correlated protein Bcl-2 and Bax.DESIGN:Completely randomized grouping design and contrast study.SEITING:Department of Orthopedics,the First Affiliated Hospital of Medical School,Xi'an Jiao Tong University.MATERIALS:Forty-eight New Zealand white rabbits of both genders were randomly divided into sham operation group(n=24) and model group(n=24),and then,rabbits in each group were observed at four time points:8,24,72 and 168 hours after reperfusion,with 6 in each time point.Rabbit-anti-rabbit Bcl-2 antibody and rabbit-anti-rabbit Bax antibody were provided by Boster Company.The procedures were accordant to the METHODS:The experiment was carried out at Laboratory of Orthopaedics of First Affiliated Hospital of Medical School,Xi'an Jiao Tong University from April to August 2005.①Delayed paralysis models of spinal cord after ischemia/reperfusion were established based on method of Zivin et al.Animals in sham operation group underwent an exposure of abdominal aorta but the aorta was not occluded.②Motor function of hindlimb was observed 8,24,72 and 168 hours after reperfusion.A grade of 0-5 was assigned to each animal (grade 0:no voluntary hind limb function;grade 5:normal hop;grades 0-3:paraplegia).③The lumbar segment of the spinal cord(L3 to L5)was used for

  12. NDV-induced apoptosis in absence of Bax; evidence of involvement of apoptotic proteins upstream of mitochondria

    Directory of Open Access Journals (Sweden)

    Molouki Aidin

    2012-08-01

    Full Text Available Abstract Background Recently it was shown that following infection of HeLa cells with Newcastle disease virus (NDV, the matrix (M protein binds to Bax and subsequently the intrinsic pathway of apoptosis is activated. Moreover, there was very little alteration on mRNA and protein levels of Bax and Bcl-2 after infection with NDV. Finding In order to further investigate the role of members of the Bcl-2 family, Bax-knockout and wild-type HCT116 cells were infected with NDV strain AF2240. Although both cells underwent apoptosis through the activation of the intrinsic pathway and the release of cytochrome c from mitochondria, the percentage of dead Bax-knockout cells was significantly lower than wt cells (more than 10% at 48 h post-infection. In a parallel experiment, the effect of NDV on HT29 cells, that are originally Bcl-2-free, was studied. Apoptosis in HT29 cells was associated with Bax redistribution from cytoplasm to mitochondria, similar to that of HeLa and wt HCT116 cells. Conclusion Although the presence of Bax during NDV-induced apoptosis contributes to a faster cell death, it was concluded that other apoptotic protein(s upstream of mitochondria are also involved since cancer cells die whether in the presence or absence of Bax. Therefore, the classic Bax/Bcl-2 ratio may not be a major determinant in NDV-induced apoptosis.

  13. A BAX/BAK and cyclophilin D-independent intrinsic apoptosis pathway.

    Directory of Open Access Journals (Sweden)

    Sebastián Zamorano

    Full Text Available Most intrinsic death signals converge into the activation of pro-apoptotic BCL-2 family members BAX and BAK at the mitochondria, resulting in the release of cytochrome c and apoptosome activation. Chronic endoplasmic reticulum (ER stress leads to apoptosis through the upregulation of a subset of pro-apoptotic BH3-only proteins, activating BAX and BAK at the mitochondria. Here we provide evidence indicating that the full resistance of BAX and BAK double deficient (DKO cells to ER stress is reverted by stimulation in combination with mild serum withdrawal. Cell death under these conditions was characterized by the appearance of classical apoptosis markers, caspase-9 activation, release of cytochrome c, and was inhibited by knocking down caspase-9, but insensitive to BCL-X(L overexpression. Similarly, the resistance of BIM and PUMA double deficient cells to ER stress was reverted by mild serum withdrawal. Surprisingly, BAX/BAK-independent cell death did not require Cyclophilin D (CypD expression, an important regulator of the mitochondrial permeability transition pore. Our results suggest the existence of an alternative intrinsic apoptosis pathway emerging from a cross talk between the ER and the mitochondria.

  14. Zerumbone induced apoptosis in liver cancer cells via modulation of Bax/Bcl-2 ratio

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    Azimahtol Hawariah LP

    2007-04-01

    Full Text Available Abstract Background Zerumbone is a cytotoxic component isolated from Zingiber zerumbet Smith, a herbal plant which is also known as lempoyang. This new anticancer bioactive compound from Z. zerumbet was investigated for its activity and mechanism in human liver cancer cell lines. Results Zerumbone significantly showed an antiproliferative activity upon HepG2 cells with an IC50 of 3.45 ± 0.026 μg/ml. Zerumbone was also found to inhibit the proliferation of non-malignant Chang Liver and MDBK cell lines. However the IC50 obtained was higher compared to the IC50 for HepG2 cells (> 10 μg/ml. The extent of DNA fragmentation was evaluated by the Tdt-mediated dUTP nick end labelling assay which showed that, zerumbone significantly increased apoptosis in HepG2 cells in a time-course manner. In detail, the apoptotic process triggered by zerumbone involved the up-regulation of pro-apoptotic Bax protein and the suppression of anti-apoptotic Bcl-2 protein expression. The changes that occurred in the levels of this antagonistic proteins Bax/Bcl-2, was independent of p53 since zerumbone did not affect the levels of p53 although this protein exists in a functional form. Western blotting analysis for Bax protein was further confirmed qualitatively with an immunoassay that showed the distribution of Bax protein in zerumbone-treated cells. Conclusion Therefore, zerumbone was found to induce the apoptotic process in HepG2 cells through the up and down regulation of Bax/Bcl-2 protein independently of functional p53 activity.

  15. IMPORTANCE OF APOPTOSIS MARKERS (MDM2, BCL-2 AND Bax) IN CONVENTIONAL RENAL CELL CARCINOMA.

    Science.gov (United States)

    Saker, Z; Tsintsadze, O; Jiqia, I; Managadze, L; Chkhotua, A

    2015-12-01

    The goal of the current study was to analyze the expression of Bcl-2, MDM2 and Bax in benign and malignant renal tissue samples and assess their possible association with different clinical parameters. Prognostic significance of the markers in recurrence-free and cancer-specific survivals has also been evaluated. Activity of MDM2, Bcl-2 and Bax was evaluated in: 24 normal human kidney tissues resected from the patients of different ages (range: 21-80 years), and in 52 conventional RCC samples. Intensity of the markers' expression was compared between the groups and correlation was analyzed with different clinical parameters. Activity of anti-apoptotic MDM2 and Bcl-2 was significantly elevated while activity of pro-apoptotic Bax was decreased in RCC as compared with normal kidney tissues. Bax expression was positively correlated with patient age. Significant association has been detected between the evaluated markers and cancer clinical parameters like: tumor stage, grade, lymph node and distant metastases. The markers' activity was associates with the tumor morphological features, in particular: presence of tumor necrosis and microvascular invasion. Disease recurrence and 5-year patient survival were associated with the markers' activity. Cox regression analyses have shown that tumor size, pathological stage and grade are the risk factors for disease recurrence and patient death. Expression of MDM2 and Bcl-2 is significantly up-regulated, while Bax is down-regulated in RCC as compared with normal kidney tissue. Intensity of the markers'activities is associated with the tumor pathological and clinical parameters (stage, grade, lymph node and distant metastases, tumor recurrence and patient survival). Further studies with more patients and longer follow-up will uncover the clinical importance of the evaluated markers in RCC. PMID:26719546

  16. Identification of New Herpesvirus Gene Homologs in the Human Genome

    OpenAIRE

    Holzerlandt, Ria; Orengo, Christine; Kellam, Paul; Albà, M. Mar

    2002-01-01

    Viruses are intracellular parasites that use many cellular pathways during their replication. Large DNA viruses, such as herpesviruses, have captured a repertoire of cellular genes to block or mimic host immune responses, apoptosis regulation, and cell-cycle control mechanisms. We have conducted a systematic search for all homologs of herpesvirus proteins in the human genome using position-specific scoring matrices representing herpesvirus protein sequence domains, and pair-wise sequence comp...

  17. Loss of anti-Bax function in Gerstmann-Straussler-Scheinker syndrome-associated prion protein mutants.

    Directory of Open Access Journals (Sweden)

    Julie Jodoin

    Full Text Available Previously, we have shown the loss of anti-Bax function in Creutzfeldt Jakob disease (CJD-associated prion protein (PrP mutants that are unable to generate cytosolic PrP (CyPrP. To determine if the anti-Bax function of PrP modulates the manifestation of prion diseases, we further investigated the anti-Bax function of eight familial Gerstmann-Sträussler-Scheinker Syndrome (GSS-associated PrP mutants. These PrP mutants contained their respective methionine ((M or valine ((V at codon 129. All of the mutants lost their ability to prevent Bax-mediated chromatin condensation or DNA fragmentation in primary human neurons. In the breast carcinoma MCF-7 cells, the F198S(V, D202N(V, P102L(V and Q217R(V retained, whereas the P102L(M, P105L(V, Y145stop(M and Q212P(M PrP mutants lost their ability to inhibit Bax-mediated condensed chromatin. The inhibition of Bax-mediated condensed chromatin depended on the ability of the mutants to generate cytosolic PrP. However, except for the P102L(V, none of the mutants significantly inhibited Bax-mediated caspase activation. These results show that the cytosolic PrP generated from the GSS mutants is not as efficient as wild type PrP in inhibiting Bax-mediated cell death. Furthermore, these results indicate that the anti-Bax function is also disrupted in GSS-associated PrP mutants and is not associated with the difference between CJD and GSS.

  18. Study of Ganoderma lucidum spores on pentylenetetrazol activation of hippocampal neurons bax expression%灵芝孢子粉对戊四氮活化海马神经细胞bax表达的研究

    Institute of Scientific and Technical Information of China (English)

    张金波; 王淑秋; 张淑红; 金岳雷; 朱金玲; 刘爽

    2012-01-01

    目的 本实验研究灵芝孢子粉对戊四氮活化大鼠海马神经细胞bax表达变化的影响,进一步探讨灵芝孢子粉的作用机制和癫痫与海马神经细胞凋亡调控基因之间的关系.方法 通过制备癫痫模型和RT-PCR检测正常对照组、癫痫模型组和灵芝孢子粉用药组bax的表达.结果 癫痫模型组和灵芝孢子粉用药组bax的表达较正常对照组(0.17±0.04)均升高;其中癫痫模型组(0.66±0.09)bax的表达水平与对照组(0.17±0.04)比较明显升高(P<0.01),灵芝孢子粉用药组(0.47±0.1)bax的表达水平与癫痫模型组比较明显降低(P<0.01),差异有统计学意义.结论 本研究结果证实,在PTZ点燃癫痫后,模型组和治疗组促凋亡基因bax表达较正常对照组显著升高,表明bax基因在细胞凋亡的调控过程中起到促进作用,而给予灵芝孢子粉治疗后,bax的表达与模型组比较显著降低,提示灵芝孢子粉有效成份能充分作用于脑组织,可以调控bax的表达,借以发挥抗凋亡的神经保护作用.%Objective; This study is Ganoderma lucidum spores on pentylenetetrazol activation of rat hippocampal neurons bax expression changes to further explore the relationship between the action mechanism of the Ganoderma lucidum spores and epilepsy with apoptosis regulatory genes of hippocampal neural. Methods; Through the preparation of the epileptic model and RT — PCR to detect of bax expression in the normal control group, epilepsy model group and Ganoderma spore powder medication group. Results;/ Bax expression in epilepsy model group and Ganoderma lucidum spores medication group were higher than in the normal control group (0. 17 ± 0. 04) ; including bax expression level in epilepsy model group (0. 66 ± 0. 09) was significantly higher than in the control group (0. 17 ± 0.04) (P<0. 01) , bax expression level in Ganoderma spore powder medication group (0.47 ± 0. 1 ) were significantly lower than in epilepsy model group

  19. Comparative study of Bax, Bcl-2 protein expression in villi structure during normal pregnancy and missed abortion%正常早孕与稽留流产绒毛组织结构中Bax和Bcl-2蛋白表达的对比研究

    Institute of Scientific and Technical Information of China (English)

    杨兴爽

    2014-01-01

    Objective To study apoptosis regulating proteins Bcl-2 and Bax in normal pregnancy and missed abortion villi structure and its significance, and explore the reasons for missed abortion. Methods Choose our hospital patients missed abortion and early pregnancy abortion patients, 60 cases in total, divided into group A and group B. Immediately after abortion, embryonic villi specimens sent to pathology. Light microscopy were applied in each group villi morphological changes in the structure, while applying immunohistochemical methods and computer image analysis system to detect Bax, Bcl-2 expression in each group villi and cell apoptosis. Results Missed abortion group trophoblast cell apoptosis index was (33.32±0.79)%, was significantly higher in group B(18.90±0.63)%, difference was statistically significant (P<0.01). Bax and Bcl-2 in the two groups syncytiotrophoblast cells show positive rate. In missed abortion group, Bax positive rate increased, Bcl-2 positive rate of decline, Bcl-2/Bax ratio increased, differences were statistically significant (P<0.01). Conclusion The increasing positive expression rate of Bax and the decreasing positive expression rate of Bcl-2 in decidua villi can lead to villous syncytiotrophoblast cells increased significantly,and further lead to missed abortion.%目的:研究凋亡调控蛋白Bcl-2和Bax在正常早孕与稽留流产绒毛组织结构中的表达及其意义,探讨稽留流产原因。方法选择本院就诊的稽留流产患者和早孕人工流产患者各60例,分为A组、B组。流产后立即留取胚胎绒毛组织送病理。分别应用光镜观察各组绒毛组织细胞形态结构的改变;同时应用免疫组织化学方法和计算机图文分析系统检测Bax、Bcl-2在各组绒毛的表达及细胞凋亡情况。结果稽留流产组绒毛滋养细胞中凋亡指数为(33.32±0.79)%,明显高于对照组B组的(18.90±0.63)%,差异有统计学意义(P<0.01)。Bax和Bcl-2在两组合体滋养

  20. Bax translocation mediated mitochondrial apoptosis and caspase dependent photosensitizing effect of Ficus religiosa on cancer cells.

    Directory of Open Access Journals (Sweden)

    Jazir Haneef

    Full Text Available The main aim of the present work was to investigate the potential effect of acetone extract of Ficus religosa leaf (FAE in multiple apoptosis signalling in human breast cancer cells. FAE treatment significantly induced dose and time dependent, irreversible inhibition of breast cancer cell growth with moderate toxicity to normal breast epithelial cells. This observation was validated using Sulforhodamine B assay. Cell cycle analysis by Flow cytometry showed cell cycle arrest in G1 phase and induction of sub-G0 peak. FAE induced chromatin condensation and displayed an increase in apoptotic population in Annexin V-FITC/PI (Fluorescein isothiocyanate/Propidium iodide double staining. FAE stimulated the loss of mitochondrial membrane potential in multiple breast cancer cell lines when compared to normal diploid cells. To understand the role of Bax in FAE induced apoptosis, we employed a sensitive cell based platform of MCF-7 cells expressing Bax-EGFP. Bax translocation to mitochondria was accompanied by the disruption of mitochondrial membrane potential and marked elevation in LEHDase activity (Caspase 9. Consistent with this data, FAE induced Caspase activation as evidenced by ratio change in FRET Caspase sensor expressing MCF-7 cell line and cleavage of prominent Caspases and PARP. Interestingly, FAE accelerated cell death in a mitochondrial dependent manner in continuous live cell imaging mode indicating its possible photosensitizing effect. Intracellular generation of reactive oxygen species (ROS by FAE played a critical role in mediating apoptotic cell death and photosensitizing activity. FAE induced dose and time dependent inhibition of cancer cell growth which was associated with Bax translocation and mitochondria mediated apoptosis with the activation of Caspase 9 dependent Caspase cascade. FAE also possessed strong photosensitizing effect on cancer cell line that was mediated through rapid mitochondrial transmembrane potential loss and

  1. Bax translocation mediated mitochondrial apoptosis and caspase dependent photosensitizing effect of Ficus religiosa on cancer cells.

    Science.gov (United States)

    Haneef, Jazir; Parvathy, Muraleedharan; M, Parvathy; Thankayyan R, Santhosh Kumar; Sithul, Hima; Sreeharshan, Sreeja

    2012-01-01

    The main aim of the present work was to investigate the potential effect of acetone extract of Ficus religosa leaf (FAE) in multiple apoptosis signalling in human breast cancer cells. FAE treatment significantly induced dose and time dependent, irreversible inhibition of breast cancer cell growth with moderate toxicity to normal breast epithelial cells. This observation was validated using Sulforhodamine B assay. Cell cycle analysis by Flow cytometry showed cell cycle arrest in G1 phase and induction of sub-G0 peak. FAE induced chromatin condensation and displayed an increase in apoptotic population in Annexin V-FITC/PI (Fluorescein isothiocyanate/Propidium iodide) double staining. FAE stimulated the loss of mitochondrial membrane potential in multiple breast cancer cell lines when compared to normal diploid cells. To understand the role of Bax in FAE induced apoptosis, we employed a sensitive cell based platform of MCF-7 cells expressing Bax-EGFP. Bax translocation to mitochondria was accompanied by the disruption of mitochondrial membrane potential and marked elevation in LEHDase activity (Caspase 9). Consistent with this data, FAE induced Caspase activation as evidenced by ratio change in FRET Caspase sensor expressing MCF-7 cell line and cleavage of prominent Caspases and PARP. Interestingly, FAE accelerated cell death in a mitochondrial dependent manner in continuous live cell imaging mode indicating its possible photosensitizing effect. Intracellular generation of reactive oxygen species (ROS) by FAE played a critical role in mediating apoptotic cell death and photosensitizing activity. FAE induced dose and time dependent inhibition of cancer cell growth which was associated with Bax translocation and mitochondria mediated apoptosis with the activation of Caspase 9 dependent Caspase cascade. FAE also possessed strong photosensitizing effect on cancer cell line that was mediated through rapid mitochondrial transmembrane potential loss and partial Caspase

  2. Influence of neurotrophin-3 on Bcl-2 and Bax expressions in spinal cord injury of rats

    Institute of Scientific and Technical Information of China (English)

    GUO Shu-zhang; JIANG Tao; REN Xian-jun

    2007-01-01

    Objective:To study the protective mechanisms of neurotrophin-3 (NT-3) on the spinal cord injury.Methods:Totally 105 SD rats were randomly divided into 3 groups:control group,experimental group and sham operation group.Rats from the former 2 groups were inflicted to animal model of acute spinal cord injury according to Allen's (WD) by situating a thin plastic tube in the subarachnoid space below the injury level for perfusion.Rats in experimental group received 20μl NT-3 (200 ng) from the tube at 0,4,8,12,24 h and 3,7 d after injury,and those in control group got an equal volume of normal saline at the same time.The animals in sham operation group only received opening vertebral plate and tube was put in subarachnoid space.The rats were sacrificed at 4,8,12,24 h and 3,7,14 d post injury (n=5).The expression levels of Bcl-2 and Bax proteins in spinal cord of rats were detected by immunohistochemistry assay.Results:The level of Bax protein in control group significantly increased as compared with those in sham operation group, and the peak reached at 8 h after spinal cord injury.The Bcl-2 proteins were always weakly positive.The Bax proteins in NT-3 group significantly decreased but the Bcl-2 proteins obviously increased as compared with those in control group.Conclusion:NT-3 can protect spinal cord from injury in vivo.One of the mechanisms is that NT-3 can inhibit abnormal expression of Bax protein,and increase the expression of Bcl-2 protein,then inhibit apoptosis after spinal cord injury.

  3. Effects of apoptosis-related proteins caspase-3, Bax and Bcl-2 on cerebral ischemia rats

    OpenAIRE

    Liu, Guangyi; Tao WANG; WANG, TINGING; Song, Jinming; Zhou, Zhen

    2013-01-01

    Neuron apoptosis is known to mediate a change of ethology following cerebral ischemia-reperfusion injury in rats. Additionally, Bcl-2, Bax and caspase-3 proteins may exert a significant effect on neuron injury. The aim of this study was to investigate the role, mechanism of action and clinical significance of these proteins in neuron apoptosis and functional impairment following cerebral ischemia-reperfusion injury in rats. Sixty male healthy adult Wistar rats were randomly assigned into cont...

  4. Electrical conductivity and defect chemistry of BaxSr1 - xCoyFe1 - yO3 - dBaxSr1−xCoyFe1−yO3− perovskites

    NARCIS (Netherlands)

    Yáng, Z.; Harvey, A.S.; Infortuna, A.; Schoonman, J.; Gauckler, L.J.

    2010-01-01

    Bulk BaxSr1 - xCoyFe1 - yO3 - dBaxSr1−xCoyFe1−yO3− compositions (BSCF) were synthesized by the solid-state reaction method. The electrical conductivity of ceramic bars was measured using a dc four-probe method as a function of temperature in air up to 970 °C. All compositions showed thermally activa

  5. Asiaticoside: attenuation of neurotoxicity induced by MPTP in a rat model of Parkinsonism via maintaining redox balance and up-regulating the ratio of Bcl-2/Bax.

    Science.gov (United States)

    Xu, Chang-Liang; Wang, Qi-Zhi; Sun, Ling-Mei; Li, Xiu-Min; Deng, Ji-Min; Li, Lu-Fan; Zhang, Jin; Xu, Rong; Ma, Shi-Ping

    2012-01-01

    In this study, we investigated the neuroprotective effects of asiaticoside, a triterpenoid saponin isolated from the Chinese medicinal herb Centella asiatica, in the rats model of Parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Rats were first injected with MPTP. One day after surgery, asiaticoside was administered and the behavioral tests were assessed. On 14th day, the rats were sacrificed, substantia nigra (SN) and striatum were dissected, and then dopamine (DA) and its metabolites in striatum and malonyldialdehyde (MDA) contents, reduced glutathione (GSH) level and gene expression level in SN were estimated. Treatment with asiaticoside was found to protect dopaminergic neuron by antagonizing MPTP induced neurotoxicity and to improve locomotor dysfunction. Asiaticoside significantly attenuated the MPTP-induced reduction of dopamine in the striatum. The content of MDA was significantly decreased while the GSH level was significantly increased in asiaticoside-treated groups. In addition, asiaticoside increased the Bcl-2/Bax ratio. These results indicated that asiaticoside was effective in reversing MPTP induced Parkinsonism via its neuroprotective effects including antioxidant activity, maintaining the metabolic balance of DA, and increasing ratio of Bcl-2/Bax.

  6. AxBAxB… pulsed atomic layer deposition: Numerical growth model and experiments

    Science.gov (United States)

    Muneshwar, Triratna; Cadien, Ken

    2016-02-01

    Atomic layer deposition (ALD) is widely used for the fabrication of advanced semiconductor devices and related nanoscale structures. During ALD, large precursor doses (>1000 L per pulse) are often required to achieve surface saturation, of which only a small fraction is utilized in film growth while the rest is pumped from the system. Since the metal precursor constitutes a significant cost of ALD, strategies to enhance precursor utilization are essential for the scaling of ALD processes. In the precursor reaction step, precursor physisorption is restricted by steric hindrance (mA1) from ligands on the precursor molecules. On reaction, some of these ligands are removed as by-products resulting in chemisorbed species with reduced steric hindrance (mA1 → mA2, where mA2 1, x ∈ I) short-pulses rather than a single pulse. A numerical first-order surface reaction kinetics growth model is presented and applied to study the effect of AxBAxB… pulsed ALD on the growth per cycle (GPC). The model calculations predict higher GPC for AxBAxB… pulsing than with ABAB… deposition. In agreement with the model predictions, with AxBAxB… pulsed deposition, the GPC was found to increase by ˜46% for ZrN plasma enhanced ALD (PEALD), ˜49% for HfO2 PEALD, and ˜8% for thermal Al2O3 ALD with respect to conventional ABAB… pulsed growth.

  7. Plumbagin reduces chronic lymphocytic leukemia cell survival by downregulation of Bcl-2 but upregulation of the Bax protein level.

    Science.gov (United States)

    Fu, Chunling; Gong, Yanqing; Shi, Xuanxuan; Sun, Zengtian; Niu, Mingshan; Sang, Wei; Xu, Linyan; Zhu, Feng; Wang, Ying; Xu, Kailin

    2016-09-01

    Chronic lymphocytic leukemia (CLL) is the most common leukemia in Western countries, and mainly originates from an accumulation of abnormal B cells caused by the dysregulation of cell proliferation and apoptosis rates. The aberration of apoptosis-related genes in CLL cells results in defective apoptosis of CLL cells in response to traditional therapeutic medicine. Plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone), a natural compound from Plumbago zeylinica, has been shown to exhibit pro-apoptotic activities in tumor cells. In the present study, we report that plumbagin effectively inhibited CLL cell viability with a lower dose compared to fludarabine, and inhibited cell proliferation in a dose-dependent manner. In addition, plumbagin promoted accumulation of MEC-1 cells in the S phase, and blocked cell cycle transition of HG3 cells from G0/G1 to S phase. Molecularly, plumbagin markedly induced CLL cell apoptosis through reduction of Bcl-2, but through an increase in the Bax protein level. These results suggest that plumbagin may be considered as a potential anticancer agent for CLL therapy. PMID:27461100

  8. Expression and Significance of Bcl-2, Bax, Fas and Caspase-3 in Different Phases of Human Hemangioma

    Institute of Scientific and Technical Information of China (English)

    YANG Hong; DENG Chenguo; SHEN Shengguo; ZHANG Duanlian; YUYing

    2006-01-01

    The relationship between Bcl-2, Bax, Fas, caspase-3 and development of hemangioma and the molecular mechanism was investigated. By using immunohistochemical S-P method, proliferating cell nuclear antigen was detected. According to the classification of Mulliken in combination with PCNA expression, 27 cases were identified as proliferating hemangioma and 22 cases as involutive hemangioma. Five normal skin tissues around the tumor tissue served as controls. By using immunohistochemical technique, the expression of Bcl-2, Bax, Fax and Caspase-3 was detected. The cells expressing Bcl-2, Bax, Fax and cappase-3 were identified as hemangioma endothelia by immunohistochemical staining of Ⅷ factor. The average absorbance (A) and average positive area rate of Bcl-2, Bax, Fas and caspase-3 expression were measured by using HPIAS-2000 imaging analysis system. The results showed that the expression of Bcl-2 in the endothelia of proliferating hemangioma was significantly higher that in involutive degenerative hemangioma endothelia and vascular endothelia of normal skin tissue (P<0.01). The expression of Bax, Fas and Caspase-3 in the endothelia of involutive hemangioma was obviously higher than in the endothelia of proliferating hemangioma and normal skin tissue (P<0.01). The expression of BAx and Fas in endothelia of proliferating hemangioma was higher than in those of normal skin tissue (P<0.05). It was suggested that Bcl-2,Bax, Fas and caspase-3 might be involved in the development and involution of hemangioma. Bcl-2 could promote the growth of hemangioma by inhibiting apoptosis of endothelia. Bax, Fas and caspase-3 promote the switch of hemangioma from proliferation to involution by inducing the apoptosis of hemangioma endothelia.

  9. Bax-induced apoptosis shortens the life span of DNA repair defect Ku70-knockout mice by inducing emphysema.

    Science.gov (United States)

    Matsuyama, Shigemi; Palmer, James; Bates, Adam; Poventud-Fuentes, Izmarie; Wong, Kelvin; Ngo, Justine; Matsuyama, Mieko

    2016-06-01

    Cells with DNA damage undergo apoptosis or cellular senescence if the damage cannot be repaired. Recent studies highlight that cellular senescence plays a major role in aging. However, age-associated diseases, including emphysema and neurodegenerative disorders, are caused by apoptosis of lung alveolar epithelial cells and neurons, respectively. Therefore, enhanced apoptosis also promotes aging and shortens the life span depending on the cell type. Recently, we reported that ku70(-) (/) (-)bax(-) (/) (-) and ku70(-) (/) (-)bax(+/) (-) mice showed significantly extended life span in comparison with ku70(-) (/) (-)bax(+/+) mice. Ku70 is essential for non-homologous end joining pathway for DNA double strand break repair, and Bax plays an important role in apoptosis. Our study suggests that Bax-induced apoptosis has a significant impact on shortening the life span of ku70(-) (/) (-) mice, which are defective in one of DNA repair pathways. The lung alveolar space gradually enlarges during aging, both in mouse and human, and this age-dependent change results in the decrease of respiration capacity during aging that can lead to emphysema in more severe cases. We found that emphysema occurred in ku70(-) (/) (-) mice at the age of three-months old, and that Bax deficiency was able to suppress it. These results suggest that Bax-mediated apoptosis induces emphysema in ku70(-) (/) (-) mice. We also found that the number of cells, including bronchiolar epithelial cells and type 2 alveolar epithelial cells, shows a higher DNA double strand break damage response in ku70 KO mouse lung than in wild type. Recent studies suggest that non-homologous end joining activity decreases with increased age in mouse and rat model. Together, we hypothesize that the decline of Ku70-dependent DNA repair activity in lung alveolar epithelial cells is one of the causes of age-dependent decline of lung function resulting from excess Bax-mediated apoptosis of lung alveolar epithelial cells (and their

  10. Significance and expression of Bax, Survivin and p53 in gastric carcinoma and precancerous lesions using tissue microarray%利用组织芯片技术研究胃癌及其癌前病变中Bax、p53、Survivin表达的关系及意义

    Institute of Scientific and Technical Information of China (English)

    Yuping Xiao; Zhi Lin; Lili Mao; Dongying Wu; Yujia Gao; Hongwei Sun; Yan Xin

    2007-01-01

    Objective: To explore the relationship between expressions of apoptosis-related protein Bax, Survivin and p53 and the molecular mechanisms of carcinogenesis and progression of gastric carcinoma. Methods: Tissue microarray and immunohistochemistry were used in this study. Results: The positive rate of Bax protein in gastric cancer (17.7%, 17/96) was significantly lower than those in adjacent normal mucosa (51%), intestinal metaplasia (69.2%) and dysplasia (75%), P < 0.01.The positive rate of Survivin expression in gastric cancer (80.6%, 89/98) was significantly higher than that in adjacent normal mucosa (3.9%), P < 0.01. The positive rates of Survivin expression in tumors with different organ metastases (in lymph node metastasis 86.2%, liver 100% and ovarian 100%) were statistically higher than in tumors without metastasis (64.3%), P <0.05. Bax expression was correlated with Survivin but not with mp53 that was closely related to Survivin expression (P < 0.05)in gastric cancer. Conclusion: The abnormal expressions of Bax, Survivin and mp53 were correlated with the tumorigenesis and progression of gastric carcinoma. P53 and Survivin genes may share the similar mechanism in regulating cell apoptosis,and because of the mutation, p53 gene may lower its down-regulation to Survivin expression.

  11. Associations of MMP-2, BAX, and Bcl-2 mRNA and Protein Expressions with Development of Atrial Fibrillation.

    Science.gov (United States)

    Diao, Shu-Ling; Xu, Hui-Pu; Zhang, Bei; Ma, Bao-Xin; Liu, Xian-Liang

    2016-01-01

    BACKGROUND To examine changes of mRNA and protein expressions of MMP-2, Bcl-2, and BAX in atrial fibrillation (AF) patients, and investigate the correlations among these 3 biomarkers. MATERIAL AND METHODS Rheumatic heart disease patients (n=158) undergoing cardiac surgical procedures for mitral valve repair or replacement were included as the AF group (n=123), containing paroxysmal AF (n=42), persistent AF (n=36), and permanent AF (n=45). Rheumatic heart disease patients with sinus rhythm (SR) (n=35) were enrolled as the SR group (control group). Immunohistochemistry, Western blot, and real-time polymerase chain reaction (PCR) were applied to detect the protein and mRNA expression levels of MMP-2, Bcl-2, and BAX. Apoptosis was observed with light and electron microscopes and detected by TdT-mediated dUTP nick-end labeling (TUNEL). RESULTS Compared with the SR group, the left atrial diameters (LADs), protein and mRNA expression levels of MMP-2 and BAX, apoptotic index (AI), and Bcl-2/BAX ratio were evidently increased in the 3 AF groups, but protein and mRNA expression levels of Bcl-2 decreased in the AF groups (all P<0.05). Correlation analysis found that MMP-2 protein expression levels was positively correlated with BAX expression, but negatively correlated with Bcl-2 expression levels. CONCLUSIONS Our study results suggest that elevated MMP-2 expression and disturbance balance of Bcl-2/BAX expressions may be associated with the development and maintenance of AF. MMP-2 may be involved in the development of AF through promoting BAX expressions and inhibiting Bcl-2. PMID:27141955

  12. BAX and tumor suppressor TRP53 are important in regulating mutagenesis in spermatogenic cells in mice.

    Science.gov (United States)

    Xu, Guogang; Vogel, Kristine S; McMahan, C Alex; Herbert, Damon C; Walter, Christi A

    2010-12-01

    During the first wave of spermatogenesis, and in response to ionizing radiation, elevated mutant frequencies are reduced to a low level by unidentified mechanisms. Apoptosis is occurring in the same time frame that the mutant frequency declines. We examined the role of apoptosis in regulating mutant frequency during spermatogenesis. Apoptosis and mutant frequencies were determined in spermatogenic cells obtained from Bax-null or Trp53-null mice. The results showed that spermatogenic lineage apoptosis was markedly decreased in Bax-null mice and was accompanied by a significantly increased spontaneous mutant frequency in seminiferous tubule cells compared to that of wild-type mice. Apoptosis profiles in the seminiferous tubules for Trp53-null were similar to control mice. Spontaneous mutant frequencies in pachytene spermatocytes and in round spermatids from Trp53-null mice were not significantly different from those of wild-type mice. However, epididymal spermatozoa from Trp53-null mice displayed a greater spontaneous mutant frequency compared to that from wild-type mice. A greater proportion of spontaneous transversions and a greater proportion of insertions/deletions 15 days after ionizing radiation were observed in Trp53-null mice compared to wild-type mice. Base excision repair activity in mixed germ cell nuclear extracts prepared from Trp53-null mice was significantly lower than that for wild-type controls. These data indicate that BAX-mediated apoptosis plays a significant role in regulating spontaneous mutagenesis in seminiferous tubule cells obtained from neonatal mice, whereas tumor suppressor TRP53 plays a significant role in regulating spontaneous mutagenesis between postmeiotic round spermatid and epididymal spermatozoon stages of spermiogenesis. PMID:20739667

  13. Expression of Ki-67, Bcl-2 and Bax in the First Trimester Abortion Materials

    Directory of Open Access Journals (Sweden)

    Ender DÜZCAN

    2010-01-01

    Full Text Available Objective: The aim of this study was to investigate possible similar or different mechanisms in recurrent and spontaneous abortion by evaluating immunohistochemical correlation between proliferation marker Ki-67, and apoptosis markers Bcl-2 and Bax in the fetal trophoblasts and maternal deciduas from abortion material.Material and Method: Eighty samples of curettage materials from 65 abortion patients histopathologically diagnosed “decidua showing Arias-Stella reaction and chorionic villi” or only “decidua showing Arias-Stella reaction” were included in the study. Hematoxylin&Eosin stained sections from all cases were re-evaluated and further stained immunohistochemically using antibodies against Ki-67, Bcl-2 and Bax.Results: Proliferation rate evaluated by Ki-67 expression both in the cytotrophoblastic cells and decidua was found to be significantly lower in spontaneous and recurrent abortions compared to evacuation abortion. The extent of Bcl-2 expression in syncytiotrophoblastic cells covering villous stroma was also decreased in spontaneous abortion. There were no significant differences between spontaneous and recurrent abortions in terms of Bcl-2 expression in syncytiotrophoblasts and Ki-67 proliferation index in cytotrophoblastic cells or decidua. Bax staining showed minimal decidual expression in a few spontaneous and recurrent abortions.Conclusion: We concluded that proliferation rate was decreased in fetal villous cytotrophoblasts and maternal deciduas in spontaneous and recurrent abortions. We also proposed that loss of Bcl-2 expression in syncytiotrophoblasts may cause abortion in a subset of cases. However, the data from spontaneous and recurrent abortions did not not support the presence of different mechanisms in both groups.

  14. Proteomic Profiling of Differentially Expressed Proteins from Bax inhibitor-1 Knockout and Wild Type Mice

    OpenAIRE

    Li, Bo; John C Reed; Kim, Hyung-Ryong; Chae, Han-Jung

    2012-01-01

    Bax inhibitor-1 (BI-1) is an anti-apoptotic protein located in the endoplasmic reticulum (ER). The role of BI-1 has been studied in different physiopathological models including ischemia, diabetes, liver regeneration and cancer. However, fundamental knowledge about the effects of BI-1 deletion on the proteome is lacking. To further explore this protein, we compared the levels of different proteins in bi-1−/− and bi-1+/+ mouse tissues by two-dimensional electrophoresis (2-DE) and mass spectrom...

  15. Temporal Alterations in Cellular Bax:Bcl-2 Ratio following Traumatic Brain Injury in the Rat

    OpenAIRE

    Raghupathi, Ramesh; Strauss, Kenneth I.; Zhang, Chen; Krajewski, Stanislaw; Reed, John C.; McIntosh, Tracy K.

    2003-01-01

    Cell death/survival following CNS injury may be a result of alterations in the intracellular ratio of death and survival factors. Using immunohistochemistry, Western analysis and in situ hybridization, the expression of the anti-cell death protein, Bcl-2, and the pro-cell death protein, Bax, was evaluated following lateral fluid-percussion (FP) brain injury of moderate severity (2.3–2.6 atm) in adult male Sprague-Dawley rats. By 2 h post-injury, a marked reduction of cellular Bcl-2-immunoreac...

  16. Effect of Flunarizine on the Expression of Bax mRNA in Hippocampus of Amygdala Kindling Seizures Rat%氟桂利嗪对杏仁核点燃鼠海马Bax mRNA表达的影响

    Institute of Scientific and Technical Information of China (English)

    梁代义; 伍国锋; 董佑忠; 庞成

    2005-01-01

    目的:观察氟桂利嗪对杏仁核点燃鼠癎性发作及海马促凋亡基因Bax mRNA表达的影响.方法:建立杏仁核点燃模型,予不同剂量氟桂利嗪灌喂点燃鼠.原位杂交法检测鼠脑海马Bax mRNA表达,图像分析软件测量阳性细胞平均吸光度.结果:正常大鼠海马存在少量Bax mRNA表达阳性细胞,点燃鼠海马各区Bax mRNA表达阳性细胞数及平均吸光度增加,氟桂利嗪处理后平均吸光度下降与剂量有关.结论:氟桂利嗪具有抗癫癎效应和拮抗点燃鼠海马Bax mRNA表达的作用.

  17. PUMA promotes Bax translocation in FOXO3a-dependent pathway during STS-induced apoptosis

    Science.gov (United States)

    Zhang, Yingjie; Chen, Qun

    2009-08-01

    PUMA (p53 up-regulated modulator of apoptosis, also called Bbc3) was first identified as a BH3-only Bcl-2 family protein that is transcriptionally up-regulated by p53 and activated upon p53-dependent apoptotic stimuli, such as treatment with DNA-damaging drugs or UV irradiation. Recently studies have been shown that Puma is also up-regulated in response to certain p53-independent apoptotic stimuli, such as growth factor deprivation or treatment with glucocorticoids or STS (staurosporine). However, the molecular mechanisms of PUMA up-regulation and how PUMA functions in response to p53-independent apoptotic stimuli remain poorly understood. In this study, based on real-time single cell analysis, flow cytometry and western blotting technique, we investigated the function of PUMA in living human lung adenocarcinoma cells (ASTC-a-1) after STS treatment. Our results show that FOXO3a was activated by STS stimulation and then translocated from cytosol to nucleus. The expression of PUMA was up-regulated via a FOXO3a-dependent manner after STS treatment, while p53 had little function in this process. Moreover, cell apoptosis and Bax translocation induced by STS were not blocked by Pifithrin-α (p53 inhibitor), which suggested that p53 was not involved in this signaling pathway. Taken together, these results indicate that PUMA promoted Bax translocation in a FOXO3a-dependment pathway during STS-induced apoptosis, while p53 was dispensable in this process.

  18. HIV-1 Vpr-induced apoptosis is cell cycle dependent and requires Bax but not ANT.

    Directory of Open Access Journals (Sweden)

    Joshua L Andersen

    2006-12-01

    Full Text Available The HIV-1 accessory protein viral protein R (Vpr causes G2 arrest and apoptosis in infected cells. We previously identified the DNA damage-signaling protein ATR as the cellular factor that mediates Vpr-induced G2 arrest and apoptosis. Here, we examine the mechanism of induction of apoptosis by Vpr and how it relates to induction of G2 arrest. We find that entry into G2 is a requirement for Vpr to induce apoptosis. We investigated the role of the mitochondrial permeability transition pore by knockdown of its essential component, the adenine nucleotide translocator. We found that Vpr-induced apoptosis was unaffected by knockdown of ANT. Instead, apoptosis is triggered through a different mitochondrial pore protein, Bax. In support of the idea that checkpoint activation and apoptosis induction are functionally linked, we show that Bax activation by Vpr was ablated when ATR or GADD45alpha was knocked down. Certain mutants of Vpr, such as R77Q and I74A, identified in long-term nonprogressors, have been proposed to inefficiently induce apoptosis while activating the G2 checkpoint in a normal manner. We tested the in vitro phenotypes of these mutants and found that their abilities to induce apoptosis and G2 arrest are indistinguishable from those of HIV-1NL4-3 vpr, providing additional support to the idea that G2 arrest and apoptosis induction are mechanistically linked.

  19. Quantitative expression analysis and prognostic significance of the BCL2-associated X gene in nasopharyngeal carcinoma: a retrospective cohort study

    International Nuclear Information System (INIS)

    Nasopharyngeal carcinoma (NPC) is a highly metastatic epithelial malignancy showing high prevalence in Southeast Asia and North Africa. The BCL2-associated X (BAX) gene encodes the most important pro-apoptotic member of the BCL2 family. We have recently shown that BCL2 and BCL2L12, two other members of the same apoptosis-related family, possess significant prognostic value in NPC. The objective of the current study was to analyze BAX mRNA expression in nasopharyngeal biopsies of NPC patients, and to assess its prognostic potential in this disease. Total RNA was isolated from 88 malignant and 9 hyperplastic nasopharyngeal biopsies, resected from Tunisian patients. After cDNA synthesis by reverse transcription of polyadenylated RNA, BAX mRNA expression was analyzed using a highly sensitive quantitative real-time polymerase chain reaction (qRT-PCR) method. Lower BAX mRNA levels were detected in NPC biopsies than in hyperplastic nasopharyngeal samples. BAX mRNA expression status was associated with low tumor extent, negative regional lymph node status, and absence of distant metastases. Kaplan-Meier survival analysis demonstrated that patients with BAX mRNA-positive NPC have significantly longer disease-free survival (DFS) and overall survival (OS). In accordance with these findings, Cox regression analysis revealed that BAX mRNA expression can be considered as a favorable prognostic indicator of DFS and OS in NPC, independent of their gender, age, tumor histology, tumor extent, and nodal status. Furthermore, NPC patients without distant metastases are less likely to relapse when their primary tumor is BAX mRNA-positive, compared to metastasis-free patients with a BAX-negative nasopharyngeal malignancy. This is the first study examining the potential clinical utility of BAX as a prognostic tumor biomarker in NPC. We provide evidence that BAX mRNA expression can be considered as an independent favorable prognostic indicator of DFS and OS in NPC

  20. Fas-mediated apoptosis regulates the composition of peripheral alphabeta T cell repertoire by constitutively purging out double negative T cells.

    Directory of Open Access Journals (Sweden)

    Abdiaziz S Mohamood

    Full Text Available BACKGROUND: The Fas pathway is a major regulator of T cell homeostasis, however, the T cell population that is controlled by the Fas pathway in vivo is poorly defined. Although CD4 and CD8 single positive (SP T cells are the two major T cell subsets in the periphery of wild type mice, the repertoire of mice bearing loss-of-function mutation in either Fas (lpr mice or Fas ligand (gld mice is predominated by CD4(-CD8(- double negative alphabeta T cells that also express B220 and generally referred to as B220+DN T cells. Despite extensive analysis, the basis of B220+DN T cell lymphoproliferation remains poorly understood. In this study we re-examined the issue of why T cell lymphoproliferation caused by gld mutation is predominated by B220+DN T cells. METHODOLOGY AND PRINCIPAL FINDINGS: We combined the following approaches to study this question: Gene transcript profiling, BrdU labeling, and apoptosis assays. Our results show that B220+DN T cells are proliferating and dying at exceptionally high rates than SP T cells in the steady state. The high proliferation rate is restricted to B220+DN T cells found in the gut epithelium whereas the high apoptosis rate occurred both in the gut epithelium and periphery. However, only in the periphery, apoptosis of B220+DN T cell is Fas-dependent. When the Fas pathway is genetically impaired, apoptosis of peripheral B220+DN T cells was reduced to a baseline level similar to that of SP T cells. Under these conditions of normalized apoptosis, B220+DN T cells progressively accumulate in the periphery, eventually resulting in B220+DN T cell lymphoproliferation. CONCLUSIONS/SIGNIFICANCE: The Fas pathway plays a critical role in regulating the tissue distribution of DN T cells through targeting and elimination of DN T cells from the periphery in the steady state. The results provide new insight into pathogenesis of DN T cell lymphoproliferation.

  1. Exhaustive Training Increases Uncoupling Protein 2 Expression and Decreases Bcl-2/Bax Ratio in Rat Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    W. Y. Liu

    2013-01-01

    Full Text Available This work investigates the effects of oxidative stress due to exhaustive training on uncoupling protein 2 (UCP2 and Bcl-2/Bax in rat skeletal muscles. A total of 18 Sprague-Dawley female rats were randomly divided into three groups: the control group (CON, the trained control group (TC, and the exhaustive trained group (ET. Malondialdehyde (MDA, superoxide dismutase (SOD, xanthine oxidase (XOD, ATPase, UCP2, and Bcl-2/Bax ratio in red gastrocnemius muscles were measured. Exhaustive training induced ROS increase in red gastrocnemius muscles, which led to a decrease in the cell antiapoptotic ability (Bcl-2/Bax ratio. An increase in UCP2 expression can reduce ROS production and affect mitochondrial energy production. Thus, oxidative stress plays a significant role in overtraining.

  2. BAX and BAK1 are dispensable for ABT-737-induced dissociation of the BCL2-BECN1 complex and autophagy.

    Science.gov (United States)

    Pedro, Jose Manuel Bravo-San; Wei, Yongjie; Sica, Valentina; Maiuri, Maria Chiara; Zou, Zhongju; Kroemer, Guido; Levine, Beth

    2015-01-01

    Disruption of the complex of BECN1 with BCL2 or BCL2L1/BCL-XL is an essential switch that turns on cellular autophagy in response to environmental stress or treatment with BH3 peptidomimetics. Recently, it has been proposed that BCL2 and BCL2L1/BCL-XL may inhibit autophagy indirectly through a mechanism dependent on the proapoptotic BCL2 family members, BAX and BAK1. Here we report that the BH3 mimetic, ABT-737, induces autophagy in parallel with disruption of BCL2-BECN1 binding in 2 different apoptosis-deficient cell types lacking BAX and BAK1, namely in mouse embryonic fibroblasts cells and in human colon cancer HCT116 cells. We conclude that the BH3 mimetic ABT-737 induces autophagy through a BAX and BAK1-independent mechanism that likely involves disruption of BECN1 binding to antiapoptotic BCL2 family members.

  3. Effects of genistein on neuronal apoptosis, and expression of Bcl-2 and Bax proteins in the hippocampus of ovariectomized rats

    Institute of Scientific and Technical Information of China (English)

    Yun Peng; Bo Jiang; Huiling Wu; Ruchun Dai; Liming Tan

    2012-01-01

    Genistein is one of several isoflavones that has a structure similar to 17β-estradiol, has a strong antioxidant effect, and a high affinity to estrogen receptors. At 15 weeks after ovariectomy, the expression of Bcl-2 in the hippocampus of rats decreased and Bax expression increased, with an obvious upregulation of apoptosis. However, intraperitoneal injection of genistein or 17β-estradiol for 15 consecutive weeks from the second day after operation upregulated Bcl-2 protein expression, downregulated Bax protein expression, and attenuated hippocampal neuron apoptosis. Our experimental findings indicate that long-term intervention with genistein can lead to a decrease in apoptosis in hippocampal neurons following ovariectomy, upregulate the expression of Bcl-2, and downregulate the expression of Bax. In addition, genistein and 17β-estradiol play equal anti-apoptotic and neuroprotective roles.

  4. Recombinant factor IX (BAX326) in previously treated paediatric patients with haemophilia B: a prospective clinical trial.

    Science.gov (United States)

    Urasinski, T; Stasyshyn, O; Andreeva, T; Rusen, L; Perina, F G; Oh, M S; Chapman, M; Pavlova, B G; Valenta-Singer, B; Abbuehl, B E

    2015-03-01

    A newly developed recombinant factor IX (BAX326(1) ) was investigated for prophylactic use in paediatric patients aged 96% of bleeds (100% of minor, 88.9% of moderate and 100% of major bleeds); the majority (88.5%) resolved after 1-2 infusions. Longer T1/2 and lower IR were observed in younger children (<6 years) compared to those aged 6 to 12 years. BAX326 administered as prophylactic treatment as well as for controlling bleeds is efficacious and safe in paediatric patients aged <12 years with haemophilia B. PMID:25495591

  5. Attenuation of p53 expression and Bax down-regulation during phorbol ester mediated inhibition of apoptosis

    OpenAIRE

    Meßmer, Udo K; Brüne, Bernhard

    1997-01-01

    Nitric oxide (NO) caused apoptotic cell death in murine RAW 264.7 macrophages. Associated with apoptotic morphology we observed p53 up-regulation and increased Bax expression. 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C (PKC) activator potently blocked NO-induced apoptosis. To gain insights into the mechanisms involved we investigated the effect of TPA on apoptotic conveying proteins such as p53 and Bax.TPA (100 nM) attentuated p53 up-regulation elicited by the NO-releasing...

  6. Comparison of automated BAX polymerase chain reaction and standard culture methods for detection of Listeria monocyogenes in blue crab meat (Callinectus sapidus) and blue crab processing plants

    Science.gov (United States)

    This study compared the BAX Polymerase Chain Reaction method (BAX PCR) with the Standard Culture Method (SCM) for detection of L. monocytogenes in blue crab meat and crab processing plants. The aim of this study was to address this data gap. Raw crabs, finished products and environmental sponge samp...

  7. Laquinimod decreases Bax expression and reduces caspase-6 activation in neurons.

    Science.gov (United States)

    Ehrnhoefer, Dagmar E; Caron, Nicholas S; Deng, Yu; Qiu, Xiaofan; Tsang, Michelle; Hayden, Michael R

    2016-09-01

    Laquinimod is an immunomodulatory compound that has shown neuroprotective benefits in clinical trials for multiple sclerosis. Laquinimod ameliorates both white and gray matter damage in human patients, and prevents axonal degeneration in animal models of multiple sclerosis. Axonal damage and white matter loss are a common feature shared between different neurodegenerative diseases. Caspase-6 activation plays an important role in axonal degeneration on the molecular level. Increased activity of caspase-6 has been demonstrated in brain tissue from presymptomatic Huntington disease mutation carriers, and it is an early marker of axonal dysfunction. Since laquinimod is currently undergoing a clinical trial in Huntington disease (LEGATO-HD, clinicaltrials.gov ID: NCT02215616), we set out to evaluate its impact on neuronal caspase-6 activation. We find that laquinimod ameliorates DNA-damage induced activation of caspase-6 in primary neuronal cultures. This is an indirect effect that is not mediated by direct inhibition of the enzyme. The investigation of potential caspase-6 activating mechanisms revealed that laquinimod reduces the expression of Bax, a pro-apoptotic molecule that causes mitochondrial cytochrome c release and caspase activation. Bax expression is furthermore increased in striatal tissues from the YAC128 mouse model of HD in an age-dependent manner. Our results demonstrate that laquinimod can directly downregulate neuronal apoptosis pathways relevant for axonal degeneration in addition to its known effects on astrocytes and microglia in the CNS. It targets a pathway that is relevant for the pathogenesis of HD, supporting the hypothesis that laquinimod may provide clinical benefit. PMID:27296315

  8. Relationship between expression of Bax and Bcl-2 proteins and apoptosis in radiation compound wound healing of rats

    Institute of Scientific and Technical Information of China (English)

    崔玉芳; 夏国伟; 付小兵; 杨红; 彭瑞云; 张莹; 谷庆阳; 高亚兵; 崔雪梅; 胡文华

    2003-01-01

    Objective: To study the relationship between the expression of Bax, Bcl-2 proteins, and apoptosis in radiation compound wound healing of rats.Methods: Apoptosis, Bax and Bcl-2 proteins were estimated by in situ terminal labeling (TUNEL) and immunohistochemical methods. Results: (1) Changes of the apoptosis in wound healing showed three typical characteristics: early occurrence, high frequency and delayed disappearance after radiation to rats when compared with those of simple wound group, which might be an important reason for radiation-induced delayed wound healing. (2) The expression of Bax protein increased evidently with the increment of apoptosis and showed a good corresponding relationship with the apoptotic frequency in the process of wound healing. While the expression of Bcl-2 protein decreased obviously as the apoptosis reached a maximum and showed increasing tendency up to normal level when the apoptosis decreased distinctively. Conclusions: Bax and Bcl-2 proteins play an important role in the apoptotic regulation of radiation compound wound healing in rats.

  9. Apoptosis and Bax expression are increased by coal dust in the polycyclic aromatic hydrocarbon-exposed lung

    Energy Technology Data Exchange (ETDEWEB)

    Ghanem, M.M.; Battelli, L.A.; Mercer, R.R.; Scabilloni, J.F.; Kashon, M.L.; Ma, J.Y.C.; Nath, J.; Hubbs, A.F.

    2006-09-15

    Miners inhaling respirable coal dust (CD) frequently develop coal workers' pneumoconiosis. Many coal miners are also exposed to polycyclic aromatic hydrocarbon (PAH) components of diesel engine exhaust and cigarette smoke, which may contribute to lung disease in these workers. Recently, apoptosis was reported to play a critical role in the development of another pneumoconiosis of miners, silicosis. In addition, CID was reported to suppress cytochrome P450 1A1 (CYP1A1) induction by PAHs. We exposed rats intratracheally to 0.0, 2.5, 10.0, 20.0, or 40.0 mg/rat CD and, 11 days later, to intraperitoneal P-naphthoflavone (BNF), a PAH. In another group of rats exposed to CD and BNF, caspase activity was inhibited by injection of the pan-caspase inhibitor Q-VD-OPH (quinoline-Val-Asp (OMe)-CH{sub 2}-OPH). In rats exposed to BNF, CD exposure increased alveolar expression of the proapoptotic mediator Bax but decreased CYP1A1 induction relative to BNF exposure alone. Pan-caspase inhibition decreased CD-associated Bax expression and apoptosis but did not restore CYP1A1 activity. Further, CD-induced lung inflammation and alveolar epithelial cell hypertrophy and hyperplasia were not suppressed by caspase inhibition. It is concluded that combined BNF and CD exposure increased Bax expression and apoptosis in the lung, but Bax and apoptosis were not the major determinants of early lung injury in this model.

  10. Immunohistochemical study of epidermal and dermal expression of Bcl-X, Bcl-2 and bax in psoriasis.

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Objective: To investigate the regulation of cell proliferation and apoptosis in psoriasis. Methods: The expressions of Bcl-X, Bcl-2 and Bax were studied with immunohistochemical technique (SP) in the lesional and non-lesional psoriatic skin. Results: There were significant overexpressions of Bcl-X in all layers of epidermis, inflammatory cells and vascular endothelia in dermis;

  11. Resveratrol induces apoptosis in human esophageal carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    Hai-Bo Zhou; Yun Yan; Ya-Ni Sun; Ju-Ren Zhu

    2003-01-01

    AIM: To investigate the apoptosis in esophageal cancer cells induced by resveratrol, and the relation between this apoptosis and expression of Bcl-2 and Bax.METHODS: In in vitro experiments, MTr assay was used to determine the cell growth inhibitory rate. Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of esophageal cancer cell line EC-9706 before and after the resveratrol treatment. Immunohistochemical staining was used to detect the expression of apoptosis-regulated gene Bcl-2 and Bax.RESULTS: Resveratrol inhibited the growth of esophageal cancer cell line EC-9706 in a dose-and time-dependent manner. Resveratrol induced EC-9706 cells to undergo apoptosis with typically apoptotic characteristics, including morphological changes of chromatin condensation, chromatin crescent formation, nucleus fragmentation and apoptotic body formation. TUNEL assay showed that after the for 24 to 96 hours, the AIs were apparently increased with treated time (P<0.05). Immunohistochemical staining showed that after the treatment of EC-9706 cells with proteins were apparently reduced with treated time (P<0.05)and the PRs of Bax proteins were apparently increased with treated time (P<0.05).CONCLUSION: Resveratrol is able to induce the apoptosisin esophageal cancer. This apoptosis may be mediated by down-regulating the apoptosis-regulated gene Bcl-2 and upregulating the expression of apoptosis-regulated gene bax.

  12. Expression of P53, P21/WAF/CIP, BCL-2, BAX, BCL-X, and BAK in radiation-induced apoptosis in testicular germ cell tumor lines

    International Nuclear Information System (INIS)

    Purpose: Testicular germ cell tumors (TGCTs) represent one of the few tumor types that are curable by antineoplastic therapy, probably due to the high sensitivity of this neoplasm to induction of apoptosis by chemotherapeutic agents and/or ionizing radiation. Here, we tested cell susceptibility to radiation-induced apoptosis in a panel of TGCT cell lines and attempted to correlate this with the known potentially relevant molecular determinants (p53 gene status and Bcl-2 family proteins) of apoptosis. Methods and Materials: Induction of apoptosis by γ-radiation was morphologically recognized in NT2, NCCIT, S2, and 2102 EP using Hoechst/PI staining and additionally confirmed by Western blot analysis of PARP cleavage. The p53 gene status was estimated by sequence analysis. Expression of p21/WAF/CIP was determined by Northern blot analysis and immunoblotting was used to monitor p53, Bax, Bcl-2, Bcl-x, and Bak protein levels. In vitro colony formation was studied to establish clonogenic survival curves. Results: NT2 and NCCIT appeared to be susceptible for radiation-induced apoptosis, contrasting 2102 EP and S2 which were highly resistant. Sequence analysis showed that NT2, S2, and 2102 EP are homozygous for wild-type p53 (wtp53), whereas NCCIT contains mutant p53 (mtp53). NT2 and 2102 EP cells showed radiation-induced p53 upregulation, while NCCIT (mtp53) and S2 (no p53 protein) cells did not. Consistently, γ-radiation-induced DNA damage resulted in a p53-dependent transactivation of the p21/WAF/CIP gene in NT2 and 2102 EP, but not in mtp53-containing NCCIT cells and p53 nonexpressing S2 cells. Constitutive expression of Bax, Bcl-2, Bcl-x, and Bak was not affected by radiation and showed no correlation with cell susceptibility to radiation-induced apoptosis. A discrepancy was found between apoptosis and reproductive death. Conclusions: The present study revealed that: i) the presence of wtp53 may not be absolutely required for the hypersensitivity for radiation

  13. Experession of Bax in Lung Cancer Cell Apoptosis Induced by Peroxisome Proliferator-activated Receptor-γ Anoists

    Institute of Scientific and Technical Information of China (English)

    ZAHNGMin; BAIMing; 等

    2002-01-01

    Objective:To discuss the relationship between Bax expression level and lung cancer cell apoptosis induced by peroxisome proliferator-activated receptor-γ(PPAR-γ) agonists.Methods:RT-PCR and Western blot analyis were used to detect PPAR-γ expression in the lung cancer cells,and TUNEL was used to detect apoptosis induced by PPAR-γ agnoists,while in situ hybridization and immunohistochemistry were used to monitor the changes of Bax mRNA and protein expression levels after apoptosis induced.Results PPAR-γ expression was detectable in two kinds lung cancer cells (including Non-small cell lung cancer and small cell lung cancer) ,and PPAR-γ agonists could inhibit lung cancer growth through inducing apoptosis.The apoptostic rates in control group,15d-PGJ2 group and cilitazone group were (1.86±0.49)%,(25.8±2.9)±,and (17.3±1.9)%,(P<0.01)respectively;Bax mRNA expression rates in the three groups were (8.75± 1.36)%,(66.2±12.86)%,and(29.5±6.5)%(P<0.01)respectively;Bax protein expression rates in the three groups were(9.2±1.45)%,(63±10.4)%,and (34.5±6.0)%(P<0.05) respectively.Conclusion PPAR-y is predicted to be a new targes in treating lung cancer in the future,and Bax is most likely to work in treating lung cancer apoptosis induced by PPAR-y agnoists as a factor to induce apoptosis.

  14. Gene

    Data.gov (United States)

    U.S. Department of Health & Human Services — Gene integrates information from a wide range of species. A record may include nomenclature, Reference Sequences (RefSeqs), maps, pathways, variations, phenotypes,...

  15. Boron neutron capture therapy induces apoptosis of glioma cells through Bcl-2/Bax

    Directory of Open Access Journals (Sweden)

    Mao Xinggang

    2010-12-01

    Full Text Available Abstract Background Boron neutron capture therapy (BNCT is an alternative treatment modality for patients with glioma. The aim of this study was to determine whether induction of apoptosis contributes to the main therapeutic efficacy of BNCT and to compare the relative biological effect (RBE of BNCT, γ-ray and reactor neutron irradiation. Methods The neutron beam was obtained from the Xi'an Pulsed Reactor (XAPR and γ-rays were obtained from [60Co] γ source of the Fourth Military Medical University (FMMU in China. Human glioma cells (the U87, U251, and SHG44 cell lines were irradiated by neutron beams at the XAPR or [60Co] γ-rays at the FMMU with different protocols: Group A included control nonirradiated cells; Group B included cells treated with 4 Gy of [60Co] γ-rays; Group C included cells treated with 8 Gy of [60Co] γ-rays; Group D included cells treated with 4 Gy BPA (p-borono-phenylalanine-BNCT; Group E included cells treated with 8 Gy BPA-BNCT; Group F included cells irradiated in the reactor for the same treatment period as used for Group D; Group G included cells irradiated in the reactor for the same treatment period as used for Group E; Group H included cells irradiated with 4 Gy in the reactor; and Group I included cells irradiated with 8 Gy in the reactor. Cell survival was determined using the 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium (MTT cytotoxicity assay. The morphology of cells was detected by Hoechst33342 staining and transmission electron microscope (TEM. The apoptosis rate was detected by flow cytometer (FCM. The level of Bcl-2 and Bax protein was measured by western blot analysis. Results Proliferation of U87, U251, and SHG44 cells was much more strongly inhibited by BPA-BNCT than by irradiation with [60Co] γ-rays (P 60Co] γ-rays (P P Conclusions Compared with ��-ray and reactor neutron irradiation, a higher RBE can be achieved upon treatment of glioma cells with BNCT. Glioma cell apoptosis induced by

  16. Effect of Achyranthes bidentata polysaccharides on the expression of BCL-2 and bax in hepatic tissues after exhaustive exercise in rats.

    Science.gov (United States)

    Lin, Jinyang; Zhang, Zhuoying; Shan, Ying

    2010-01-01

    This study aims to assess the effects of Achyranthes bidentata polysaccharides (ABPS) on the expression of bcl-2 and bax in hepatic tissues after exhaustive exercise in order to provide theoretical support for the application of ABPS in the field of sports nutrition. Thirty male Sprague-Dawley rats were randomized into three groups, each consisting of 10 rats: Normal control group (NCG), Exhausting exercises control group (EECG), ABPS treated group (ATG). ABPS were fed orally by gastric intubation to rats of ABPS treated group (ATG) once daily for 7 days. Control animals (EECG and NCG) received the same amount of isotonic sodium chloride solution. Exhaustive exercise was performed on a rodent treadmill. The SP (streptavidin peroxidase) method for immunohistochemical staining was adopted to test the protein expression of bax and bcl-2 in the hepatic tissues of the rats. Exhausting exercises increased bax protein expression of hepatic tissues of rats and bax/bcl-2 ratio dramatically, but a decreased bcl-2 protein expression. In the rats fed ABPS orally by gastric intubation, the bax protein expression and bax/bcl-2 ratio obviously decreased, while bcl-2 protein expression increased. The result indicated that bax and bcl-2 co-regulated the exercise-induced hepatocyte apoptosis. Feeding ABPS orally by gastric intubation to rats can inhibit the hepatocyte apoptosis in exhaustive exercise. PMID:21731162

  17. BIM (BCL-2 interacting mediator of cell death) SAHB (stabilized α helix of BCL2) not always convinces BAX (BCL-2-associated X protein) for apoptosis.

    Science.gov (United States)

    Verma, Sharad; Goyal, Sukriti; Tyagi, Chetna; Jamal, Salma; Singh, Aditi; Grover, Abhinav

    2016-06-01

    The interaction of BAX (BCL-2-associated X protein) with BIM (BCL-2 interacting mediator of cell death) SAHB (stabilized α helix of BCL2) directly initiates BAX-mediated mitochondrial apoptosis. This molecular dynamics study reveals that BIM SAHB forms a stable complex with BAX but it remains in a non-functional conformation. N terminal of BAX folds towards the core which has been reported exposed in the functional monomer. The α1-α2 loop, which has been reported in open conformation in functional BAX, acquires a closed conformation during the simulation. BH3/α2 remains less exposed as compared to initial structure. The hydrophobic residues of BIM accommodates in the rear pocket of BAX during the simulation. A steep decrease in radius of gyration and solvent accessible surface area (SASA) indicates the complex folding to acquire a more stable but inactive conformation. Further the covariance matrix reveals that the backbone atoms' motions favour the inactive conformation of the complex. This is the first report on the non-functional BAX-BIM SAHB complex by molecular dynamics simulation in the best of our knowledge. PMID:27262527

  18. Titanium dioxide induces apoptotic cell death through reactive oxygen species-mediated Fas upregulation and Bax activation

    Directory of Open Access Journals (Sweden)

    Yoon TH

    2012-03-01

    Full Text Available Ki-Chun Yoo1, Chang-Hwan Yoon1, Dongwook Kwon2, Kyung-Hwan Hyun1, Soo Jung Woo1, Rae-Kwon Kim1, Eun-Jung Lim1, Yongjoon Suh1, Min-Jung Kim1, Tae Hyun Yoon2, Su-Jae Lee11Laboratory of Molecular Biochemistry, 2Laboratory of Nanoscale Characterization and Environmental Chemistry, Department of Chemistry, Hanyang University, Seoul, Republic of KoreaBackground: Titanium dioxide (TiO2 has been widely used in many areas, including biomedicine, cosmetics, and environmental engineering. Recently, it has become evident that some TiO2 particles have a considerable cytotoxic effect in normal human cells. However, the molecular basis for the cytotoxicity of TiO2 has yet to be defined.Methods and results: In this study, we demonstrated that combined treatment with TiO2 nanoparticles sized less than 100 nm and ultraviolet A irradiation induces apoptotic cell death through reactive oxygen species-dependent upregulation of Fas and conformational activation of Bax in normal human cells. Treatment with P25 TiO2 nanoparticles with a hydrodynamic size distribution centered around 70 nm (TiO2P25–70 together with ultraviolet A irradiation-induced caspase-dependent apoptotic cell death, accompanied by transcriptional upregulation of the death receptor, Fas, and conformational activation of Bax. In line with these results, knockdown of either Fas or Bax with specific siRNA significantly inhibited TiO2-induced apoptotic cell death. Moreover, inhibition of reactive oxygen species with an antioxidant, N-acetyl-L-cysteine, clearly suppressed upregulation of Fas, conformational activation of Bax, and subsequent apoptotic cell death in response to combination treatment using TiO2P25–70 and ultraviolet A irradiation.Conclusion: These results indicate that sub-100 nm sized TiO2 treatment under ultraviolet A irradiation induces apoptotic cell death through reactive oxygen species-mediated upregulation of the death receptor, Fas, and activation of the preapoptotic protein

  19. Effects of erythropoietin on the expression of tumor necrosis factor-alpha and Bax after facial nerve axotomy in rats

    Institute of Scientific and Technical Information of China (English)

    Wei Zhang; Shengyu Lü; Ziying Yu; Ming Bi; Bin Sun

    2011-01-01

    This study sought to evaluate the effect of high-dose erythropoietin (EPO; 5 000 IU/kg) on the expression of tumor necrosis factor-alpha (TNF-α) and Bax in the facial nucleus after facial nerve transection in rats. A total of 42 Wistar rats of both genders were used in this study, and 40 rats were randomly divided into 2 groups: EPO group and model group. The EPO group was treated with EPO once a day for 5 days at a dose of 5 000 IU/kg body weight. The model group was treated with saline of the same amount. At day 3 after EPO (or saline) treatment, the right facial nerves of the 40 rats were transected at the level of the stylomastoid foramen, with the left sides untreated. The remaining 2 rats that did not undergo axotomy served as the control group. The surviving motor neurons in operated rats were counted in coronal paraffin sections of the facial nucleus. The expression of TNF-α and Bax in the facial nucleus was detected by immunohistochemical staining at days 3, 7, 14, 21, and 28 after axotomy. At days 14, 21, and 28 after facial nerve axotomy, a significantly greater proportion of facial motor neurons survived in the EPO group than in the model group. After axotomy, the expression of TNF-α and Bax increased in motor neurons in both the EPO and the model groups. TNF-α expression reached its peak level at day 14 after axotomy, while Bax expression reached its peak level at day 21. TNF-α expression was much lower in the EPO group than in the model group at all time points. No significant difference in Bax expression was found between the EPO and the model groups. These results indicate that high-dose EPO treatment attenuates the increase in TNF-α expression in the facial nucleus and reduces the loss of motor neurons after facial nerve transection in rats. However, high-dose EPO treatment has little effect on Bax expression.

  20. Direct proof of static charge stripe correlations in La2-xBaxCuO4

    Science.gov (United States)

    Chen, X. M.; Thampy, V.; Mazzoli, C.; Barbour, A.; Gu, G.; Hill, J. P.; Tranquada, J. M.; Dean, M. P. M.; Wilkins, S. B.

    The nature of charge stripe order in the cuprates, and in particular whether the stripes are static or dynamic, is a key issue in understanding the relationship between stripes and superconductivity. In La2-xBaxCuO4 (LBCO) a low temperature structural distortion is widely believed to pin stripes into fixed, static domains, but such an assertion has never been directly verified. We performed resonant soft x-ray photon correlation spectroscopy (XPCS) to probe the charge order Bragg peak of 1/8 doped LBCO. At low temperatures, we observe time-independent x-ray speckle patterns persisting for more than three hours, proving the static nature of the stripes and we go on to discuss how stripe order melts with increasing temperature. Our results demonstrate that the combination of XPCS with diffraction limited light sources such as the National Synchrotron Light Source II can probe the dynamics of even subtle order parameters such as stripes in the cuprates. Work performed at Brookhaven National Laboratory was supported by the US Department of Energy, Division of Materials Science, under Contract No. DE-AC02-98CH10886. Use of the National Synchrotron Light Source II was supported under Contract No. DE-SC0012704.

  1. Infrared Spectra of La0.65BaxMnO3-δ Oxides

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The infrared spectra of La0.65BaxMnO3-d (x = 0.35, 0.33 and 0.30) were investigated experimentally. The result shows that the sample La0.65Ba0.33MnO3-d has the largest Curie temperature and the smallest resistivity and wave number of the stretching vibration mode of MnO6 octahedron at 300 K among the investigated samples. However, the absorption strength for the stretching vibration mode of MnO6 octahedron in La0.65Ba0.35MnO3-d is stronger for paramagnetic phase than that for ferromagnetic phase, which may be connected with the reducing of the dynamic incoherent Jahn-Teller distortion below Curie temperature. In addition, the large shift of wave number for the stretching mode at the temperatures from 293 to 423 K has been observed in La0.65Ba0.35MnO3-d, which may be due to the increase of the Mn-O bond length with temperature increasing.

  2. Human ribosomal protein L9 is a Bax suppressor that promotes cell survival in yeast.

    Science.gov (United States)

    Eid, Rawan; Sheibani, Sara; Gharib, Nada; Lapointe, Jason F; Horowitz, Avital; Vali, Hojatollah; Mandato, Craig A; Greenwood, Michael T

    2014-05-01

    The identification of a human ribosomal protein L9 (hRPL9) cDNA as a sequence capable of suppressing the lethal effects of heterologously expressed murine Bax in yeast led us to investigate its antiapoptotic potential. Using growth and viability assays, we show that yeast cells heterologously expressing hRPL9 are resistant to the growth inhibitory and lethal effects of exogenously supplied copper, indicating that it has pro-survival properties. To explore potential mechanisms, we used yeast mutants defective in all three types of programmed cell death (apoptosis, necrosis, and autophagy). The ability to retain pro-survival function in all the mutants suggests that hRPL9 may regulate a common pro-death process. In contrast, the yeast RPL9 orthologues, RPL9A and RPL9B, have opposite effects when overexpressed in yeast. In effect, instead of showing resistance to stress, RPL9A and RPL9B overexpressing cells show reduced cell growth. Further analysis indicates that the effects of overexpressed RPL9A and RPL9B are not in themselves lethal, instead, they serve to increase cell doubling time. Thus, yeast RPL9s are more representative of RPs whose extra-ribosomal function is similar to that of tumor suppressors. Taken together, our results demonstrate that RPL9 represents a species- and sequence-specific regulator of cell growth and survival. PMID:24305165

  3. Prognostic Significance of Apoptosis Related Gene Family bcl-2 in Human Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    To study the prognostic effect of bcl-2 oncogene and its gene family members bax, bcl-x expression in breast cancer patients. Methods: expression of bcl-2, bax proteins in 91 human breast cancer tissue sections were studied by immunohistochemical method. Bcl-x1 mRNA expression in frozen tissues from 16 breast cancer patients were detected using Northern blot method. Results: bcl-2 protein positivity was found in 60/91 (65.9%) patients, and bax positivity 59/91 (64.8%). Bcl-2 and bax expression levels were associated with apoptotic index(AI), histological grade, axillary lymph node metastasis, postoperative local recurrence and metastasis. Bcl-2 expression was related to ER positivity. In univariate analysis for disease free survival (DFS), bcl-2 and bax protein levels, and Al were all found to have prognostic value. The result of Cox's model multivariate analysis showed that bcl-2 protein level was an independent prognostic factor. In 16 frozen breast cancer tissues, 8/16(50%) had higher level of bcl-x1 mRNA, which showed correlation with bcl-2 protein expression and axillary lymph node metastasis. Conclusion: The findings indicate that dysregulated expressions of bcl-2, bax and bcl-x1 apoptosis-related genes, suggestive of serious deregulation of apoptotic process, may contribute to the biologic aggressiveness of breast cancer. Bcl-2 protein is an independent indicator of prognosis in breast cancer patients.

  4. Interaction between Hsp60 and Bax in normal human myocardium and in myocardium affected by dilated cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Tykhonkova I. O.

    2009-04-01

    Full Text Available The main functional compartments of molecular chaperone Hsp60 are mitochondria and cytoplasm. Up to 30 % of Hsp60 are located in cytoplasm of cardiomyocytes. The interaction between molecular chaperone Hsp60 and proapoptotic Bax protein in the cytoplasmic fraction from normal human heart tissue has been revealed by co-immunoprecipitation in contrast to myocardium affected by dilated cardiomyopathy, where this interaction has not been observed

  5. BAX and Tumor Suppressor TRP53 Are Important in Regulating Mutagenesis in Spermatogenic Cells in Mice1

    OpenAIRE

    Xu, Guogang; Vogel, Kristine S.; McMahan, C. Alex; Herbert, Damon C.; Walter, Christi A.

    2010-01-01

    During the first wave of spermatogenesis, and in response to ionizing radiation, elevated mutant frequencies are reduced to a low level by unidentified mechanisms. Apoptosis is occurring in the same time frame that the mutant frequency declines. We examined the role of apoptosis in regulating mutant frequency during spermatogenesis. Apoptosis and mutant frequencies were determined in spermatogenic cells obtained from Bax-null or Trp53-null mice. The results showed that spermatogenic lineage a...

  6. The effect of octreotide and bromocriptine on expression of a pro-apoptotic Bax protein in rat prolactinoma.

    Directory of Open Access Journals (Sweden)

    Jolanta Kunert-Radek

    2004-03-01

    Full Text Available It is well established that disruption of apoptosis may lead to tumor initiation, progression or metastasis. It is also well documented that many anticancer drugs induce apoptosis. In the earlier studies, the dopamine D2 receptor agonist bromocriptine (BC and somatostatin analog octreotide (OCT were found to inhibit the growth of the estrogen-induced rat prolactinoma. Our previous investigations, applying the TUNEL method showed the involvement of the pro-apoptotic effect in the action of BC, and to a lesser degree, in the action of OCT. The aim of the present study was to investigate whether the pro-apoptotic action of these drugs involves the increased expression of Bax--a member of Bcl-2 protein family which is known to play an important role in the regulation of apoptosis. Male four-week Fisher 344 rats were used in the experiment. Capsules containing diethylstilboestrol (DES were implanted subcutaneously. Six weeks after the implantation the rats were given OCT (2 x 25 microg/animal/24, BC (3 mg/kg b.w./24 h or OCT and BC at the above doses for 10 days. Bax expression was detected by immunohistochemistry. Prolactin (PRL in blood serum was measured by radioimmunoassay (RIA. It has been found that both OCT and BC, alone or in combination, significantly reduce the tumor weight. Both OCT and BC suppressed PRL levels, but the inhibitory effect of BC was stronger than that of OCT. It has been found that the treatment with OCT and BC, alone or in combination, causes a significant increase in Bax expression in the rat prolactinoma cells. Our findings indicate that anti-tumoral action of bromocriptine and to some extent the action of octreotide in the experimental rat prolactinoma is connected with the induction of apoptosis and is associated with increased Bax expression.

  7. Evaluation of Bax and Bcl-2 Proteins Expression in the Rat Hippocampus due to Childhood Febrile Seizure

    OpenAIRE

    SAEEDI BORUJENI, Mohammad Javad; Hami, Javad; Haghir, Hossein; Rastin, Maryam; Sazegar, Ghasem

    2016-01-01

    Objective Simple Febrile Seizure (SFS) is the most common seizure disorder in childhood, and is frequently described as inoffensive disorder. Nevertheless, there is evidence suggesting the association between neonatal febrile seizures and hippocampal abnormalities in adulthood. This study was conducted at evaluating the hippocampal expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 proteins following SFS induction in rat neonates. Materials & Methods Febrile seizure was modeled by hyper...

  8. Schiff Base Metal Derivatives Enhance the Expression of HSP70 and Suppress BAX Proteins in Prevention of Acute Gastric Lesion

    Directory of Open Access Journals (Sweden)

    Shahram Golbabapour

    2013-01-01

    Full Text Available Schiff base complexes have appeared to be promising in the treatment of different diseases and disorders and have drawn a lot of attention to their biological activities. This study was conducted to evaluate the regulatory effect of Schiff base metal derivatives on the expression of heat shock proteins (HSP 70 and BAX in protection against acute haemorrhagic gastric ulcer in rats. Rats were assigned to 6 groups of 6 rats: the normal control (Tween 20 5% v/v, 5 mL/kg, the positive control (Tween 20 5% v/v, 5 mL/kg, and four Schiff base derivative groups named Schiff_1, Schiff_2, Schiff_3, and Schiff_4 (25 mg/kg. After 1 h, all of the groups received ethanol 95% (5 mL/kg but the normal control received Tween 20 (Tween 20 5% v/v, 5 mL/kg. The animals were euthanized after 60 min and the stomachs were dissected for histology (H&E, immunohistochemistry, and western blot analysis against HSP70 and BAX proteins. The results showed that the Schiff base metal derivatives enhanced the expression of HSP70 and suppressed the expression of BAX proteins during their gastroprotection against ethanol-induced gastric lesion in rats.

  9. Emodin inhibits LOVO colorectal cancer cell proliferation via the regulation of the Bcl-2/Bax ratio and cytochrome c.

    Science.gov (United States)

    Ma, Liang; Li, Wusheng

    2014-10-01

    In this study, the effect of emodin and its mechanism of action were investigated in LOVO colorectal cancer cells. Cell growth was determined using a Cell Counting kit-8 assay, and the results demonstrated that emodin significantly inhibited the growth of LOVO cells in a concentration-dependent manner. In order to investigate the anticancer mechanism of emodin, reverse transcription polymerase chain reaction assays were performed to determine the B-cell lymphoma-2 (Bcl-2)/Bcl-2-associated X protein (Bax) expression ratio in LOVO colorectal cancer cells following treatment with emodin. The results showed that emodin induced a significant increase in the Bax expression level and a marked reduction of the Bcl-2 expression level in LOVO cells. In addition, emodin was found to have an inhibitory effect on the mitochondrial membrane potential and the results from the western blot analysis revealed that cytochrome c was released from the mitochondria to the cytoplasm. In combination, these results suggest that emodin inhibits cancer cell growth via the regulation of the Bcl-2/Bax ratio and by its effect on the mitochondrial apoptosis pathway.

  10. Melatonin promotes Bax sequestration to mitochondria reducing cell susceptibility to apoptosis via the lipoxygenase metabolite 5-hydroxyeicosatetraenoic acid

    KAUST Repository

    Radogna, Flavia

    2015-03-01

    Extra-neurological functions of melatonin include control of the immune system and modulation of apoptosis. We previously showed that melatonin inhibits the intrinsic apoptotic pathway in leukocytes via stimulation of high affinity MT1/MT2 receptors, thereby promoting re-localization of the anti-apoptotic Bcl-2 protein to mitochondria. Here we show that Bcl-2 sequesters pro-apoptotic Bax into mitochondria in an inactive form after melatonin treatment, thus reducing cell propensity to apoptosis. Bax translocation and the anti-apoptotic effect of melatonin are strictly dependent on the presence of Bcl-2, and on the 5-lipoxygenase (5-LOX) metabolite 5-hydroxyeicosatetraenoic acid (5-HETE), which we have previously shown to be produced as a consequence of melatonin binding to its low affinity target calmodulin. Therefore, the anti-apoptotic effect of melatonin requires the simultaneous, independent interaction with high (MT1/MT2) and low (calmodulin) affinity targets, eliciting two independent signal transduction pathways converging into Bax sequestration and inactivation. MT1/MT2 vs. lipoxygenase pathways are activated by 10-9 vs. 10-5M melatonin, respectively; the anti-apoptotic effect of melatonin is achieved at 10-5M, but drops to 10-9M upon addition of exogenous 5-HETE, revealing that lipoxygenase activation is the rate-limiting pathway. Therefore, in areas of inflammation with increased 5-HETE levels, physiological nanomolar concentrations of melatonin may suffice to maintain leukocyte viability.

  11. Preparation and Photocatalytic Properties of Sr2−xBaxTa3O10−yNz Nanosheets

    Directory of Open Access Journals (Sweden)

    Tatsumi Ishihara

    2013-01-01

    Full Text Available Sr2−xBaxTa3O10−yNz (x = 0.0, 0.5, 1.0 nanosheets were prepared by exfoliating layered perovskite compounds (CsSr2−xBaxTa3O10−yNz. The Sr1.5Ba0.5Ta3O9.7N0.2 nanosheet showed the highest photocatalytic activity for H2 production from the water/methanol system among the Sr2−xBaxTa3O9.7N0.2 nanosheets prepared. In addition, Rh-loaded Sr1.5Ba0.5Ta3O9.6N0.3 nanosheet showed the photocatalytic activity for oxygen and hydrogen production from water. The ratio of hydrogen to oxygen evolved was around two. These results indicate that the Rh-loaded Sr1.5Ba0.5Ta3O9.6N0.3 nanosheet is a potential catalyst for photocatalytic water splitting.

  12. Effects of fluoride on liver apoptosis and Bcl-2, Bax protein expression in freshwater teleost, Cyprinus carpio.

    Science.gov (United States)

    Cao, Jinling; Chen, Jianjie; Wang, Jundong; Jia, Ruhui; Xue, Wenjuan; Luo, Yongju; Gan, Xi

    2013-05-01

    Fish take up fluoride directly from water and are the target organisms for fluoride pollution in the aquatic ecosystems. This study was conducted to evaluate oxidative stress, histopathological changes, apoptosis and Bcl-2, Bax expression in the livers of the common carp (Cyprinus carpio) chronically exposed to fluoride. Our results showed that after 90 d of exposure, the inhibition of SOD, GSH activities and a dose-dependent stimulation of MDA levels in the liver tissues indicated that fluoride caused oxidative stress in the fish. Microscopic examinations showed that damages to the liver tissues and cell organelles in the liver tissues increased with exposure concentration. A positive correlation was observed between the apoptosis index and fluoride levels in the livers (r=0.995). There was a negative correlation between the fluoride concentration of water and the expression of Bcl-2, Bcl-2/Bax (r=-0.98, r=-0.96). A positive correlation was showed between the fluoride concentration of water and the expression of Bax (r=0.96) after 90 d of exposure. Our results suggested that the common carp could tolerate relatively high levels of fluoride but adverse effects of fluoride occurred in the livers of the fish after 90 d of exposure. The apoptosis of liver cells had an important causative role in the process of fluoride-induced pathological changes of liver. PMID:23415306

  13. p53's mitochondrial translocation and MOMP action is independent of Puma and Bax and severely disrupts mitochondrial membrane integrity

    Institute of Scientific and Technical Information of China (English)

    Sonja Wolff; Susan Erster; Gustavo Palacios; Ute M Moll

    2008-01-01

    p53's apoptotic program consists of transcription-dependent and transcription-independent pathways. In the latter, physical interactions between mitochondrial p53 and anti-and pro-apoptotic members of the Bcl2 family of mitochondrial permeability regulators are central. Using isogenic cell systems with defined deficiencies, we characterize in detail how mitochondrial p53 contributes to mitochondrial permeabilization, to what extent its action depends on other key Bcl2 family members and define its release activity. We show that mitochondrial p53 is highly efficient in inducing the release of soluble and insoluble apoptogenic factors by severely disrupting outer and inner mitochondrial membrane integrity. This action is associated with wild-type p53-induced oligomerization of Bax, Bak and VDAC and the formation of a stress-induced endogenous complex between p53 and cyclophilin D, normally located at the inner membrane. Tumor-derived p53 mutants are deficient in activating the Bax/Bak lipid pore. These actions are independent of Puma and Bax. Importantly, the latter distinguishes the mitochondrial from the cytosolic p53 death pathway.

  14. Exogenous phosphatidylethanolamine induces apoptosis of human hepatoma HepG2 cells via the bcl-2/bax pathway

    Institute of Scientific and Technical Information of China (English)

    Yu Yao; Chen Huang; Zong-Fang Li; Ai-Ying Wang; Li-Ying Liu; Xiao-Ge Zhao; Yu Luo; Lei Ni; Wang-Gang Zhang; Tu-Sheng Song

    2009-01-01

    AIM: To investigate the signaling pathways implicated in phosphatidylethanolamine (PE)-induced apoptosis of human hepatoma HepG2 cells. METHODS: Inhibitory effects of PE on human hepatoma HepG2 cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell cycle, apoptosis and mitochondrial transmembrane potential (ΔΨm) were analyzed by flow cytometry. Immunocytochemical assay and Western blotting were used to examine Bcl-2, Bax and caspase-3 protein levels in HepG2 cells treated with PE. RESULTS: PE inhibited the growth of HepG2 cells in a dose- and time- dependent manner. It did not affect the cell cycle, but induced apoptosis. PE significantly decreased ΔΨm at 0.25, 0.5 and 1 mmol/L, respectively, suggesting that PE induces cell apoptosis by decreasing the mitochondrial transmembrane potential. The Bcl-2 expression level induced by different concentrations of PE was lower than that in control groups. However, the Bax expression level induced by PE was higher than that in the control group. Meanwhile, PE increased the caspase-3 expression in a dose- and time-dependent manner. CONCLUSION: Exogenous PE induces apoptosis of human hepatoma HepG2 cells via the bcl-2/bax pathway.

  15. Bak compensated for Bax in p53-null cells to release cytochrome c for the initiation of mitochondrial signaling during Withanolide D-induced apoptosis.

    Directory of Open Access Journals (Sweden)

    Susmita Mondal

    Full Text Available The goal of cancer chemotherapy to induce multi-directional apoptosis as targeting a single pathway is unable to decrease all the downstream effect arises from crosstalk. Present study reports that Withanolide D (WithaD, a steroidal lactone isolated from Withania somnifera, induced cellular apoptosis in which mitochondria and p53 were intricately involved. In MOLT-3 and HCT116p53+/+ cells, WithaD induced crosstalk between intrinsic and extrinsic signaling through Bid, whereas in K562 and HCT116p53-/- cells, only intrinsic pathway was activated where Bid remain unaltered. WithaD showed pronounced activation of p53 in cancer cells. Moreover, lowered apoptogenic effect of HCT116p53-/- over HCT116p53+/+ established a strong correlation between WithaD-mediated apoptosis and p53. WithaD induced Bax and Bak upregulation in HCT116p53+/+, whereas increase only Bak expression in HCT116p53-/- cells, which was coordinated with augmented p53 expression. p53 inhibition substantially reduced Bax level and failed to inhibit Bak upregulation in HCT116p53+/+ cells confirming p53-dependent Bax and p53-independent Bak activation. Additionally, in HCT116p53+/+ cells, combined loss of Bax and Bak (HCT116Bax-Bak- reduced WithaD-induced apoptosis and completely blocked cytochrome c release whereas single loss of Bax or Bak (HCT116Bax-Bak+/HCT116Bax+Bak- was only marginally effective after WithaD treatment. In HCT116p53-/- cells, though Bax translocation to mitochondria was abrogated, Bak oligomerization helped the cells to release cytochrome c even before the disruption of mitochondrial membrane potential. WithaD also showed in vitro growth-inhibitory activity against an array of p53 wild type and null cancer cells and K562 xenograft in vivo. Taken together, WithaD elicited apoptosis in malignant cells through Bax/Bak dependent pathway in p53-wild type cells, whereas Bak compensated against loss of Bax in p53-null cells.

  16. Early apoptosis and cell death induced by ATX-S10Na ( Ⅱ)-mediated photodynamic therapy are Bax- and p53-dependent in human colon cancer cells

    Institute of Scientific and Technical Information of China (English)

    Makoto Mitsunaga; Akihito Tsubota; Kohichi Nariai; Yoshihisa Namiki; Makoto Sumi; Tetsuya Yoshikawa; Kiyotaka Fujise

    2007-01-01

    AIM: To investigate the roles of Bax and p53 proteins in photosensitivity of human colon cancer cells by using lysosome-localizing photosensitizer, ATX-S10Na (Ⅱ).METHODS: HCT116 human colon cancer cells and Bax-null or p53-null isogenic derivatives were irradiated with a diode laser. Early apoptosis and cell death in response to photodynamic therapy were determined by MTT assays, annexin V assays, transmission electron microscopy assays, caspase assays and western blotting.RESULTS: Induction of early apoptosis and cell death was Bax- and p53-dependent. Bax and p53 were required for caspase-dependent apoptosis. The levels of anti-apoptotic Bcl-2 family proteins, Bcl-2 and Bcl-XL,were decreased in Bax- and p53-independent manner.CONCLUSION: Our results indicate that early apoptosis and cell death of human colon cancer cells induced by photodynamic therapy with lysosome-localizingphotosensitizer ATX-S10Na (Ⅱ) are mediated by p53-Bax network and Iow levels of Bcl-2 and Bcl-XL proteins.Our results might help in formulating new therapeutic approaches in photedynamic therapy.

  17. In-house validation study of the DuPont Qualicon BAX system Q7 instrument with the BAX system PCR Assay for Salmonella (modification of AOAC Official Method 2003.09 and AOAC Research Institute Performance-Tested Method 100201).

    Science.gov (United States)

    Tice, George; Andaloro, Bridget; White, H Kirk; Bolton, Lance; Wang, Siqun; Davis, Eugene; Wallace, Morgan

    2009-01-01

    In 2006, DuPont Qualicon introduced the BAX system Q7 instrument for use with its assays. To demonstrate the equivalence of the new and old instruments, a validation study was conducted using the BAX system PCR Assay for Salmonella, AOAC Official Method 2003.09, on three food types. The foods were simultaneously analyzed with the BAX system Q7 instrument and either the U.S. Food and Drug Administration Bacteriological Analytical Manual or the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook reference method for detecting Salmonella. Comparable performance between the BAX system and the reference methods was observed. Of the 75 paired samples analyzed, 39 samples were positive by both the BAX system and reference methods, and 36 samples were negative by both the BAX system and reference methods, demonstrating 100% correlation. Inclusivity and exclusivity for the BAX system Q7 instrument were also established by testing 50 Salmonella strains and 20 non-Salmonella isolates. All Salmonella strains returned positive results, and all non-Salmonella isolates returned a negative response. PMID:19610394

  18. 海洛因成瘾模型建立及中脑腹侧被盖区Bax的表达%Effect of heroin on Bax expression in the brain ventral tegmental area and establishment of a rat model of heroin addiction

    Institute of Scientific and Technical Information of China (English)

    俞发荣; 连秀珍; 谢明仁; 张红梅; 李立新; 张琛; 李世平

    2012-01-01

    Objective To study the effects of heroin on Bax expression in the midbrain ventral tegmental area (VTA) of rat. Methods Heroin was intramuscularly injected into rats and a rat model of addiction was generated . Bax expression in the midbrain ventral tegmental area (VTA) was detected by i m muno hist o chemistry. Results The rats had significant withdrawal symptoms after continuous injection of heroin for 7 days. The expression of Bax-positive cells in cere-bellar cortex of rat was significantly increased than that in the control group ( P < 0. 01 ). Conclusion Heroin has effect of inducing Bax gene expression and damage to brain cells .%目的 探索海洛因对中脑腹侧被盖区细胞Bax表达的影响.方法 肌肉注射海洛因,建立成瘾大鼠模型,用免疫组化方法检测中脑腹侧被盖区细胞Bax的表达.结果 连续给大鼠注射海洛因7d后,大鼠出现明显的戒断症状;中脑腹侧被盖区细胞Bax表达阳性细胞比对照组明显增多,与对照组相比差异有显著性 (P<0.01).结论 海洛因具有诱导Bax基因表达、损伤脑组织细胞的作用.

  19. Expression of APE1, Bcl-2 and Bax in retinoblastoma and their clinical significance%APE1、Bcl-2及 Bax在视网膜母细胞瘤中的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    李静; 李德全

    2015-01-01

    目的:分析APE1、Bcl-2及Bax在视网膜母细胞瘤( Rb)中的表达及临床意义。方法选取2011年9月至2013年11月经病理学检查确诊为Rb患者32例及正常视网膜组织16例作为研究对象,免疫组织化学及Western blot分析APE1、Bcl-2及Bax在Rb中及正常视网膜中的表达,比较其在分化及未分化型Rb中的表达。结果 APE1、Bax及Bcl-2在Rb中呈现出高表达,阳性率分别为90.63%、65.63%及68.75%,与正常组比差异均具有统计学意义(χ2=30.13,χ2=12.31,χ2=16.91, P <0.01),与Western blot结果一致;分化组与未分化组中APE1、Bax存在差异显著(χ2=4.99,χ2=7.85, P <0.05),Bcl-2无统计学差异(χ2=0.73, P >0.01)。结论 Rb的发生发展涉及多个基因及生物学过程,分析APE1、bcl-2及bax在Rb中的表达,对Rb的诊断与治疗有重要的参考价值。%Objective To analyze the expression of APE1, Bcl-2 and Bax in retinoblastoma (Rb) and to evaluate their clinical significance.Methods A total of 32 retinoblastoma patients were enrolled for this study from September 2011 to November 2013.Sixteen normal retinal tissues were collected as control.Immunohistochemistry and Western blot were used to evaluate the expression of APE1, Bcl-2 and Bax in retinoblastoma tumor tissues and normal retina.Their expres-sions in differentiated and undifferentiated Rb were also compared.Results APE1, Bax and Bcl-2 were highly expressed in retinoblastoma with positive rates being 90.63%, 65.63% and 68.75%, respectively, and were significantly higher than in normal retina tissues (χ2 =30.13 for APE1,χ2 =12.31 for Bax, andχ2 =16.91 for Bcl-2;P 0.01).Conclusion The development of Rb involves multiple genes and biological processes.Analysis of the expression of APE1, Bcl-2 and Bax in Rb has important clinical value for the diagnosis and treat-ment of Rb.

  20. Engineering of the Curie temperature of epitaxial Sr1-xBaxTiO3 films via strain

    Science.gov (United States)

    Dai, Y.; Schubert, J.; Hollmann, E.; Mussler, G.; Wördenweber, R.

    2016-09-01

    The impact of strain on the structural and electrical properties of epitaxial Sr1-xBaxTiO3 films grown on single crystalline DyScO3 (110), TbScO3 (110), and GdScO3 (110) substrates is presented. X-ray diffraction measurements demonstrate that all films are grown epitaxially. The tensile in-plane strain is only partially compensated by a contraction of the out-of-plane lattice parameter. As a result, the volume of the unit cell of the Sr1-xBaxTiO3 film increases due to the tensile strain, and the resulting Poisson ratio of the film is ν ≈ 0.33, which is larger than but still close to the literature values of ν ≈ 0.23 for unstrained defect-free SrTiO3. The Curie temperature derived from the temperature dependence of the in-plane dielectric response leads to a strain-temperature phase diagram for the epitaxial Sr1-xBaxTiO3 films. The experimental data show a deviation from the linear dependence predicted by the Landau thermodynamic theory for large strain (>1.2%). However, using the equilibrium thermodynamic analysis, we can demonstrate that this deviation arises from the relaxation of the strain due to defect formation in the film. The result reveals that in addition to the nominal misfit strain, the defect formation strongly affects the effective strain and, thus, the dielectric response of epitaxially grown ferroelectric films.

  1. First Principles Calculations of Structural, Electronic, Thermodynamic and Thermal Properties of BaxSr1-xTe Ternary Alloys

    Science.gov (United States)

    Chelli, S.; Meradji, H.; Amara Korba, S.; Ghemid, S.; El Haj Hassan, F.

    2014-12-01

    The structural, electronic thermodynamic and thermal properties of BaxSr1-xTe ternary mixed crystals have been studied using the ab initio full-potential linearized augmented plane wave (FP-LAPW) method within density functional theory (DFT). In this approach, the Perdew-Burke-Ernzerhof-generalized gradient approximation (PBE-GGA) was used for the exchange-correlation potential. Moreover, the recently proposed modified Becke Johnson (mBJ) potential approximation, which successfully corrects the band-gap problem was also used for band structure calculations. The ground-state properties are determined for the cubic bulk materials BaTe, SrTe and their mixed crystals at various concentrations (x = 0.25, 0.5 and 0.75). The effect of composition on lattice constant, bulk modulus and band gap was analyzed. Deviation of the lattice constant from Vegard's law and the bulk modulus from linear concentration dependence (LCD) were observed for the ternary BaxSr1-xTe alloys. The microscopic origins of the gap bowing were explained by using the approach of Zunger and co-workers. On the other hand, the thermodynamic stability of these alloys was investigated by calculating the excess enthalpy of mixing, ΔHm as well as the phase diagram. It was shown that these alloys are stable at high temperature. Thermal effects on some macroscopic properties of BaxSr1-xTe alloys were investigated using the quasi-harmonic Debye model, in which the phononic effects are considered.

  2. Intermittent hypoxia attenuates ischemia/reperfusion induced apoptosis in cardiac myocytes via regulating Bcl-2/Bax expression

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Intermittent hypoxia has been shown to provide myocardial protection against ishemia/reperfusion-induced injury.Cardiac myocyte loss through apoptosis has been reported in ischemia/reperfusion injury. Our aim was to investigate whether intermittent hypoxia could attenuate ischemia/reperfusion-induced apoptosis in cardiac myocytes and its potential mechanisms. Adult male Sprague-Dawley rats were exposed to hypoxia simulated 5000 m in a hypobaric chamber for 6 h/day, lasting 42 days. Normoxia group rats were kept under normoxic conditions. Isolated perfused hearts from both groups were subjected to 30 min of global ischemia followed by 60 min reperfusion.Incidence of apoptosis in cardiac myocytes was determined by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and DNA agarose gel electrophoresis. Expressions of apoptosis related proteins,Bax and Bcl-2, in cytosolic and membrane fraction were detected by Western Blotting. After ischemia/reperfusion,enhanced recovery of cardiac function was observed in intermittent hypoxia hearts compared with normoxia group.Ischemia/reperfusion-induced apoptosis, as evidenced by TUNEL-positive nuclei and DNA fragmentation, was significantly reduced in intermittent hypoxia group compared with normoxia group. After ischemia/reperfusion,expression of Bax in both cytosolic and membrane fractions was decreased in intermittent hypoxia hearts compared with normoxia group. Although ischemia/reperfusion did not induce changes in the level of Bcl-2 expression in cytosolic fraction between intermittent hypoxia and normoxia groups, the expression of Bcl-2 in membrane fraction was upregulated in intermittent hypoxia group compared with normoxia group. These results indicated that the cardioprotection of intermittent hypoxia against ischemia/reperfusion injury appears to be in part due to reduce myocardial apoptosis. Intermittent hypoxia attenuated ischemia/reperfusion-induced apoptosis via increasing the ratio of Bcl

  3. Involvement of P53 and Bax/Bad triggering apoptosis in thioacetamide-induced hepatic epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Li-Hsuen Chen; Chia-Yu Hsu; Ching-Feng Weng

    2006-01-01

    AIM: Thioacetamide (TAA) has been used in studying liver fibrosis and cirrhosis, however, the mechanisms of TAA-induced apoptosis in liver are still unclear. The hepatic epithelial cell line clone 9 was cultured and treated with TAA to investigate the causes of cell death. METHODS: The cell viability of TAA-induced clone 9 cells was determined using MTT assay. Total cellular GSH in TAA-induced clone 9 cells was measured using a slight modification of the Tietze assay. The activity of caspase 3 in TAA-induced clone 9 cells was monitored by the cleavage of DEVD-p-nitroanaline. TUNEL assay and flow cytometry were applied for the determination of DNA fragmentation and the proportion of apoptosis in TAAinduced clone 9 cells, respectively. The alterations of caspase 3, Bad, Bax and Phospho-P53 contents in TAAinduced clone 9 cells were measured by Western blot. RESULTS: The experimental data indicated that TAA caused rat hepatic epithelial cell line clone 9 cell death in a dose-and time-dependent manner; 60% of the cells died (MTT assay) within 24 h after 100 mg/L TAA was applied. Apoptotic cell percentage (TUNEL assay) and caspase 3 activities were highest after 100 mg/L TAA was added for 8 h. The release of GSH and the elevation in caspase content after TAA treatment resulted in clone 9 cell apoptosis via oxidative stress and a caspasedependent mechanism. The phospho-p53, Bax and Bad protein expressions in clone 9 cells were increased after TAA treatment.CONCLUSION: These results reveal that TAA activates p53, increases caspase 3, Bax and Bad protein contents,perhaps causing the release of cytochrome c from mitochondria and the disintegration of membranes, leading to apoptosis of cells.

  4. Downregulation of uPAR and cathepsin B induces apoptosis via regulation of Bcl-2 and Bax and inhibition of the PI3K/Akt pathway in gliomas.

    Directory of Open Access Journals (Sweden)

    Ramarao Malla

    Full Text Available BACKGROUND: Glioma is the most commonly diagnosed primary brain tumor and is characterized by invasive and infiltrative behavior. uPAR and cathepsin B are known to be overexpressed in high-grade gliomas and are strongly correlated with invasive cancer phenotypes. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we observed that simultaneous downregulation of uPAR and cathepsin B induces upregulation of some pro-apoptotic genes and suppression of anti-apoptotic genes in human glioma cells. uPAR and cathepsin B (pCU-downregulated cells exhibited decreases in the Bcl-2/Bax ratio and initiated the collapse of mitochondrial membrane potential. We also observed that the broad caspase inhibitor, Z-Asp-2, 6-dichlorobenzoylmethylketone rescued pCU-induced apoptosis in U251 cells but not in 5310 cells. Immunoblot analysis of caspase-9 immunoprecipitates for Apaf-1 showed that uPAR and cathepsin B knockdown activated apoptosome complex formation in U251 cells. Downregulation of uPAR and cathepsin B also retarded nuclear translocation and interfered with DNA binding activity of CREB in both U251 and 5310 cells. Further western blotting analysis demonstrated that downregulation of uPAR and cathepsin B significantly decreased expression of the signaling molecules p-PDGFR-β, p-PI3K and p-Akt. An increase in the number of TUNEL-positive cells, increased Bax expression, and decreased Bcl-2 expression in nude mice brain tumor sections and brain tissue lysates confirm our in vitro results. CONCLUSIONS/SIGNIFICANCE: In conclusion, RNAi-mediated downregulation of uPAR and cathepsin B initiates caspase-dependent mitochondrial apoptosis in U251 cells and caspase-independent mitochondrial apoptosis in 5310 cells. Thus, targeting uPAR and cathepsin B-mediated signaling using siRNA may serve as a novel therapeutic strategy for the treatment of gliomas.

  5. On the moment of inertia in deformed Ba-Xe nuclei as deduced from gamma-gamma energy correlation experiments

    International Nuclear Information System (INIS)

    The γ-rays following reactions induced by bombarding targets of 114,116,118,120,122Sn with 118 MeV 12C ions were investigated using six NaI(Tl) detectors in a two-dimensional coincidence arrangement. Experimental energy-correlation spectra were extracted from the original coincidence matrices. The energy-correlation spectra exhibit the features expected for rotational nuclei and were used to deduce information on the moment of inertia Isup((2)) = ΔI/Δω. The gross properties of the behaviour of Isup((2)) in the Ba-Xe region are discussed together with their interpretation within the cranked shell model (CSM). (orig.)

  6. Retinoids cause apoptosis in pancreatic cancer cells via activation of RAR-γ and altered expression of Bcl-2/Bax

    OpenAIRE

    Pettersson, F; Dalgleish, A G; Bissonnette, R P; Colston, K W

    2002-01-01

    All-trans-retinoic acid and 9-cis-retinoic acid have been reported to have inhibitory effects on pancreatic adenocarcinoma cells and we have shown that this is partly due to induction of apoptosis. In this study, the mechanisms whereby 9-cis-retinoic acid induces apoptosis in these cells were investigated. An involvement of the Bcl-2 family of proteins was shown, such that 9-cis-retinoic acid causes a decrease in the Bcl-2/Bax ratio. Overexpression of Bcl-2 also resulted in inhibition of apop...

  7. The protective role of Bax Inhibitor-1 against chronic mild stress through the inhibition of monoamine oxidase A

    OpenAIRE

    Hwa-Young Lee; Geum-Hwa Lee; Anu Marahatta; Shun-Mei Lin; Mi-Rin Lee; Kyu Yun Jang; Kyung Min Kim; Hee Jae Lee; Jae-Won Lee; Tarique Rajasaheb Bagalkot; Young-Chul Chung; Yong-Chul Lee; Hyung-Ryong Kim; Han-Jung Chae

    2013-01-01

    The anti-apoptotic protein Bax inhibitor-1 (BI-1) is a regulator of apoptosis linked to endoplasmic reticulum (ER) stress. It has been hypothesized that BI-1 protects against neuron degenerative diseases. In this study, BI-1−/− mice showed increased vulnerability to chronic mild stress accompanied by alterations in the size and morphology of the hippocampi, enhanced ROS accumulation and an ER stress response compared with BI-1+/+ mice. BI-1−/− mice exposed to chronic mild stress showed signif...

  8. BaP-induced DNA damage initiated p53-independent necroptosis via the mitochondrial pathway involving Bax and Bcl-2.

    Science.gov (United States)

    Jiang, Y; Chen, X; Yang, G; Wang, Q; Wang, J; Xiong, W; Yuan, J

    2013-12-01

    Benzo(a)pyrene (BaP), a typical environmental carcinogen, can induce cell death both by protein 53 or tumor protein 53 (p53)-independent and -dependent pathways. However, little is known about the molecular mechanisms of p53-independent pathways in BaP-induced cell death. In this study, cells with different genetic background (including p53-proficient human fetal lung fibroblast cell lines (MRC-5), p53-deficient human non-small-cell lung carcinoma cell lines (H1299), and p53-knockdown cell lines (MRC-5(p53-/-))) were used to establish models of BaP-induced cell death. The results showed that BaP (8, 16, 32, and 64 μM) induced necroptotic cell death in the cell lines. The necroptotic cell death and DNA damage were concurrently observed. In the three cell lines, at 24 h after treatment, BaP (8-64 μM) upregulated expressions of BAX, BCL-2, and cleaved caspase-3 proteins, but not their messenger RNA levels. The findings suggested that BaP-induced necroptosis was modulated by the p53-independent pathway, which was related to the induction of BAX, decreased expression of BCL-2, and activation of caspase-3.

  9. Bcl-2, Bax, and c-Fos expression correlates to RPE cell apoptosis induced by UV-light and daunorubicin

    DEFF Research Database (Denmark)

    Liang, Y G; Jorgensen, A G; Kaestel, C G;

    2000-01-01

    PURPOSE. The aim of this study was to determine the role of Bcl-2, Bcl-X L, Bax, and c-Fos in regulation of apoptosis, induced by ultraviolet-light A (UV-A) and daunorubicin (DNR), in retinal pigment epithelium (RPE) cells grown on bovine extracellular matrix (ECM)-coated or uncoated plastic dishes....... METHODS. Apoptosis in confluent RPE cells cultured on ECM-coated or uncoated dishes was induced by UV-A or DNR. Apoptosis was detected by 7-amino-actinomycin D labeling followed by flow cytometry and by terminal deoxy-transferase mediated X-dUTP nick end labeling (TUNEL). Cellular expression of Bcl-2, Bcl......-X L, Bax, and c-Fos was determined by the use of antibodies and flow cytometry, Western blot analysis, and immunocytochemical staining. RESULTS. Both UV-A and DNR induce apoptosis in human RPE cells in vitro. Human fetal RPE cells grown on ECM-coated dishes were significantly more resistant to UV...

  10. Expression of p53, Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease

    Institute of Scientific and Technical Information of China (English)

    Anatol Panasiuk; Janusz Dzieciol; Bozena Panasiuk; Danuta Prokopowicz

    2006-01-01

    AIM: To analyze the protein expression essential for apoptosis in liver steatosis.METHODS: The expression of proapoptotic proteinsp53, Bax, and antiapoptotic Bcl-2 in hepatocytes with steatosis (SH) and without steatosis (NSH) was evaluated in 84 patients at various stages of non-alcoholic fatty liver disease (NAFLD).RESULTS: Immunohistochemical staining of liver tissue showed the activation of p53 protein in SH and NSH with increased liver steatosis, diminished Bcl-2 and slightly decreased Bax protein. Positive correlation was found between the stage of liver steatosis with p53 expression in SH (r = 0.54, P < 0.01) and NSH (r = 0.49,P < 0.01).The antiapoptotic protein Bcl-2 was diminished together with the advancement of liver steatosis, especially in non-steatosed hepatocytes (r =0.43, P < 001).CONCLUSION: Apoptosis is one of the most important mechanisms leading to hepatocyte elimination in NAFLD. The intensification of inflammation in NAFLD induces proapoptotic protein p53 with the inhibition of antiapoptotic Bcl-2.

  11. (-)-Anonaine induces apoptosis through Bax- and caspase-dependent pathways in human cervical cancer (HeLa) cells.

    Science.gov (United States)

    Chen, Chung-Yi; Liu, Tsan-Zon; Tseng, Wei-Chang; Lu, Fung-Jou; Hung, Ray-Ping; Chen, Chi-Hung; Chen, Ching-Hsein

    2008-08-01

    (-)-Anonaine has been shown to have some anticancer activities, but the mechanisms of (-)-anonaine inducing cell death of human cancer cells is not fully understood. We investigated the mechanisms of apoptosis induced by (-)-anonaine in human HeLa cancer cells. Treatment with (-)-anonaine induces dose-dependent DNA damage that is correlated with increased intracellular nitric oxide, reactive oxygen species, glutathione depletion, disruptive mitochondrial transmembrane potential, activation of caspase 3, 7, 8, and 9, and poly ADP ribose polymerase cleavage. Our data indicate that (-)-anonaine up-regulated the expression of Bax and p53 proteins in HeLa cancer cells. The apoptosis and expression of Bax induced by (-)-anonaine could be inhibited when the HeLa cells were pretreated with Boc-Asp(OMe)-fmk, which is a broad caspases inhibitor. There was no obvious DNA damage in the (-)-anonaine-treated Madin-Darby canine kidney and Vero cell lines. Both Madin-Darby canine kidney and Vero cell lines are kidney epithelial cellular morphology. These results suggest that (-)-anonaine might be considered a potent compound for chemotherapy against cervical cancer or a health food supplement for cancer chemoprevention.

  12. Yeast Bax inhibitor, Bxi1p, is an ER-localized protein that links the unfolded protein response and programmed cell death in Saccharomyces cerevisiae.

    Directory of Open Access Journals (Sweden)

    James Cebulski

    Full Text Available Bax inhibitor-1 (BI-1 is an anti-apoptotic gene whose expression is upregulated in a wide range of human cancers. Studies in both mammalian and plant cells suggest that the BI-1 protein resides in the endoplasmic reticulum and is involved in the unfolded protein response (UPR that is triggered by ER stress. It is thought to act via a mechanism involving altered calcium dynamics. In this paper, we provide evidence that the Saccharomyces cerevisiae protein encoded by the open reading frame, YNL305C, is a bona fide homolog for BI-1. First, we confirm that yeast cells from two different strain backgrounds lacking YNL305C, which we have renamed BXI1, are more sensitive to heat-shock induced cell death than wildtype controls even though they have indistinguishable growth rates at 30°C. They are also more susceptible both to ethanol-induced and to glucose-induced programmed cell death. Significantly, we show that Bxi1p-GFP colocalizes with the ER localized protein Sec63p-RFP. We have also discovered that Δbxi1 cells are not only more sensitive to drugs that induce ER stress, but also have a decreased unfolded protein response as measured with a UPRE-lacZ reporter. Finally, we have discovered that deleting BXI1 diminishes the calcium signaling response in response to the accumulation of unfolded proteins in the ER as measured by a calcineurin-dependent CDRE-lacZ reporter. In toto, our data suggests that the Bxi1p, like its metazoan homologs, is an ER-localized protein that links the unfolded protein response and programmed cell death.

  13. JNK-Bcl-2/Bcl-xL-Bax/Bak Pathway Mediates the Crosstalk between Matrine-Induced Autophagy and Apoptosis via Interplay with Beclin 1

    Directory of Open Access Journals (Sweden)

    Jiong Yang

    2015-10-01

    Full Text Available Autophagy is associated with drug resistance which has been a threat in chemotherapy of hepatocellular carcinoma (HCC. The interconnected molecular regulators between autophagy and apoptosis serve as switching points critical to the ultimate outcome of the cell. Our study was performed to investigate the crosstalk between autophagy and apoptosis in HCC after the treatment of matrine. Flow cytometry and TUNEL (terminal dexynucleotidyl transferase (TdT-mediated dUTP nick end labeling assay were used to detect apoptosis in vitro and in vivo, respectively. Bax oligomerization and Cytochrome c release assay were performed. Immunoprecipitation and siRNA transfection were used to detect the interplay between Bcl-2/Bcl-xL,Bax, and Beclin 1. Our results showed that: (1 matrine not only activated caspase and PARP (poly ADP-ribose polymerase cleavage, but also triggered autophagy as shown by the increased levels of LC3II, Beclin 1, and PI3KC3, and the decreased level of p62; (2 matrine treatment promoted the JNK-Bcl-2/ Bcl-xL-Bax/Bak pathway; (3 Bax was oligomerized, the mitochondrial membrane potential altered, and Cytochrome c was released subsequently; (4 Bax interacts with Beclin 1 and inhibits autophagy, which may be a new crosstalk point; and (5 finally, we showed that matrine suppressed the growth of a MHCC97L xenograft in vivo for the first time. In conclusion, the JNK-Bcl-2/Bcl-xL-Bax/Bak pathway mediates the crosstalk between matrine-induced autophagy and apoptosis via interplay with Beclin 1.

  14. JNK-Bcl-2/Bcl-xL-Bax/Bak Pathway Mediates the Crosstalk between Matrine-Induced Autophagy and Apoptosis via Interplay with Beclin 1.

    Science.gov (United States)

    Yang, Jiong; Yao, Shukun

    2015-10-27

    Autophagy is associated with drug resistance which has been a threat in chemotherapy of hepatocellular carcinoma (HCC). The interconnected molecular regulators between autophagy and apoptosis serve as switching points critical to the ultimate outcome of the cell. Our study was performed to investigate the crosstalk between autophagy and apoptosis in HCC after the treatment of matrine. Flow cytometry and TUNEL (terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling) assay were used to detect apoptosis in vitro and in vivo, respectively. Bax oligomerization and Cytochrome c release assay were performed. Immunoprecipitation and siRNA transfection were used to detect the interplay between Bcl-2/Bcl-xL,Bax, and Beclin 1. Our results showed that: (1) matrine not only activated caspase and PARP (poly ADP-ribose polymerase) cleavage, but also triggered autophagy as shown by the increased levels of LC3II, Beclin 1, and PI3KC3, and the decreased level of p62; (2) matrine treatment promoted the JNK-Bcl-2/ Bcl-xL-Bax/Bak pathway; (3) Bax was oligomerized, the mitochondrial membrane potential altered, and Cytochrome c was released subsequently; (4) Bax interacts with Beclin 1 and inhibits autophagy, which may be a new crosstalk point; and (5) finally, we showed that matrine suppressed the growth of a MHCC97L xenograft in vivo for the first time. In conclusion, the JNK-Bcl-2/Bcl-xL-Bax/Bak pathway mediates the crosstalk between matrine-induced autophagy and apoptosis via interplay with Beclin 1.

  15. Effects of L-Tetrahydropalmatine on the Expressions of bcl-2 and bax in Rat after Acute Global Cerebral Ischemia and Reperfusion

    Institute of Scientific and Technical Information of China (English)

    刘彬; 杨光田

    2004-01-01

    To investigate the effects of L-Tetrahydropalmatine (L-THP) on the expressions of bcl2, bax and neuronal apoptosis after cerebral ischemia and reperfusion, 60 Wistars rats were randomly divided into 3 groups: sham-operation group (group S, n = 20), ischemic-reperfusion group treated with saline (group I, n=20) and ischemia-reperfusion group treated with L-THP (group T, n=20) . The rat model of global cerebral ischemia and reperfusion was induced by Pulsinelli's four-vessel occlusion method. The expression of bcl-2 and bax mRNA was detected by in situ hybridization and reverse transcriptional polymerase chain reaction (RT-PCR). The number of apoptotic neurons was examined by terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method. Compared with group S, the expression of bcl-2 and bax mRNA in group I was increased significantly (P<0.01), and the number of apoptotic neurons increased either (P<0.01). After L-THP treatment, the expression of bcl-2 mRNA was up-regulated (P<0.01) and that of bax mRNA was down-regulated (P<0.01); the number of apoptotic neurons was decreased (P<0.01). Our results indicated that bcl-2 may suppress apoptosis and bax promote apoptosis after cerebral ischemia and reperfusion. L-THP could ameliorate cerebral ischemia and reperfusion damage by reducing the apoptosis through regulating bcl-2 and bax.

  16. Apoptosis and the activity of ceramide, Bax and Bcl-2 in the lungs of neonatal rats exposed to limited and prolonged hyperoxia

    Directory of Open Access Journals (Sweden)

    Bitar Fadi F

    2006-07-01

    Full Text Available Abstract Background The aim of the study is to examine the effect of limited and prolonged hyperoxia on neonatal rat lung. This is done by examining the morphologic changes of apoptosis, the expression of ceramide, an important mediator of apoptosis, the expression of inflammatory mediators represented by IL-1β and the expression of 2 proto-oncogenes that appear to modulate apoptosis (Bax and Bcl-2. Methods Newborn rats were placed in chambers containing room air or oxygen above 90% for 7 days. The rats were sacrificed at 3, 7 or 14 days and their lungs removed. Sections were fixed, subjected to TUNEL, Hoechst, and E-Cadherin Staining. Sections were also incubated with anti-Bcl-2 and anti-Bax antisera. Bcl-2 and Bax were quantitated by immunohistochemistry. Lipids were extracted, and ceramide measured through a modified diacylglycerol kinase assay. RT-PCR was utilized to assess IL-1β expression. Results TUNEL staining showed significant apoptosis in the hyperoxia-exposed lungs at 3 days only. Co-staining of the apoptotic cells with Hoechst, and E-Cadherin indicated that apoptotic cells were mainly epithelial cells. The expression of Bax and ceramide was significantly higher in the hyperoxia-exposed lungs at 3 and 14 days of age, but not at 7 days. Bcl-2 was significantly elevated in the hyperoxia-exposed lungs at 3 and 14 days. IL-1β expression was significantly increased at 14 days. Conclusion Exposure of neonatal rat lung to hyperoxia results in early apoptosis documented by TUNEL assay. The early rise in Bax and ceramide appears to overcome the anti-apoptotic activity of Bcl-2. Further exposure did not result in late apoptotic changes. This suggests that apoptotic response to hyperoxia is time sensitive. Prolonged hyperoxia results in acute lung injury and the shifting balance of ceramide, Bax and Bcl-2 may be related to the evolution of the inflammatory process.

  17. 深Ⅱ度烧伤愈合后Bcl-xl、Bax蛋白表达与增生性瘢痕的相关性%The relationship of hypertrophic scar with the expression of Bcl-xl and Bax proteins in deep Ⅱ burns coalesce skin

    Institute of Scientific and Technical Information of China (English)

    李泰然; 王振宇; 孟庆延; 骆宁

    2012-01-01

    目的 探讨Bcl-xl和Bax蛋白在深Ⅱ度烧伤愈合后不同时期增生性瘢痕中的表达特点.方法 人深Ⅱ度烧伤愈合后不同时期的增生性瘢痕皮肤40例,正常皮肤组织10例.分为增生期组、减退早期组、减退晚期组、成熟期组,正常对照组.免疫组织化学染色,检测Bcl-xl和Bax蛋白的表达.结果 深Ⅱ度烧伤创面愈合后,Bcl-xl和Bax蛋白主要表达于表皮基底细胞和真皮层成纤维细胞,增生期和减退早期Bcl-xl和Bax蛋白产物光密度值明显高于正常皮肤(P<0.01),减退晚期和成熟期随瘢痕成熟而逐渐递减接近正常皮肤.结论 Bcl-xl和Bax蛋白表达与深Ⅱ度烧伤创面愈合后增生性瘢痕的发生和瘢痕成熟相关.%Objective To study the expression and histological distribution of apoptosis-related gene Bel-xl and Bax in hypertrophic scars (HS) in deep Ⅱ burns coalesce skin. Method Ten specimens of normal skin (NS), 40 skin specimens of hypertrophic scars (HS) in deep II bums coalesce skin were divided into four groups: hypertrophic phase group (1-6 months after burned), early hypotrophic group (7-12 months after burned), later hypotrophic group (13-18 months after burned) and full recovered stage group (longer than 18 months). Immunochemistry was used to detect the expressions of Bcl-xl and Bax protein. Results The Bcl-xl and Bax positive product was found mainly in the fibroblast cytoplasm and nucleus of the basal layer in normal skin and deep II burns coalesce skin. The average optical density values of Bcl-xl and Bax protein product were significantly higher in the hypertrophic phase group and early hypotrophic group than in control group (P< 0.01), but decreased gradually to normal skin in later hypotrophic group and full recovered stage group. Conclusion The expressions of Bcl-xl and Bax proteins are related to the occurrence of hvpertrophic scars and wound healing in deep II burns coalesce skin.

  18. Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and -independent mechanisms and contributes to drug resistance

    DEFF Research Database (Denmark)

    Erler, Janine Terra; Cawthorne, Christopher J; Williams, Kaye J;

    2004-01-01

    of the Bcl-2 protein family. Oxygen deprivation of human colon cancer cells in vitro provoked decreased mRNA and protein levels of proapoptotic Bid and Bad. Hypoxia-inducible factor 1 (HIF-1) was dispensable for the down-regulation of Bad but required for that of Bid, consistent with the binding of HIF-1......alpha to a hypoxia-responsive element (positions -8484 to -8475) in the bid promoter. Oxygen deprivation resulted in proteosome-independent decreased expression of Bax in vitro, consistent with a reduction in global translation efficiency. The physiological relevance of Bid and Bax down...

  19. Determination of stoichiometry and concentration of trace elements in thin BaxSr1-xTiO3 perovskite layers.

    Science.gov (United States)

    Becker, J S; Boulyga, S F

    2001-07-01

    This paper describes an analytical procedure for determining the stoichiometry of BaxSr1-xTiO3 perovskite layers using inductively coupled plasma mass spectrometry (ICP-MS). The analytical results of mass spectrometry measurements are compared to those of X-ray fluorescence analysis (XRF). The performance and the limits of solid-state mass spectrometry analytical methods for the surface analysis of thin BaxSr1-xTiO3 perovskite layers sputtered neutral mass spectrometry (SNMS)--are investigated and discussed. PMID:11496982

  20. Immunohistochemical expression of p53, BCL-2, BAX and VEGFR1 proteins in nephroblastomas A expressão imuno-histoquímica das proteínas p53, BCL-2, BAX e VEGFR1 em nefroblastomas

    Directory of Open Access Journals (Sweden)

    Ana Paula Percicote

    2013-02-01

    Full Text Available INTRODUCTION: Nephroblastoma or Wilms' tumor is the most frequent renal cancer in children. Although its prognosis is favorable for most patients, it may relapse or have a fatal outcome. The characterization of risk groups by applying immunohistochemical biomarkers aims to adapt the treatment to its corresponding group as well as to reduce relapses and fatal outcome. p53, B-cell lymphoma 2 (BCL-2, BCL-2 associated protein X (BAX and vascular endothelial growth factor receptor 1 (VEGFR1 are among the most widely studied biomarkers, which are related to the apoptotic pathway, DNA repair and neovascularization. OBJECTIVE: The objective of this study is to assess the immunohistochemical expression of p53, BCL-2, BAX and VEGFR1 in samples of human nephroblastoma and to correlate them with clinicopathological prognostic factors. MATERIAL AND METHODS: Twenty-nine surgical specimens of nephroblastoma diagnosed from 1994 to 2007 were selected from the Anatomopathological Service of two hospitals in Curitiba. The immunohistochemical analysis of tissue microarrays was performed through immunoperoxidase staining and the yielded results were compared with clinicopathological prognostic factors. RESULTS: The major immunohistochemical expression of VEGFR1 in blastema and epithelium presented positive association with the risk group. Hence this may be related to higher vascular neoplastic invasion apparently caused by the endothelial growth factor, which maximizes the chances of metastasis and ultimately changes tumor staging, risk group and clinical evolution. CONCLUSIONS: The immunohistochemical expression of VEGFR1 substantiated a directly proportional association with the nephroblastoma risk group.INTRODUÇÃO: O nefroblastoma, ou tumor de Wilms, é a neoplasia renal mais frequente na infância. Embora o prognóstico seja favorável para a maioria dos pacientes, muitos evoluem para recidiva ou óbito. A caracterização de grupos de risco por meio de

  1. Curcuma purpurascens BI. rhizome accelerates rat excisional wound healing: involvement of Hsp70/Bax proteins, antioxidant defense, and angiogenesis activity

    Directory of Open Access Journals (Sweden)

    Rouhollahi E

    2015-10-01

    Full Text Available Elham Rouhollahi,1 Soheil Zorofchian Moghadamtousi,2 Fatemeh Hajiaghaalipour,3 Maryam Zahedifard,2 Faezeh Tayeby,2 Khalijah Awang,4 Mahmood Ameen Abdulla,3 Zahurin Mohamed1 1Pharmacogenomics Laboratory, Department of Pharmacology, Faculty of Medicine, 2Institute of Biological Sciences, Faculty of Science, 3Department of Biomedical Science, Faculty of Medicine, 4Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia Purpose: Curcuma purpurascens BI. is a member of Zingiberaceae family. The purpose of this study is to investigate the wound healing properties of hexane extract of C. purpurascens rhizome (HECP against excisional wound healing in rats.Materials and methods: Twenty four rats were randomly divided into 4 groups: A negative control (blank placebo, acacia gum, B low dose of HECP, C high dose of HECP, and D positive control, with 6 rats in each group. Full-thickness incisions (approximately 2.00 cm were made on the neck area of each rat. Groups 1–4 were treated two-times a day for 20 days with blank placebo, HECP (100 mg/kg, HECP (200 mg/kg, and intrasite gel as a positive control, respectively. After 20 days, hematoxylin and eosin and Masson’s trichrome stainings were employed to investigate the histopathological alterations. Protein expressions of Bax and Hsp70 were examined in the wound tissues using immunohistochemistry analysis. In addition, levels of enzymatic antioxidants and malondialdehyde representing lipid peroxidation were measured in wound tissue homogenates.Results: Macroscopic evaluation of wounds showed conspicuous elevation in wound contraction after topical administration of HECP at both doses. Moreover, histopathological analysis revealed noteworthy reduction in the scar width correlated with the enhanced collagen content and fibroblast cells, accompanied by a reduction of inflammatory cells in the granulation tissues. At the molecular level, HECP facilitates wound-healing process

  2. Study of BaxSr1-xTiO3 thin films using transverse-field Ising model

    Institute of Scientific and Technical Information of China (English)

    Tao Yong-Mei; Jiang Qing

    2004-01-01

    In this paper, the effects of doping on the thermodynamic properties of BaxSr1-xTiO3 (BST) thin film are investigated, based on the transverse-field Ising model (TIM) within the framework of mean field theory. We apply the double-peak distribution model of related parameters to mimic doping. The lattice expansion arising from doping with large Ba2+ was also taken into account. We concentrate on the doping concentration dependence of peak temperature (Tm), spontaneous polarization and dielectric susceptibility. It is found that the doping concentration has great influence on the dielectric properties and phase transition properties of BST thin films. We also discuss the quantum effect arising from doping.

  3. Effects of aspartame on hsp70, bcl-2 and bax expression in immune organs of Wistar albino rats

    Science.gov (United States)

    Choudhary, Arbind Kumar; Devi, Rathinasamy Sheela

    2016-01-01

    Abstract Aspartame, a “first generation sweetener”, is widely used in a variety of foods, beverages, and medicine. The FDA has determined the acceptable daily intake (ADI) value of aspartame to be 50 mg/kg·day, while the JECFA (Joint FAO/WHO Expert Committee on Food Additives) has set this value at 40 mg/kg of body weight/day. Safety issues have been raised about aspartame due to its metabolites, specifically toxicity from methanol and/or its systemic metabolites formaldehyde and formic acid. The immune system is now recognized as a target organ for many xenobiotics, such as drugs and chemicals, which are able to trigger unwanted apoptosis or to alter the regulation of apoptosis. Our previous studies has shown that oral administration of aspartame [40 mg/(kg·day)] or its metabolites for 90 days increased oxidative stress in immune organs of Wistar albino rats. In this present study, we aimed to clarify whether aspartame consumption over a longer period (90-days) has any effect on the expression of hsp70, bcl-2 and bax at both mRNA transcript and protein expression levels in immune organs. We observed that oral administration of aspartame for 90 days did not cause any apparent DNA fragmentation in immune organs of aspartame treated animals; however, there was a significant increase in hsp70 expression, apart from significant alteration in bcl-2 and bax at both mRNA transcript and protein expression level in the immune organs of aspartame treated animals compared to controls. Hence, the results indicated that hsp70 levels increased in response to oxidative injury induced by aspartame metabolites; however, these metabolites did not induce apoptosis in the immune organs. Furthermore, detailed analyses are needed to elucidate the precise molecular mechanisms involved in these changes.

  4. Upregulation of Bax and Bcl-2 following prenatal cocaine exposure induces apoptosis in fetal rat brain

    OpenAIRE

    Xiao, DaLiao; Zhang, Lubo

    2008-01-01

    Cocaine abuse during pregnancy has been associated with numerous adverse perinatal outcomes. Aims: The present study was to determine whether prenatal cocaine exposure induced apoptosis and the possible role of Bcl-2 family genes in the programming cell death in fetal rat brain. Main methods: Pregnant rats were treated with cocaine subcutaneously (30 & 60 mg/kg/day) from day 15 to 21 of gestation. Then the fetal and maternal brains were isolated. Key findings: Cocaine produced a dose-dependen...

  5. Upregulation of Bax and Bcl-2 following prenatal cocaine exposure induces apoptosis in fetal rat brain

    OpenAIRE

    DaLiao Xiao, Lubo Zhang

    2008-01-01

    Cocaine abuse during pregnancy has been associated with numerous adverse perinatal outcomes. Aims: The present study was to determine whether prenatal cocaine exposure induced apoptosis and the possible role of Bcl-2 family genes in the programming cell death in fetal rat brain. Main methods: Pregnant rats were treated with cocaine subcutaneously (30 & 60 mg/kg/day) from day 15 to 21 of gestation. Then the fetal and maternal brains were isolated. Key findings: Cocaine produced a dose-depe...

  6. Ganoderma lucidum spore powder modulates Bcl-2 and Bax expression in the hippocampus and cerebral cortex, and improves learning and memory in pentylenetetrazole-kindled rats

    Institute of Scientific and Technical Information of China (English)

    Shuang Zhao; Shengchang Zhang; Shuqiu Wang

    2011-01-01

    We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an epileptic group (kindled) and three medication groups ( 150, 300,450 mg/kg given to kindled rats). Bax and Bcl-2 immunohistochemistry and TUNEL labeling show ed that the number of Bax- and TUNEL-positive cells in the hippocampus and cerebral cortex decreased significantly in the high-dose medication group, while the number of Bcl-2immunoreactive cells increased. The Morris water maze test showed that high-dose treatment significantly shortened escape latency and increased spatial probe trial performance. Our findings indicate that a high dose of Ganoderma lucidum spore powder upregulates the expressionof antiapoptotic Bcl-2 protein in the hippocampus and cerebral cortex, inhibits proapoptotic Bax expression, and decreases seizure-induced neuronal apoptosis. Further,Ganoderma lucidum appears to protect against epilepsy-related learning and memory impairments.

  7. BaxSr1−xTi1.02O3 metal–insulator–metal capacitors on planarized alumina substrates

    NARCIS (Netherlands)

    Tiggelman, M.P.J.; Reimann, K.; Klee, M.; Mauczok, R.; Keur, W.; Hueting, R.J.E.

    2010-01-01

    Nanocrystalline barium strontium titanate (BaxSr1−xTi1.02O3) thin films with a barium content of x=0.8, 0.9 and 1 have been fabricated in a metal–insulator–metal configuration on glass-planarized alumina substrates. Cost-effective processing measures have been utilized by using poly-crystalline alum

  8. The trade-off between tuning ratio and quality factor of BaxSr1-xTiO3 MIM capacitors on alumina substrates

    NARCIS (Netherlands)

    Tiggelman, M.P.J.; Reimann, K.; Liu, J.; Klee, M.; Mauczok, R.; Keur, W.; Schmitz, J.; Hueting, R.J.E.

    2008-01-01

    Barium strontium titanate with different compositions is deposited using wet-chemical processing on a glass planarization layer, on top of alumina substrates. Three samples were fabricated with BaxSr1-xTiO3 (BST) with the barium content x varying between 0.8 and 1. The poly-crystalline films are 530

  9. Effect of compound preparation Tongqiao Jiannao capsules on neural cell apoptosis and Bcl-2 and Bax protein levels in a rat model of brain ischemia/reperfusion injury

    Institute of Scientific and Technical Information of China (English)

    Rui Wang; Guanglai Li; Wei Wang; Huanying Li

    2008-01-01

    BACKGROUND: Pharmacological studies have demonstrated that compound preparation Tongqiao Jiannao capsules composed of Zexie, Baizhu, Honghua, Danshen, and Shexiang can supplement qi,activate blood circulation, relieve blood stasis, induce resuscitation for alleviating pain, relieve pain, anddilate blood vessels.OBJECTIVE: To observe the effects of Tongqiao Jiannao capsules on the levels of the anti-apoptotic protein Bcl-2 and the pro-apoptotic protein Bax, and verify the mechanism of action.DESIGN, TIME AND SETTING: Randomized, controlled animal experiment, performed in the Laboratory of Biochemistry and Molecular Biology, Shanxi Medical University between June 2001 and December 2002.MATERIALS: The right middle cerebral arteries of 24 healthy adult Sprague Dawley rats were occluded by the suture method. The primary Chinese herbal medicinal ingredients of Tongqiao Jiannao capsules are Zexie. Baizhu, Honghua, Danshen, and Shexiang, which were purchased from Shanxi Provincial Medicinal Material Company, China, and prepared into condensed granules in the Room for Chinese Herbal Medicine Preparation, Second Hospital, Shanxi Medical University. Bcl-2 and Bax immunohistochemical staining kits, a 3,3-diaminobenzidine(DAB) kit, and an in situ apoptosis detection kit were purchased from Wuhan Boster Bioengineering Co., Ltd., China.METHODS: Twenty-four rats were randomly and evenly divided into three groups: (1) sham-operated rats in which sutures were inserted and immediately pulled out; (2) Tongqiao Jiannao capsule-treated rats that were intragastrically administered 6.5 g/kg/d Tongqiao Jiannao capsule preparation for seven successive days prior to middle cerebral artery occlusion (MCAO); and (3) MCAO rats without any other treatments.MAIN OUTCOME MEASURES: The levels of neural cell apoptosis and Bcl-2 and Bax proteins at 24 hours post-surgery.RESULTS: In the MCAO group, the numbers of apoptotic cells and Bax-positive cells were significantly increased, while the numbers of

  10. Runx3 Expression Inhibits Proliferation and Distinctly Alters mRNA Expression of Bax in AGS and A549 Cancer Cells

    Science.gov (United States)

    Torshabi, Maryam; Faramarzi, Mohammad Ali; Tabatabaei Yazdi, Mojtaba; Ostad, Seyyed Naser; Gharemani, Mohammad Hosein

    2011-01-01

    Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue–restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein. The GFP tagged Runx3 was transfected into AGS and A549 cells using fugene6 and PolyFect and Runx3 expression was confirmed by fluorescent microscopy and RT-PCR. The effect of Runx3 transfection on cell proliferation was determined by MTT assay and the results were confirmed by the trypan blue dye exclusion method. The effect of Runx3 expression on mRNA expression of BCL2-associated X protein (Bax) was evaluated using RT-PCR. In AGS and A549 cells, Runx3 expression inhibited cell proliferation (p < 0.01). The growth inhibition was less in A549 cells. We show that Runx3 expression increases Bax mRNA expression in AGS cells when compared with control (p < 0.05), but no significant differences in mRNA expression was observed in both examined cells. Runx3 expression has antiproliferative effect in AGS cell perhaps via increase in expression of Bax. The effect of Runx3 on A549 cells’ viability which has endogenous level of Runx3 is not related to Bax. These findings implicate a complex regulation by Runx3 in inhibition of cell proliferation utilizing Bax. PMID:24250365

  11. Study on Apoptosis and Expression of P53, Bcl-2, Bax in Cardiac Myocytys of Congestive Heart Failure Induced by Ventricular Pacing

    Institute of Scientific and Technical Information of China (English)

    QI; Benling; CAO; Linsheng; WANG; Lin; ZHOU; Jingqun

    2001-01-01

    The apoptosis and the expression of p53, bcl-2 and Bax in myocytes of chronic rapid ventricular pacing-induced congestive heart failure (CHF) in rabbits were investigated. The CHF rabbit model (P, n= 7) was established by chronic rapid ventricular pacing for 3 weeks. By using TUNEL technique the apoptosis in the myocytes in the rabbit model was studied and the expression of p53,bcl-2 and Bax in myocytes was detected by using immunohistochemical method. Sham-operated (C,n = 9) group served as control group. The results showed that there were about 4033± 884.56 apoptotic cells/106 myocytes in P group, but no apoptotic cells were found in C group. Myocytes positive for p53 immunoreactivity (18. 86±8. 48 vs 5. 06±0. 87, P<0.01) and positive for Bax immunoreactivity (7. 15±1.91 vs 0. 43±0. 09, P<0.01) were increased in P group as compared with those in C group, while the myocytes positive for bcl-2 immunoreactivity (7. 08±1.05 vs 14. 97±4.47,P<0. 01) and the ratio of bcl-2/Bax were decreased in P group as compared with those in C group.Apoptosis was involved in the development of CHF induced by continuously rapid ventricular pacing in rabbit. The expression of p53 and Bax was increased, while the expression of bcl-2 was inhibited.These might play an important role in the acceleration of the apoptosis.

  12. Photobiomodulation on Bax and Bcl-2 Proteins and SIRT1/PGC-1α Axis mRNA Expression Levels of Aging Rat Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    Fang-Hui Li

    2014-01-01

    Full Text Available Objective. This study aimed to analyze the effects of low level laser irradiation (LLLI on Bax and IGF-1 and Bcl-2 protein contents and SIRT1/PGC-1α axis mRNA expression levels to prevent sarcopenia in aged rats. Material and Methods. Twenty female Sprague Dawley rats (18 months old were randomly divided into two groups (n=10 per group: control (CON and LLLI groups. The gallium-aluminum-arsenium (GaAlAs laser irradiation at 810 nm was used in the single point contact mode (3.75 J/cm2; 0.4 cm2; 125 mW/cm2; 30 s. Bax, Bcl-2, and IGF-1 proteins and SIRT1/PGC-1α axis mRNA expression were assessed 24 h after LLLI on gastrocnemius in aged rat. Results. Gastrocnemius muscle weights, gastrocnemius mass/body mass, Bcl-2/BAX ratio, Bcl-2 protein, IGF-1 protein, and the mRNA contents in SIRT1, PGC-1α, NRF1, TMF, and SOD2 were significantly (P<0.05 increased by LLLI compared to CON group without LLLI. However, levels of BAX protein and caspase 3 mRNA were significantly attenuated by LLLI compared to CON group (P<0.05. Conclusion. LLLI at 810 nm inhibits sarcopenia associated with upregulation of Bcl-2/BAX ratio and IGF-1 and SIRT1/PGC-1α axis mRNA expression in aged rats. This indicates that LLLI has potential to decrease progression of myocyte apoptosis in sarcopenic muscles.

  13. Melatonin may play a role in modulation of bax and bcl-2 expression levels to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis

    Energy Technology Data Exchange (ETDEWEB)

    Mohseni, Mehran [Department of Radiology and Medical Physics, Faculty of Paramedicine, Kashan University of Medical Sciences, Kashan (Iran, Islamic Republic of); Mihandoost, Ehsan, E-mail: mihandoost.e@gmail.com [Department of Medical Radiation Engineering, Science and Research Branch, Islamic Azad University, Tehran (Iran, Islamic Republic of); Shirazi, Alireza [Department of Medical Physics and Biomedical Engineering, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Sepehrizadeh, Zargham [Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Bazzaz, Javad Tavakkoly [Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Ghazi-khansari, Mahmoud [Department of Pharmacology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2012-10-15

    The close relationship between free radicals effects and apoptosis process has been proved. Melatonin has been reported as a direct free radical scavenger. We investigated the capability of melatonin in the modification of radiation-induced apoptosis and apoptosis-associated upstream regulators expression in rat peripheral blood lymphocytes. Rats were irradiated with a single whole body Cobalt 60-gamma radiation dose of 8 Gy at a dose rate of 101 cGy/min with or without melatonin pretreatments at different concentrations of 10 and 100 mg/kg body weight. The rats were divided into eight groups of control, irradiation-only, vehicle-only, vehicle plus irradiation, 10 mg/kg melatonin alone, 10 mg/kg melatonin plus irradiation, 100 mg/kg melatonin alone and 100 mg/kg melatonin plus irradiation. Rats were given an intraperitoneal (IP) injection of melatonin or the same volume of vehicle alone 1 h prior to irradiation. Blood samples were taken 4, 24, 48 and 72 h after irradiation for evaluation of flow cytometric analysis of apoptotic lymphocytes using Annexin V/PI assay and measurement of bax and bcl-2 expression using quantitative real-time PCR (RT{sup 2}qPCR). Irradiation-only and vehicle plus irradiation showed an increase in the percentage of apoptotic lymphocytes significantly different from control group (P < 0.01), while melatonin pretreatments in a dose-dependent manner reduced it as compared with the irradiation-only and vehicle plus irradiation groups (P < 0.01) in all time points. This reduced apoptosis by melatonin was related to the downregulation of bax, upregulation of bcl-2, and therefore reduction of bax/bcl-2 ratio. Our results suggest that melatonin in these doses may provide modulation of bax and bcl-2 expression as well as bax/bcl-2 ratio to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis.

  14. Gene scanning and heart attack risk.

    Science.gov (United States)

    Barth, Andreas S; Tomaselli, Gordon F

    2016-04-01

    Coronary heart disease remains the leading cause of death in the Western World. The advent of microarray and next-generation sequencing technologies has generated multi-dimensional data sets, allowing for new pathophysiological insights into this complex disease. To date, genome-wide association studies (GWAS) have identified 152 associated loci and 320 candidate genes, contributing to the genetic risk of coronary artery disease (CAD) and acute myocardial infarction (AMI). The majority of single nucleotide polymorphisms (SNPs) mediate their risk by unknown mechanisms. A functional analysis based on Gene Ontology and KEGG pathways of candidate genes that are associated with CAD/AMI-SNPs showed the strongest evidence for genes regulating cholesterol metabolism. Additional clusters were significantly enriched for pathways, which play prominent roles during AMI and the development of atherosclerotic plaques in vascular tissue, including focal adhesion/extracellular matrix interaction, TGF-β signaling, apoptosis, regulation of vascular smooth muscle contraction, angiogenesis, calcium ion binding, and transcription factors. A systems genetics approach, which incorporates genetic risk with gene expression data, metabolomic data, and protein biochemistry into genome-wide network studies, holds promise to elucidate the complex interplay between genetic risk and environmental factors for coronary artery disease.

  15. BaxSr1−xTiO3 thin films prepared by OTS monomolecular film reverse induction and liquid phase self-assembly method

    International Nuclear Information System (INIS)

    Highlights: • OTS-SAMs were prepared on the substrate by self-assembled monomolecular technique. • After UV-light irradiation, OTS-SAMs became hydrophilic monolayers in 1 nm thickness. • Ferroelectric BaxSr1−xTiO3 film can be prepared on substrate by the reverse LPD-SAMs. -- Abstract: Octadecyl-trichlorosilane self-assembled monolayers (OTS-SAMs) were prepared on the ITO glass substrate surface by self-assembled monomolecular film technique and BaxSr1−xTiO3 thin film was prepared by the reverse induction and adsorption on the functionalized substrate surface. The morphologies of OTS-SAMs before and after UV-light irradiation, the variation of the contact angles, and the microstructure and electrical property of BaxSr1−xTiO3 thin film were investigated. The results show that after UV-light irradiation, the hydrophilic film of OTS-SAMs with the thickness of 1 nm and the contact angle of 5° can be used to prepare the homogeneous BaxSr1−xTiO3 thin film by the reverse induction method. As the increase of Ba content, the diffraction peaks corresponding to SrTiO3 crystal plane is shifted to small angles. The grains of the thin film are decreased as the increase of Ba doping content. When Sr/Ba is between 9/1 and 8/2, the dielectric constant of BaxSr1−xTiO3 thin film is higher while the dielectric loss is smaller. At 10 kHz, the dielectric constant of Ba0.2Sr0.8TiO3 thin film is 880 and the dielectric loss is 0.04. The remnant polarization of BaxSr1−xTiO3 thin film prepared by the reverse induction and adsorption and liquid phase self-assembly method is 0.64 μC/cm2 and the coercivity is 13.84 kV/cm

  16. Cyclophilin A affects Bcl-2 and Bax expression following beta-amyloid fragment 25-35-induced injury to PC12 cells

    Institute of Scientific and Technical Information of China (English)

    Li Cheng; Chaodong Zhang

    2008-01-01

    BACKGROUND: Cyclophilin A can protect neurons against oxidative stress.OBJECTIVE: To investigate the effect of cyclophilin A on Bcl-2 and Bax protein expression in pheochromocytoma (PCI2) cells treated with beta-amyloid fragment 25-35 (A β25-35), and to verify the protection pathway ofcyclophilin A.DESIGN, TIME AND SETTING: The initial experiment was performed at the Laboratory of Department of Neurology, First Clinical College, China Medical University from November 2006 to July 2007.MATERIALS: PCI2 cells were cultured at the Cell Center of Peking Union Medical College. A β25-35 (Sigma, USA), antibodies of Bcl-2 and Bax (Wuhan Boster, China), and recombinant human cyclophilin A (Biomol, USA) were used in this study.METHODS: PC12 cells were divided into three groups. Cells in the control group were incubated in culture medium. Cells in the Aβ25-35 injury group were incubated in medium containing a final concentration of 10 μ mol/L of Aβ25-35. Cells in the cyclophilin A group were incubated in medium containing a final concentration of 10 nmol/L of cyclophilin A for 30 minutes, and then treated with 10 μmol/L Aβ25-35. MAIN OUTCOME MEASURES: After 24 hours of culture, immunohistochemistry was used to detect Bcl-2 and Bax expression in PC12 cells. Annexin-V flow cytometry was employed to measure the apoptosis rate of PC12 cells. The MTT method was applied to examine the survival rate of PC12 cells.RESULTS: Bcl-2 expression decreased, whereas Bax expression increased in PCI2 cells treated with Aβ25-35 (t = 2.277, 5.957, P<0.05). However, in PC12 cells treated with Aβ25-35 and cyclophilin A, Bcl-2 expression increased and Bax expression decreased (t = 4.497, 2.531, P < 0.05). The survival rate of PC12 cells significantly decreased and the apoptosis rate increased (t=8.509, 22.886, P < 0.05) following Aβ25-35 treatment. Cyclophilin A enhanced the survival rate of PC12 cells to Aβ25-35-induced apoptosis (t = 4.895, 10.042, P< 0.05).CONCLUSION: Cyclophilin A can

  17. EXPRESSIONS OF survivin, bax AND caspase-3 IN MISSED ABORTION PATIENTS%稽留流产病人survivin、bax及caspase-3的表达

    Institute of Scientific and Technical Information of China (English)

    张娟; 王蓁

    2011-01-01

    目的 探讨survivin、bax及caspase-3的表达与稽留流产的关系.方法 以35例稽留流产病人为病例组,30例正常妊娠者为对照组.采用免疫组织化学sP法检测绒毛细胞滋养细胞和合体滋养细胞及蜕膜组织蜕膜细胞中survivin、bax和caspase-3的表达.结果 病例组妊娠组织中survivin的表达明显低于对照组,bax及caspase-3的表达明显高于对照组(u=2.87~5.29,P<0.01).病例组survivin与bax及caspase-3的表达均呈负相关(r=-0.59、-0.71,P<0.05、0.01),bax与caspase-3的表达呈正相关(r=0.79,P<0.01).结论 survivin的低表达、bax及caspase-3的高表达在稽留流产的发生中起重要作用.%Objective To study the relationship of expressions of survivin, bax and caspase-3 with missed abortion.Methods Thirty-five missed abortion patients were assigned as study group, and 30 with normal pregnancy as controls. Immunohistochemistry technique was employed to detect the expressions of survivin, bax and caspase-3 in cytotrophoblast and syncytiotrophoblast of villi and decidual cells of decidua. Results Compared with that of the controls, the expression of survivin in the study group was significantly lower, while the expressions of bax and caspase-3 were higher (uc = 2. 87-5.29, P<0.01 ). In the study group, the survivin expression was negatively correlated wit h t he expressions of bax and caspase-3 (r = - 0. 59,0. 71; P< 0. 05,0.01), respectively, and the expression of bax was found to be positively correlated with caspase-3 (r=0. 79,P<0.01). Conclusion Low expression of survivin and high expressions of bax and caspase-3 play an important role in the occurrence of missed abortion.

  18. Expression of bcl-2, bax in renal proximal tubular epithelial cells of rats with arsenic poisoning%bcl-2、bax在砷中毒大鼠肾近端小管表达

    Institute of Scientific and Technical Information of China (English)

    李远慧; 金婷婷

    2011-01-01

    Objective To investigate the influence of arsenic poisoning on the expressions of bcl-2, bax apoptosis control gene in renal proximal tubular epithelial cells in rtas.Methods Forty normal SD rats were divided into high and low dose of arsenic poisoning group and control group.The body weights of the rats were 120-150g.There were 15 rats in high and low dose exposure groups,and 10 rats in the control group.The rats in high and low groups were treated with As2O3 through drinking water at the doses of 10 and 0.4 mg/kg·d.The control group was given distilled water.Four months after the treatment,the kidney tissue of the rats was collected.Two step immunohistochemistry method, cell number count, and image analyses were used in the study.Results The bcl-2 immunoractive cells decreased and the average gray value gradually increased in arsenic poisoning groups(P < 0.05).The bax immunoractive cells of renal proximal tubular epithelial were increased and the average gray value decreased ( P < 0.05 ) in arsenic poisoning groups compared to those of the control group.Conclusion The expression of bcl-2, bax apoptosis control gene are involved in the process of apoptosis of renal proximal tubular epithelial cells in arsenic poisoning rats.%目的 探讨砷中毒对大鼠肾近端小管上皮细胞凋亡调控基因bcl-2、bax影响.方法 清洁级SD大鼠40只,体重为120~150g,高、低剂量染砷组各15只,对照组10只.高、低剂量染砷组分别给予三氧化二砷(AS2O3)10、0.4 mg/kg水溶液自由饮用,对照组饮用蒸馏水.分笼喂养4个月,取肾脏标本,采用免疫组织化学二步法、细胞计数和图像分析方法测定bcl-2、bax表达.结果 高、低剂量染砷组肾近端小管上皮bcl-2阳性细胞计数分别为(1.85±1.22)与(5.47±1.62)个,明显低于对照组(8.03±2.42)个,平均灰度值逐渐增高,差异具有统计学意义(P<0.05);高、低剂量染砷组肾近端小管上皮bax阳性细胞数分别为(14.88±3.02)与(6

  19. Role of the lattice dynamics in La2-xBaxCuO4 superconductor based on DFT method

    Directory of Open Access Journals (Sweden)

    A Tavana

    2010-09-01

    Full Text Available Electron-phonon coupling parameters are calculated for La2-x BaxCuO4 cuprate superconductor in a wide range of dopings, from undoped to overdoped compounds. In this study we aim to study the quality of such calculations based on DFT method so, the results of σ GGA+U electronic structure calculations are also investigated. The obtained value for electron-phonon coupling is in the same order of previous calculations but, the value obtained for the Hubbard U parameter shows that, such methods are poor in the estimation of electronic correlations to decide about the role of phonons in these compounds based on their results. Moreover, existence of several structural phase transitions with temperature and doping, lead to larger error in these calculations. Based on the calculated phonon dispersions, structural phase transitions can be resulted which shows the ability of DFT in the study of structural properties and the weakness of the strongly correlations in this properties.

  20. Grape Seed Procyanidin Extract Improves Insulin Production but Enhances Bax Protein Expression in Cafeteria-Treated Male Rats

    Directory of Open Access Journals (Sweden)

    Lídia Cedó

    2013-01-01

    Full Text Available In a previous study, the administration of a grape seed procyanidin extract (GSPE in female Wistar rats improved insulin resistance, reduced insulin production, and modulated apoptosis biomarkers in the pancreas. Considering that pharmacokinetic and pharmacodynamic parameters in females are different from these parameters in males, the aim of the present study was to evaluate the effects of GSPE on male Wistar cafeteria-induced obese rats. The results have confirmed that the cafeteria model is a robust model mimicking a prediabetic state, as these rats display insulin resistance, increased insulin synthesis and secretion, and increased apoptosis in the pancreas. In addition, GSPE treatment (25 mg/kg of GSPE for 21 days in male rats improves insulin resistance and counteracts the cafeteria-induced effects on insulin synthesis. However, the administration of the extract enhances the cafeteria-induced increase in Bax protein levels, suggesting increased apoptosis. This result contradicts previous results from cafeteria-fed female rats, in which GSPE seemed to counteract the increased apoptosis induced by the cafeteria diet.

  1. Spectroscopic studies of the ferroelectric and magnetic phase transitions in multiferroic Sr1-xBaxMnO3.

    Science.gov (United States)

    Goian, V; Kadlec, F; Kadlec, C; Dabrowski, B; Kolesnik, S; Chmaissem, O; Nuzhnyy, D; Kempa, M; Bovtun, V; Savinov, M; Hejtmánek, J; Prokleška, J; Kamba, S

    2016-05-01

    Dielectric response of perovskite Sr1-xBaxMnO3 (x = 0.43 and 0.45) ceramics was investigated using microwave, THz and infrared spectroscopic techniques in order to study the ferroelectric and antiferromagnetic phase transitions with critical temperatures TC ≈ 350 K and TN ≈ 200 K, respectively. The two lowest-frequency polar phonons are overdamped above TN and they exhibit pronounced softening on heating towards TC. Nevertheless, permittivity ε' in the THz range shows only a small anomaly at TC because the phonon contribution to ε' is rather small. The phonons are coupled with a central mode which provides the main contribution to the dielectric anomaly at TC. Thus, the ferroelectric phase transition has characteristics of a crossover from displacive to order-disorder type. At the same time, the intrinsic THz central peak is partially screened by conductivity and related Maxwell-Wagner relaxation, which dominates the microwave and lower-frequency spectra. Below TN, the ferroelectric distortion markedly decreases, which has an influence on the frequencies of both the central and soft modes. Therefore, ε' in the THz range increases at TN on cooling. In spite of the strong spin-phonon coupling near TN, surprisingly no magnetodielectric effect was observed in the THz spectra upon applying magnetic field of up to 7 T, which is in contradiction with the theoretically expected huge magnetoelectric coupling. We explain this fact as due to the insensitivity of TN to magnetic field. PMID:27023160

  2. The N-terminus and alpha-5, alpha-6 helices of the pro-apoptotic protein Bax, modulate functional interactions with the anti-apoptotic protein Bcl-xL

    Directory of Open Access Journals (Sweden)

    Sowdhamini R

    2007-05-01

    Full Text Available Abstract Background Bcl-2 family proteins are key regulators of mitochondrial integrity and comprise both pro- and anti-apoptotic proteins. Bax a pro-apoptotic member localizes as monomers in the cytosol of healthy cells and accumulates as oligomers in mitochondria of apoptotic cells. The Bcl-2 homology-3 (BH3 domain regulates interactions within the family, but regions other than BH3 are also critical for Bax function. Thus, the N-terminus has been variously implicated in targeting to mitochondria, interactions with BH3-only proteins as well as conformational changes linked to Bax activation. The transmembrane (TM domains (α5-α6 helices in the core and α9 helix in the C-terminus in Bax are implicated in localization to mitochondria and triggering cytotoxicity. Here we have investigated N-terminus modulation of TM function in the context of regulation by the anti-apoptotic protein Bcl-xL. Results Deletion of 29 amino acids in the Bax N-terminus (Bax 30–192 caused constitutive accumulation at mitochondria and triggered high levels of cytotoxicity, not inhibited by Bcl-xL. Removal of the TM domains (Bax 30–105 abrogated mitochondrial localization but resulted in Bcl-xL regulated activation of endogenous Bax and Bax-Bak dependent apoptosis. Inclusion of the α5-α6 helices/TMI domain (Bax 30–146 phenocopied Bax 30–192 as it restored mitochondrial localization, Bcl-xL independent cytotoxicity and was not dependent on endogenous Bax-Bak. Inhibition of function and localization by Bcl-xL was restored in Bax 1–146, which included the TM1 domain. Regardless of regulation by Bcl-xL, all N-terminal deleted constructs immunoprecipitated Bcl-xLand converged on caspase-9 dependent apoptosis consistent with mitochondrial involvement in the apoptotic cascade. Sub-optimal sequence alignments of Bax and Bcl-xL indicated a sequence similarity between the α5–α6 helices of Bax and Bcl-xL. Alanine substitutions of three residues (T14A-S15A-S16A in

  3. Ab-initio study of structural, elastic, electronic and thermodynamic properties of BaxSr1−xS ternary alloys

    Directory of Open Access Journals (Sweden)

    Chelli S.

    2015-12-01

    Full Text Available The structural, elastic, electronic and thermodynamic properties of BaxSr1−xS ternary alloys have been investigated using the full-potential (linearized augmented plane wave method. The ground state properties, such as lattice constant, bulk modulus and elastic constants, are in good agreement with numerous experimental and theoretical data. The dependence of the lattice parameters, bulk modulus and band gap on the composition x was analyzed. Deviation of the lattice constant from Vegard’s law and the bulk modulus from linear concentration dependence (LCD was observed. The microscopic origins of the gap bowing were explained by using the approach of Zunger et al. The thermodynamic stability of BaxSr1−xS alloy was investigated by calculating the excess enthalpy of mixing, ΔHm and the calculated phase diagram showed a broad miscibility gap with a critical temperature.

  4. Injury of mitochondria and the expressions of fas and bax mRNA in the hip-pus of epileptic rats of different latency

    Institute of Scientific and Technical Information of China (English)

    Shuhai Tang; Jianying Sun; Xiaojun Pan; Li Zhang

    2006-01-01

    BACKGROUND: It has been confirmed that Fas and Bax respectively mediate the exogenous and endogenous pathways of neuronal apoptosis, and then mediate the neuronal injury after status epilepticus.OBJECTIVE: To comparatively observe the injury of mitochondrial ultrastructure and the expressions of fas and bax in hippocampal tissue of rats with status epilepticus of different latency.DESIGN: A randomized control study.SETTING: Department of Anesthesiology and Department of Neurology, Qilu Hospital of Shandong University.MATERIALS: Totally 110 male adult SD rats of 260-300 g were used. Kainic acid was purchased from American Sigma Company.METHODS: The experiment was carried out in the Pathological Laboratory of Shandong Academy of Medical Sciences between March and July 2005.① Totally 100 SD rats were divided into two groups according to the random number table method:intraperitoneal injection group and caudal venous injection group.The rats were given kainic acid injected intraperitoneally(12 mg/kg)and through caudal vein (10 mg/kg) respectively. Each group was observed at 3, 6, 24, 48 and 72 hours after status epilepticus respectively.Ten rats were selected for each time point, including 2 for examination of electron microscope and 8 for the diction of the fas and bax mRNA expressions. The time and manifestations of seizure were observed, and the seizure was lasted for 2 hours, and then it was terminated by intraperitoneal injection of diazepam (10 mg/kg). Another 10 rats were used as the normal control group, and the materials were taken at 24 hours after status epilepticus, 2 of rats for the examination of electron microscope and 8 of them for the reverse transcription-polymerase chain reaction (RT-PCR). ② The ultrastructure of hippocampal neurons and its mitochondria were observed with transmission electron microscope. ③ The fas and bax mRNA expressions were detected with reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: The

  5. Titanium Dioxide (TiO2) Nanoparticles Preferentially Induce Cell Death in Transformed Cells in a Bak/Bax-Independent Fashion

    OpenAIRE

    Yanglong Zhu; Eaton, John W.; Chi Li

    2012-01-01

    While the cytotoxic effects of titanium dioxide (TiO(2)) nanoparticles have been under intense investigation, the molecular mechanisms of this cytotoxicity remain unknown. Here we investigated the influence of oncogenic transformation and a major apoptotic signaling pathway on cellular responses to TiO(2) nanoparticles. Isogenic wild-type (WT) and apoptosis-resistant (Bak(-/-)Bax(-/-)) cell lines with and without tumorigenic transformation were examined. TiO(2) nanoparticles preferentially re...

  6. Effect of Achyranthes Bidentata Polysaccharides on the Expression of BCL-2 and Bax in Hepatic Tissues after Exhaustive Exercise in Rats

    OpenAIRE

    Lin, Jinyang; Zhang, Zhuoying; Shan, Ying

    2010-01-01

    This study aims to assess the effects of Achyranthes bidentata polysaccharides (ABPS) on the expression of bcl-2 and bax in hepatic tissues after exhaustive exercise in order to provide theoretical support for the application of ABPS in the field of sports nutrition. Thirty male Sprague-Dawley rats were randomized into three groups, each consisting of 10 rats: Normal control group (NCG), Exhausting exercises control group (EECG), ABPS treated group (ATG). ABPS were fed orally by gastric intub...

  7. An Assay of Bax and Bcl2 Expression in Mice Hippocampus Following Ischemia-Reperfusion Treatment with CoQ10

    Directory of Open Access Journals (Sweden)

    Jalal Hassanshahi

    2013-10-01

    Full Text Available Introduction: Preliminary studies confirmed reduction of cell death following treatment with antioxidants. According to this finding we investigated the relationship between consumption of CoQ10 and expression of bax and bcl2 in hippocampus ischemia that this expression related to cell programmed death.Material and Methods: We studied the protective role of CoQ10 against ischemia-reperfusion. Experimental design includes four groups: intact (N=7, ischemic control (N=7, sham control (N=7 and treatment groups with CoQ10 (N=7. The mice (treatment group treated with CoQ10 as Pre-Treatment for a week. Then, ischemia induced by common carotid artery ligation and following the reduction in inflammation (a week the treatment group post-treated with CoQ10 for a week. Nissl staining applied to counting necrotic cells of hippocampus and the western blotting performed to measurement the bax and bcl2 expression. Tunnel kit was used to quantify apoptotic cell death while to short term memory scale, we apply Y-maze.Results: Cell death was significantly lower when mice treated with CoQ10. Bax expression was significantly high in ischemic group but in treatment group was less and reversely the bcl2 expression in ischemic group was lower than treatment and vehicle groups. The memory test results were consistent with cell death results. Conclusion: Ischemia for 15 minutes induced cell death in hippocampus with more potent effect on CA1. CoQ10 intake significantly reduced cell death and decreased memory loss. with prevent of expression of bax and increase in expression of bcl2.

  8. Angiotensin II-Induced Apoptosis of Human Umbilical Vein Endothelial Cells was Inhibited by Blueberry Anthocyanin Through Bax- and Caspase 3-Dependent Pathways.

    Science.gov (United States)

    Du, Jian; Leng, Jiyan; Zhang, Li; Bai, Guangxin; Yang, Di; Lin, Huan; Qin, Junjie

    2016-01-01

    BACKGROUND This study aimed to investigate the inhibitory effect of blueberry anthocyanin (BBA) on Angiotensin II (Ang II)-induced apoptosis of human umbilical vein endothelial cells (HUVECs), and its regulation mechanisms involving Bax and Caspase 3. MATERIAL AND METHODS HUVECs were first treated by different concentrations of Ang II (10-9, 10-8, 10-7, 10-6, 10-5, and 10-4 mol/L) and BBA (80, 40, 20, 10, 5, and 2.5 μg/ml). After 24 h and 48 h of treatment, MTT was performed to detect the viability of HUVECs. Then, HUVECs were randomly divided into the Ang II group (10-6 mol/L Ang II) and Ang II + BBA group (10-6 mol/L Ang II and 20 μg/ml BBA), and the apoptosis rate was detected by flow cytometry. Western blot analysis was performed to detect the expression of Bax and Caspase 3 in these 2 groups. During the whole process, HUVECs without any treatments served as the control group. RESULTS The cell viability of HUVECs was significantly reduced by Ang II in a time- and concentration-dependent manner (P<0.05), while BBA significantly elevated the cell viability of HUVECs until a peak of 20.0 μg/ml. The apoptosis rate of HUVECs was significantly increased by Ang II (P<0.01) and reduced by the BBA intervention (P<0.05). Ang II significantly elevated the expression of Bax and Caspase 3 in HUVECs, but their expression was significantly inhibited by BBA. CONCLUSIONS BBA increased cell viability and reduced apoptosis rate of HUVECs induced by Ang II through Bax- and Caspase 3-dependent pathways. PMID:27616275

  9. A novel strategy for cancer gene therapy: RNAi

    Institute of Scientific and Technical Information of China (English)

    PAN Qiuwei; CAI Rong; LIU Xinyuan; QIAN Cheng

    2006-01-01

    RNA interference (RNAi) induces genesilencing at a level of posttranscription mediated bydouble stranded RNA. There are numerous methods for delivery of small double-stranded interference RNA (siRNA) to the target cells, including nonviral and viral vectors. Among these methods, viral vectors are the more efficient vehicles. The expression of short hairpin RNA (shRNA) by viral vectors in target cells can be cut by Dicer enzyme to become ~21 bp siRNA, which could guide degradation of cognate mRNA. RNAi technology can be directed against cancer using a variety of strategies, including the inhibition of overexpressed oncogenes, promoting apoptosis, regulating cell cycle, antiangiogenesis and enhancing the efficacy of chemotherapy and radiotherapy. Since RNAi technology has become an excellent strategy for cancer gene therapy, this review outlines the latest developments and applications of such a novel technology.

  10. Oridonin induces apoptosis of HeLa cells via altering expres sion of Bcl-2/Bax and activating caspase-3/ICAD pathway

    Institute of Scientific and Technical Information of China (English)

    Chun-ling ZHANG; Li-jun WU; Shin-ichi TASHIRO; Satoshi ONODERA; Takashi IKEJIMA

    2004-01-01

    AIM: To study the mechanisms by which oridonin inhibited HeLa cell growth in vitro. METHODS: Viability of oridonin-induced HeLa cells was measured by MTT assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis.Caspase activity was assayed using fiuorometric protease assay. ICAD, Bcl-2, and Bax proteins expression were detected by Western blot analysis. RESULTS: Oridonin induced oligonucleosomal fragmentation of DNA and increased caspase-3 activity, on the other hand, reduced the expression of inhibitor of caspase-3-activated DNase (ICAD), a caspase-3 substrate, at 12 h in HeLa cells. Oridonin-induced DNA fragmentation, caspase-3 activation and down-regulation of ICAD expression were effectively inhibited by a caspase-3 inhibitor, z-DEVD-fmk (z-AspGlu-Val-Asp-fmk). However, pretreatment with an inhibitor of poly (ADP-ribose) polymerase (PARP), 3, 4-dihydro5-[4-(1-piperidinyl)butoxy]-1 (2H)-isoquinolinone (DPQ), did not suppress oridonin-induced HeLa cell death. In addition, oridonin-induced apoptosis was associated with an increase in the expression of the apoptosis inducer Bax, and a significant reduction in expression of the apoptosis suppressor Bcl-2 in mitochondria. CONCLUSION:Oridonin induces HeLa cells apoptosis by altering balance of Bcl-2 and Bax protein expression and activation of caspase-3/ICAD pathway.

  11. Gambogic acid induces mitochondria-dependent apoptosis by modulation of Bcl-2 and Bax in mantle cell lymphoma JeKo-1 cells

    Institute of Scientific and Technical Information of China (English)

    Jingyan Xu; Min Zhou; Jian Ouyang; Jing Wang; Qiguo Zhang; Yong Xu; Yueyi Xu

    2013-01-01

    Objective:To study the mechanisms in gambogic acid (GA)-induced JeKo-1 human Mantle Cell Lymphoma cell apoptosis in vitro.Methods:The proliferation of GA-treated JeKo-1 cells was measured by CCK-8 assay and Ki-67 immunocytochemical detection.Apoptosis,cell cycle and mitochondrial membrane potential were measured by flow cytometric analysis.Caspase-3,-8 and-9 were detected by colorimetric assay.Bcl-2 and Bax were analyzed by Western blotting.Results:GA inhibited cell growth in a time-and dose-dependent manner.GA induces apoptosis in JeKo-1 cells but not in normal bone marrow cells,which was involved in reducing the membrane potential of mitochondria,activating caspases-3,-8 and-9 and decreasing the ratio of Bcl-2 and Bax without cell cycle arresting.Conclusions:GA induced apoptosis in human MCL JeKo-1 cells by regulating Bcl-2/Bax and activating caspase-3,-8 and-9 via mitochondrial pathway without affecting cell cycle.

  12. Effect of Hypoxic Preconditioning on Neural Cell Apoptosis and Expression of Bcl-2 and Bax in Cerebral Ischemia-Reperfusion in Rats

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    In order to investigate the protective effect of hypoxic preconditioning on the cerebral ischemia-reperfusion injury, the expression of Bcl-2 and Bax was detected by using immunohistochemical staining after 3 h cerebral ischemia followed by 1, 6, 12, 24 and 48 h reperfusion respectively in rats treated with or without hypoxic preconditioning before cerebral ischemia. In addition,the apoptosis of neural cells and the behavioral scores for neurological functions recovery were evaluated by TUNEL staining and "crawvling method", respectively. Compared with control group (cerebral ischemia-reperfusion without hypoxic preconditioning), the expression of Bcl-2 was significantly increased, but that of Bax decreased in the hypoxic preconditioning group (cerebral ischemiareperfusion with hypoxic preconditioning), both P<0. 05. The pre-treatment with hypoxic preconditioning could reduce the apoptosis of neural cells and promote the neurological function recovery as compared to control group. It was suggested that hypoxic preconditioning may have protective effects on the cerebral ischemia-reperfusion injury by inhibiting the apoptosis of neural cells, increase the expression of Bcl-2 and decrease the expression of Bax.

  13. Gastroprotective activity of Annona muricata leaves against ethanol-induced gastric injury in rats via Hsp70/Bax involvement.

    Science.gov (United States)

    Moghadamtousi, Soheil Zorofchian; Rouhollahi, Elham; Karimian, Hamed; Fadaeinasab, Mehran; Abdulla, Mahmood Ameen; Kadir, Habsah Abdul

    2014-01-01

    The popular fruit tree of Annona muricata L. (Annonaceae), known as soursop and graviola, is a widely distributed plant in Central and South America and tropical countries. Leaves of A. muricata have been reported to possess antioxidant and anti-inflammatory activities. In this study, the gastroprotective effects of ethyl acetate extract of A. muricata leaves (EEAM) were investigated against ethanol-induced gastric injury models in rats. The acute toxicity test of EEAM in rats, carried out in two doses of 1 g/kg and 2 g/kg, showed the safety of this plant, even at the highest dose of 2 g/kg. The antiulcer study in rats (five groups, n=6) was performed with two doses of EEAM (200 mg/kg and 400 mg/kg) and with omeprazole (20 mg/kg), as a standard antiulcer drug. Gross and histological features showed the antiulcerogenic characterizations of EEAM. There was significant suppression on the ulcer lesion index of rats pretreated with EEAM, which was comparable to the omeprazole effect in the omeprazole control group. Oral administration of EEAM to rats caused a significant increase in the level of nitric oxide and antioxidant activities, including catalase, glutathione, and superoxide dismutase associated with attenuation in gastric acidity, and compensatory effect on the loss of gastric wall mucus. In addition, pretreatment of rats with EEAM caused significant reduction in the level of malondialdehyde, as a marker for oxidative stress, associated with an increase in prostaglandin E2 activity. Immunohistochemical staining also demonstrated that EEAM induced the downregulation of Bax and upregulation of Hsp70 proteins after pretreatment. Collectively, the present results suggest that EEAM has a promising antiulcer potential, which could be attributed to its suppressive effect against oxidative damage and preservative effect toward gastric wall mucus.

  14. Effects of in vitro fertilization and embryo culture on TRP53 and Bax expression in B6 mouse embryos

    Directory of Open Access Journals (Sweden)

    Chami Omar

    2006-11-01

    Full Text Available Abstract In the mouse, embryo culture results in a characteristic phenotype of retarded embryo preimplantation development and reduced numbers of cells within embryos. The expression of TRP53 is central to the regulation of the cell's capacity to proliferate and survive. In this study we found that Trp53 mRNA is expressed throughout the preimplantation stage of development. Levels of TRP53 protein expression were low during the cleavage stages and increased at the morula and blastocyst stages in B6 embryos collected from the reproductive tract. Embryos collected at the zygote stage and cultured for 96 h also showed low levels of TRP53 expression at precompaction stages. There were higher levels of TRP53 in cultured morula and the level in cultured blastocysts was clearly increased above blastocysts collected directly from the uterus. Immunolocalization of TRP53 showed that its increased expression in cultured blastocysts corresponded with a marked accumulation of TRP53 within the nuclei of embryonic cells. This pattern of expression was enhanced in embryos produced by in vitro fertilization and subjected to culture. The TRP53 was transcriptionally active since culture also induced increased expression of Bax, yet this did not occur in embryos lacking Trp53 (Trp53-/-. The rate of development of Trp53-/- zygotes to the blastocyst stage was not different to wildtype controls when embryos were cultured in groups of ten but was significantly faster when cultured individually. The results show that zygote culture resulted in the accumulation of transcription activity of TRP53 in the resulting blastocysts. This accounts for the adverse effects of culture of embryos individually, but does not appear to be the sole cause of the retarded preimplantation stage growth phenotype associated with culture in vitro.

  15. Transcription of five p53- and Stat-3-Inducible genes after ionizing radiation

    International Nuclear Information System (INIS)

    Ionizing radiation (IR) produces temporal- and dose-dependent changes in multiple gene mRNA targets that are potential biomarkers of radiation dose. We confirmed IR-induced changes in expression of gadd45a, ddb-2, and cdkn1a downstream transcripts of p53 by quantitative reverse transcription-polymerase chain reaction (QRT-PCR) assay in total RNA samples from the whole blood of radiotherapy patients undergoing total-body irradiation [Amundson, S.A., Grace, M.B., McLeland, C.B., Epperly, M.W., Yeager, A., Zhan, Q., Greenberger, J.S., Fornace Jr., A.J., 2004. Human in vivo radiation-induced biomarkers: gene expression changes in radiotherapy patients. Cancer Res. 64, 6368-6371.]. We now confirm dose-dependent up-regulation of bax in addition to these p53-dependent transcripts, and bcl-2, a downstream transcript of Stat-3, in ex vivo irradiated blood samples from healthy unrelated volunteers. Together these biomarkers represent pathways involved in growth arrest, DNA damage, and apoptosis. The objectives of this study were to (1) investigate the relationship between baseline mRNA expression levels, and (2) define expression patterns in response to IR in a large cohort (n=20). Whole-blood samples were irradiated ex vivo to measure gene expression in samples from (i) three healthy donors over a broad dose range (0, 0.25, 0.50, 0.75, 1, 2, and 3 Gy), and (ii) 20 healthy donors at two doses, 0.25 and 2.5 Gy. Expression level variance (σ2) of baseline values (0 Gy) showed negligible inter-individual variation with all values ≤1.0. σ2values=0.50bax, 0.25 bcl-2, 0.73 gadd45a, 0.66 cdkn1a, and 1.0 ddb-2. Meaningful IR dose-responses were observed for bax, gadd45a, and ddb-2 profiles and the ratio of bax:bcl-2 mRNA expression over a broad dose range. QRT-PCR studies were extended in the lower dose range (0, 0.1, 0.5, 0.75, and 1 Gy). Results showed that bax:bcl-2 ratio initially favors bax expression at doses of <1Gy, with IR-induced dose responses observed between 1 and 3

  16. Alternation of apoptotic and implanting genes expression of mouse embryos after re-vitrification

    Science.gov (United States)

    Majidi Gharenaz, Nasrin; Movahedin, Mansoureh; Mazaheri, Zohreh; Pour beiranvand, Shahram

    2016-01-01

    Background: Nowadays, oocytes and embryos vitrification has become a routine technique. Based on clinical judgment, re-vitrification maybe required. But little is known about re-vitrification impact on genes expression. Objective: The impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study. Materials and Methods: In this experimental study, 8 cell embryos (n=240) were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh (n=80), vitrified at 8 cell stage (n=80), vitrified at 8 cell stage thawed and re-vitrified at compaction stage (n=80). Embryos were vitrified by using cryolock, (open system) described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts. Results: Our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos (p=0.03). In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos (p=0.004), however expression of Bax and Bcl-2 (apoptotic) genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos (p=0.003), but it was similar between re-vitrified and vitrified embryos. Conclusion: Re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage. PMID:27679826

  17. Exercise Preconditioning Decreases the Ratio of bax/bcl-2 mRNA Expression in Midbrain of Mouse Model with Parkinson's Disease%运动预适应下调帕金森小鼠中脑bax/bcl-2 mRNA表达比值

    Institute of Scientific and Technical Information of China (English)

    薛宏斌; 张勇; 刘洪涛; 马强

    2009-01-01

    目的:探讨运动预适应是否下调帕金森小鼠中脑细胞bax/bcl-2 mRNA表达比值.方法:32只雄性小鼠(8~9周龄)首先随机分为安静组、运动组.运动组连续5周,每天1次中等强度(12米/分,20分钟)跑台训练.然后以上两组再各自随机分成两组,于训练结束后第2天分别注射生理盐水或中等剂量MPTP(30mg/kg×2次).只有表现典型行为变化的MPTP小鼠被认为是成功帕金森模型小鼠,进入最终分组,分为安静+生理盐水组(Se+Sa)、运动+生理盐水组(E+Sa)、安 +MPTP组(Se+M)、运 +MPTP组(E+M)4组,每组8只.训练结束后第11天处死动物,检测小鼠中脑bax、bcl-2 mRNA表达,计算bax/bcl-2 mRNA表达比值.结果:E+M组小鼠中脑bax mRNA表达较Se+M组显著降低(P<0.05),E+M组bcl-2 mRNA表达较Se+M组显著增加(P<0.05),E+M组bax/bcl-2比值显著小于Se+M组(P<0.05).结论:运动预适应改变帕金森小鼠中脑黑质细胞凋亡信号分子bcl-2、bax mRNA表达,降低bax/bcl-2 mRNA表达比值.

  18. Ginkgo biloba leaf extract effects on inducible nitric oxide synthase, Bcl-2, and Bax expression in rat models of spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Jiejun Jiao; Bin Du

    2008-01-01

    BACKGROUND: Ginkgo biloba leaf extract exhibits neuroprotective effects in spinal cord injury. However,the mechanisms of action remain unclear.OBJECTIVE: To investigate inducible nitric oxide synthase (iNOS) and Bcl-2/Bax expression in the injured spinal cord, and to explore the neuroprotective mechanisms of ginkgo biloba leaf extract in rats with spinal cord injury.DESIGN, TIME AND SETTING: The randomized, controlled, cell molecular biology experiment was performed at Soochow University, China from March 2007 to March 2008.MATERIALS: A total of 120 healthy, adult Sprague Dawley rats were selected for this study. Rat models of moderate acute thoracic (T9) spinal cord injury were established using the modified Allen method.Shuxuening injection was obtained from Zhenbaodao Pharmaceutical Co., Ltd., China. Methylprednisolone was purchased from North China Pharmaceutical Co., Ltd.METHODS: All rats were equally and randomly divided into four groups. Only the spinal cord was exposed in the sham operation group rats. In the trauma group, rats were not treated with drugs following spinal cord injury. Rats in the hormone group were intraperitoneally injected with 30 mg/kg methylprcdnisolone following spinal cord injury. Rats in the ginkgo biloba leaf extract group were intraperitoneally infused with a 1.0 mL/kg Shuxuening injection per day.MAIN OUTCOME MEASURES: At l hour, as well as 1, 3, 5, 7, and 14 days after spinal cord injury,iNOS- and Bcl-2/Bax-positive cells were quantified with immunohistochemistry. Pathological changes were detected using hematoxylin-eosin staining under an optical microscope.RESULTS: Spinal cord injury in the ginkgo biloba leaf extract and hormone groups was milder compared with the trauma group. Demyelination was significantly ameliorated and the necrotic cavity was obviously reduced in the injured spinal cord of rats in the ginkgo biloba leaf extract and hormone groups at each time point, iNOS expression was increased in the injured spinal cord

  19. Effects of exercises on mitochondrial permeability transition pore,and bcl-2 and bax mRNA expression in rat skeletal muscle%骨骼肌细胞线粒体通透性转换孔及bcl-2和bax mRNA表达与运动

    Institute of Scientific and Technical Information of China (English)

    王冬梅; 漆正堂; 丁树哲

    2011-01-01

    BACKGROUND:Exercises influence the apoptosis of skeletal muscle cells and mitochondrial pathway is one of important pathways mediating cellular apoptosis.OBJECTIVE:To investigate the effects of exercises on mitochondrial permeability transition pore, and bcl-2 and bax mRNA expression in rat skeletal muscle .METHODS:Twenty-four Sprague-Dawley male rats were randomly divided into three groups: the control group, 6-week swimming training group (six swimming trainings per week) and one -off exhaustive swimming group (one -off exhaustive swimming exercise).The opening state of mitochondrial permeability transition pore in skeletal muscle was examined with ultraviolet spectrophotometer,and the mRNA expression of bcl-2 and bax in rat skeletal muscle was determined by reverse transcription-polymerase chain reaction.RESULTS AND CONCLUSION:6-week swimming training resulted in significant increase of bcl-2 mRNA expression and significant decrease of bax mRNA expression , as well as significant increase of bcl-2lbax mRNA (P < 0.01) but the opening state of mitochondrial permeability transition pore was not altered obviously, compared with the control group. The opening state of mitochondrial permeability transition pore was significantly increased (P < 0.01), bcl-2 mRNA expression was significantly decreased, bax mRNA expression was significantly increased, bcl-2/bax mRNA expression was significantly decreased in the one-off exhaustive swimming group than in the control group (P < 0.01). These findings suggest that exercise training can regulate skeletal muscle cell apoptosis by altering the opening stage of mitochondrial permeability transition pore and regulating bcl-2/bax mRNA expression in rat skeletal muscle.%背景:运动影响骨骼肌细胞的凋亡,而线粒体途径是介导细胞凋亡的一个重要途径.目的:研究运动对大鼠骨骼肌线粒体通透性转换孔、凋亡调控基因bcl-2 和bax 表达的影响.方法:将24 只成

  20. Expression of epithelial-mesenchymal transition-related genes increases with copy number in multiple cancer types.

    Science.gov (United States)

    Zhao, Min; Liu, Yining; Qu, Hong

    2016-04-26

    Epithelial-mesenchymal transition (EMT) is a cellular process through which epithelial cells transform into mesenchymal cells. EMT-implicated genes initiate and promote cancer metastasis because mesenchymal cells have greater invasive and migration capacities than epithelial cells. In this pan-cancer analysis, we explored the relationship between gene expression changes and copy number variations (CNVs) for EMT-implicated genes. Based on curated 377 EMT-implicated genes from the literature, we identified 212 EMT-implicated genes associated with more frequent copy number gains (CNGs) than copy number losses (CNLs) using data from The Cancer Genome Atlas (TCGA). Then by correlating these CNV data with TCGA gene expression data, we identified 71 EMT-implicated genes with concordant CNGs and gene up-regulation in 20 or more tumor samples. Of those, 14 exhibited such concordance in over 110 tumor samples. These 14 genes were predominantly apoptosis regulators, which may implies that apoptosis is critical during EMT. Moreover, the 71 genes with concordant CNG and up-regulation were largely involved in cellular functions such as phosphorylation cascade signaling. This is the first observation of concordance between CNG and up-regulation of specific genes in hundreds of samples, which may indicate that somatic CNGs activate gene expression by increasing the gene dosage.

  1. Garlic ((Allium sativum)) Fresh Juice Induces Apoptosis in Human Oral Squamous Cell Carcinoma: The Involvement of Caspase-3, Bax and Bcl-2.

    Science.gov (United States)

    Farhadi, Farrokh; Jahanpour, Salar; Hazem, Kameliya; Aghbali, Amirala; Baradran, Behzad; Vahid Pakdel, Seyyed Mahdi

    2015-01-01

    Background and aims. There is no report on the apoptotic impact of Allium sativum L.(Garlic) on the oral squamous cell carcinoma (KB); hence, this study was designed to survey the apoptotic effects of garlic fresh juice (GFJ) on the KB cells. Materials and methods. MTTassay (MicrocultureTetrazolium Assay) was carried out to evaluate the cytotoxicity of GFJ on KB cells. Furthermore, TUNEL(Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling)and DNA fragmentation tests were performed to determine if GFJ is able to induce apoptosis in KB cells. Also a standard kit was used to assess caspase-3 activity in KB cells. Also western blotting was employed to evaluate the effect of GFJ on Bax:Bcl-2 ratio. Results. Significant cytotoxic effects were observed for the minimum used concentration (1μg/mL) as calculated to be 77.97±2.3% for 24 h and 818±3.1% for 36h of incubation (P < 0.001). Furthermore, TUNEL and DNA fragmentation tests corroborated the apoptosis inducing activity of GFJ. Consistently, after treating KB cells with GFJ(1μg/mL), caspase-3 activity and Bax:Bcl-2 ratio were raised by 7.3±0.6 and (P <0.001) folds, respectively. Conclusion. The results of this study advanced that GFJ induces apoptosis in the KB cells through increasing caspase-3 activity and Bax:Bcl2 ratio which could be attributed to its organo-sulfurcomponents. PMID:26889365

  2. Dexamethasone protected human glioblastoma U87MG cells from temozolomide induced apoptosis by maintaining Bax:Bcl-2 ratio and preventing proteolytic activities

    Directory of Open Access Journals (Sweden)

    Patel Sunil J

    2004-12-01

    Full Text Available Abstract Background Glioblastoma is the deadliest and most prevalent brain tumor. Dexamethasone (DXM is a commonly used steroid for treating glioblastoma patients for alleviation of vasogenic edema and pain prior to treatment with chemotherapeutic drugs. Temozolomide (TMZ, an alkylating agent, has recently been introduced in clinical trials for treating glioblastoma. Here, we evaluated the modulatory effect of DXM on TMZ induced apoptosis in human glioblastoma U87MG cells. Results Freshly grown cells were treated with different doses of DXM or TMZ for 6 h followed by incubation in a drug-free medium for 48 h. Wright staining and ApopTag assay showed no apoptosis in cells treated with 40 μM DXM but considerable amounts of apoptosis in cells treated with 100 μM TMZ. Apoptosis in TMZ treated cells was associated with an increase in intracellular free [Ca2+], as determined by fura-2 assay. Western blot analyses showed alternations in the levels of Bax (pro-apoptotic and Bcl-2 (anti-apoptotic proteins resulting in increased Bax:Bcl-2 ratio in TMZ treated cells. Western blot analyses also detected overexpression of calpain and caspase-3, which cleaved 270 kD α-spectrin at specific sites for generation of 145 and 120 kD spectrin break down products (SBDPs, respectively. However, 1-h pretreatment of cells with 40 μM DXM dramatically decreased TMZ induced apoptosis, decreasing Bax:Bcl-2 ratio and SBDPs. Conclusion Our results revealed an antagonistic effect of DXM on TMZ induced apoptosis in human glioblastoma U87MG cells, implying that treatment of glioblastoma patients with DXM prior to chemotherapy with TMZ might result in an undesirable clinical outcome.

  3. Suppression of the Arboviruses Dengue and Chikungunya Using a Dual-Acting Group-I Intron Coupled with Conditional Expression of the Bax C-Terminal Domain.

    Directory of Open Access Journals (Sweden)

    James R Carter

    Full Text Available In portions of South Asia, vectors and patients co-infected with dengue (DENV and chikungunya (CHIKV are on the rise, with the potential for this occurrence in other regions of the world, for example the United States. Therefore, we engineered an antiviral approach that suppresses the replication of both arboviruses in mosquito cells using a single antiviral group I intron. We devised unique configurations of internal, external, and guide sequences that permit homologous recognition and splicing with conserved target sequences in the genomes of both viruses using a single trans-splicing Group I intron, and examined their effectiveness to suppress infections of DENV and CHIKV in mosquito cells when coupled with a proapoptotic 3' exon, ΔN Bax. RT-PCR demonstrated the utility of these introns in trans-splicing the ΔN Bax sequence downstream of either the DENV or CHIKV target site in transformed Aedes albopictus C6/36 cells, independent of the order in which the virus specific targeting sequences were inserted into the construct. This trans-splicing reaction forms DENV or CHIKV ΔN Bax RNA fusions that led to apoptotic cell death as evidenced by annexin V staining, caspase, and DNA fragmentation assays. TCID50-IFA analyses demonstrate effective suppression of DENV and CHIKV infections by our anti-arbovirus group I intron approach. This represents the first report of a dual-acting Group I intron, and demonstrates that we can target DENV and CHIKV RNAs in a sequence specific manner with a single, uniquely configured CHIKV/DENV dual targeting group I intron, leading to replication suppression of both arboviruses, and thus providing a promising single antiviral for the transgenic suppression of multiple arboviruses.

  4. Differential regulation of spontaneous and immune complex-induced neutrophil apoptosis by proinflammatory cytokines. Role of oxidants, Bax and caspase-3.

    Science.gov (United States)

    Ottonello, Luciano; Frumento, Guido; Arduino, Nicoletta; Bertolotto, Maria; Dapino, Patrizia; Mancini, Marina; Dallegri, Franco

    2002-07-01

    Neutrophil apoptosis represents a crucial step in the mechanisms governing the resolution of neutrophilic inflammation. Several soluble mediators of inflammation modulate neutrophil survival, retarding their apoptosis, whereas neutrophil activation by immune complexes (IC) results in the acceleration of apoptosis. To investigate neutrophil fate at the site of inflammation, we studied the effects of interleukin (IL)-2, IL-6, IL-8, IL-15, GM-CSF, and fMLP on spontaneous and IC-induced neutrophil apoptosis and the mechanisms regulating the survival of these cells. Spontaneous apoptosis was inhibited by GM-CSF, IL-6, and IL-15, but only GM-CSF overturned IC-induced apoptosis. No role of oxidants on the modulation of IC-dependent apoptosis was found. Indeed, fMLP or GM-CSF augmented the IC-dependent oxidative response, whereas the other compounds were ineffective. CGD neutrophils showed low levels of spontaneous apoptosis, but when exposed to IC, underwent a sharp increment of the apoptotic rate in a GM-CSF-inhibitable manner. Conversely, the expression of the proapoptotic protein Bax in 18-h aged neutrophils was down-regulated by GM-CSF, IL-6, and IL-15. Furthermore, IC induced a nearly threefold Bax up-regulation, which was completely reversed only by GM-CSF. Accordingly, the spontaneous activity of caspase-3 was inhibited by GM-CSF, IL-6, and IL-15. Furthermore, IC induced a sharp increment of enzymatic activity, and only GM-CSF inhibited the IC-dependent acceleration. Our results show that apoptosis of resting and IC-activated neutrophils is regulated differently, GM-CSF being the most potent neutrophil antiapoptotic factor. The results also unveil the existence of an oxidant-independent, Bax- and caspase-3-dependent, intracellular pathway regulating neutrophil apoptosis.

  5. Bone Marrow Stromal Cells Promote Neuronal Restoration in Rats with Traumatic Brain Injury: Involvement of GDNF Regulating BAD and BAX Signaling

    Directory of Open Access Journals (Sweden)

    Qin Shen

    2016-02-01

    Full Text Available Background/Aims: To investigate the effects of bone marrow stromal cells (BMSCs and underlying mechanisms in traumatic brain injury (TBI. Methods: Cultured BMSCs from green fluorescent protein-transgenic mice were isolated and confirmed. Cultured BMSCs were immediately transplanted into the regions surrounding the injured-brain site to test their function in rat models of TBI. Neurological function was evaluated by a modified neurological severity score on the day before, and on days 7 and 14 after transplantation. After 2 weeks of BMSC transplantation, the brain tissue was harvested and analyzed by microarray assay. And the coronal brain sections were determined by immunohistochemistry with mouse anti-growth-associated protein-43 kDa (anti-GAP-43 and anti-synaptophysin to test the effects of transplanted cells on the axonal regeneration in the host brain. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL assay and Western blot were used to detect the apoptosis and expression of BAX and BAD. Results: Microarray analysis showed that BMSCs expressed growth factors such as glial cell-line derived neurotrophic factor (GDNF. The cells migrated around the injury sites in rats with TBI. BMSC grafts resulted in an increased number of GAP-43-immunopositive fibers and synaptophysin-positive varicosity, with suppressed apoptosis. Furthermore, BMSC transplantation significantly downregulated the expression of BAX and BAD signaling. Moreover, cultured BMSC transplantation significantly improved rat neurological function and survival. Conclusion: Transplanted BMSCs could survive and improve neuronal behavior in rats with TBI. Mechanisms of neuroprotection and regeneration were involved, which could be associated with the GDNF regulating the apoptosis signals through BAX and BAD.

  6. Bax, Bcl2, and p53 differentially regulate neomycin- and gentamicin-induced hair cell death in the zebrafish lateral line.

    Science.gov (United States)

    Coffin, Allison B; Rubel, Edwin W; Raible, David W

    2013-10-01

    Sensorineural hearing loss is a normal consequence of aging and results from a variety of extrinsic challenges such as excessive noise exposure and certain therapeutic drugs, including the aminoglycoside antibiotics. The proximal cause of hearing loss is often death of inner ear hair cells. The signaling pathways necessary for hair cell death are not fully understood and may be specific for each type of insult. In the lateral line, the closely related aminoglycoside antibiotics neomycin and gentamicin appear to kill hair cells by activating a partially overlapping suite of cell death pathways. The lateral line is a system of hair cell-containing sense organs found on the head and body of aquatic vertebrates. In the present study, we use a combination of pharmacologic and genetic manipulations to assess the contributions of p53, Bax, and Bcl2 in the death of zebrafish lateral line hair cells. Bax inhibition significantly protects hair cells from neomycin but not from gentamicin toxicity. Conversely, transgenic overexpression of Bcl2 attenuates hair cell death due to gentamicin but not neomycin, suggesting a complex interplay of pro-death and pro-survival proteins in drug-treated hair cells. p53 inhibition protects hair cells from damage due to either aminoglycoside, with more robust protection seen against gentamicin. Further experiments evaluating p53 suggest that inhibition of mitochondrial-specific p53 activity confers significant hair cell protection from either aminoglycoside. These results suggest a role for mitochondrial p53 activity in promoting hair cell death due to aminoglycosides, likely upstream of Bax and Bcl2.

  7. Punicalagin attenuated cerebral ischemia-reperfusion insult via inhibition of proinflammatory cytokines, up-regulation of Bcl-2, down-regulation of Bax, and caspase-3.

    Science.gov (United States)

    Yaidikar, Lavanya; Thakur, Santhrani

    2015-04-01

    Punicalagin (PG) is a hydrolysable tannin compound found in Punica granatum L. The purpose of the present work is to explore the neuroprotective mechanism of PG against ischemia-reperfusion (I/R) injury in rat model of middle cerebral artery occlusion (MCAO). Rats were randomly divided into sham, MCAO, and PG-treated groups. PG (15 and 30 mg/kg), the vehicle was administered orally for 7 days prior to MCAO. Rats were anesthetised with ketamine (100 mg/kg/im), xylazine (10 mg/kg/im) and subjected to 2 h occlusion and 22 h reperfusion. The effects of PG on behavioral deficit and infarct volume, the levels of glutamate and calcium as well as the levels of inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) were evaluated. Moreover, the expressions of caspase-3, Bcl-2, and Bax were detected by Western blotting. As compared with MCAO group, PG-treated rats showed dose-dependent reduction in infarct volume and substantial improvement in behavioral deficit. The levels of glutamate, calcium, TNF-α, IL-1β, and IL-6 were restored significantly. The Western blotting results revealed that the expression of Bcl-2 was up-regulated and that of caspase-3, Bax were down-regulated when exposed to PG. From our results, it can be concluded that PG showed an ameliorative effect against cerebral I/R injury in rats through its anti-inflammatory, antioxidant actions besides it inhibits excitotoxicity. It also suppresses apoptosis through regulating, Bcl-2, caspase-3, and Bax protein expressions, perhaps another mechanism by which PG employs its neuroprotective action. PMID:25555468

  8. Multiple doses of erythropoietin impair liver regeneration by increasing TNF-alpha, the Bax to Bcl-xL ratio and apoptotic cell death.

    Directory of Open Access Journals (Sweden)

    Katja Klemm

    Full Text Available BACKGROUND: Liver resection and the use of small-for-size grafts are restricted by the necessity to provide a sufficient amount of functional liver mass. Only few promising strategies to maximize liver regeneration are available. Apart from its erythropoiesis-stimulating effect, erythropoietin (EPO has meanwhile been recognized as mitogenic, tissue-protective, and anti-apoptotic pleiotropic cytokine. Thus, EPO may support regeneration of hepatic tissue. METHODOLOGY: Rats undergoing 68% hepatectomy received daily either high dose (5000 IU/kg bw i.v. or low dose (500 IU/kg bw i.v. recombinant human EPO or equal amounts of physiologic saline. Parameters of liver regeneration and hepatocellular apoptosis were assessed at 24 h, 48 h and 5 d after resection. In addition, red blood cell count, hematocrit and serum EPO levels as well as plasma concentrations of TNF-alpha and IL-6 were evaluated. Further, hepatic Bcl-x(L and Bax protein expression were analyzed by Western blot. PRINCIPAL FINDINGS: Administration of EPO significantly reduced the expression of PCNA at 24 h followed by a significant decrease in restitution of liver mass at day 5 after partial hepatectomy. EPO increased TNF-alpha levels and shifted the Bcl-x(L to Bax ratio towards the pro-apoptotic Bax resulting in significantly increased hepatocellular apoptosis. CONCLUSIONS: Multiple doses of EPO after partial hepatectomy increase hepatocellular apoptosis and impair liver regeneration in rats. Thus, careful consideration should be made in pre- and post-operative recombinant human EPO administration in the setting of liver resection and transplantation.

  9. Involvement of NF-κB and Bcl2/Bax signaling pathways in the apoptosis of MCF7 cells induced by a xanthone compound Pyranocycloartobiloxanthone A.

    Science.gov (United States)

    Mohan, Syam; Abdelwahab, Siddig Ibrahim; Kamalidehghan, Behnam; Syam, Suvitha; May, Koh Sue; Harmal, Nabil Saad Mohammed; Shafifiyaz, Noor; Hadi, A Hamid A; Hashim, Najihah Mohd; Rahmani, Mawardi; Taha, Manal Mohamed Elhassan; Cheah, Shiau-Chuen; Zajmi, Asdren

    2012-08-15

    The plant Artocarpus obtusus is a tropical plant that belongs to the family Moraceae. In the present study a xanthone compound Pyranocycloartobiloxanthone A (PA) was isolated from this plant and the apoptosis mechanism was investigated. PA induced cytotoxicity was observed using MTT assay. High content screening (HCS) was used to observe the nuclear condensation, cell permeability, mitochondrial membrane potential (MMP) and cytochrome c release. Reactive oxygen species formation was investigated on treated cells by using fluorescent analysis. Human apoptosis proteome profiler assays were performed to investigate the mechanism of cell death. In addition mRNA levels of Bax and Bcl2 were also checked using RT-PCR. Caspase 3/7, 8 and 9 were measured for their induction while treatment. The involvement of NF-κB was analyzed using HCS assay. The results showed that PA possesses the characteristics of selectively inducing cell death of tumor cells as no inhibition was observed in non-tumorigenic cells even at 30 μg/ml. Treatment of MCF7 cells with PA induced apoptosis with cell death-transducing signals, that regulate the MMP by down-regulation of Bcl2 and up-regulation of Bax, triggering the cytochrome c release from mitochondria to cytosol. The release of cytochrome c triggered the activation of caspases-9, then activates downstream executioner caspase-3/7 and consequently cleaved specific substrates leading to apoptotic changes. This form of apoptosis was found closely associated with the extrinsic pathway caspase (caspase-8) and inhibition of translocation of NF-κB from cytoplasm to nucleus. The results demonstrated that PA induced apoptosis of MCF7 cells through NF-κB and Bcl2/Bax signaling pathways with the involvement of caspases.

  10. MYC through miR-17-92 Suppresses Specific Target Genes to Maintain Survival, Autonomous Proliferation, and a Neoplastic State

    KAUST Repository

    Li, Yulin

    2014-08-01

    The MYC oncogene regulates gene expression through multiple mechanisms, and its overexpression culminates in tumorigenesis. MYC inactivation reverses turmorigenesis through the loss of distinguishing features of cancer, including autonomous proliferation and survival. Here we report that MYC via miR-17-92 maintains a neoplastic state through the suppression of chromatin regulatory genes Sin3b, Hbp1, Suv420h1, and Btg1, as well as the apoptosis regulator Bim. The enforced expression of miR-17-92 prevents MYC suppression from inducing proliferative arrest, senescence, and apoptosis and abrogates sustained tumor regression. Knockdown of the five miR-17-92 target genes blocks senescence and apoptosis while it modestly delays proliferative arrest, thus partially recapitulating miR-17-92 function. We conclude that MYC, via miR-17-92, maintains a neoplastic state by suppressing specific target genes.

  11. Effect of nitric oxide with different doses on Bcl-2/Bax in spinal dorsal horn in rats induced by formalin%不同剂量的一氧化氮对福尔马林炎性痛大鼠脊髓背角Bcl-2/Bax表达的影响

    Institute of Scientific and Technical Information of China (English)

    未小明; 李宽; 祁文秀

    2011-01-01

    Objective: To investigate the effects of multiple application of different doses of nitric oxide (NO) on Bcl-2/ Bax in spinal dorsal horn induced by formalin. Methods: A succession of 4 d intrathecal injection of NO precursor L-arginine (L-Arg)10 μg/d (low L-Arg group) or 250 μg/d (high L-Arg group) or NOS inhibitor Nω-nitro-L-arginine methylester (L-NAME) 2700 μg/d (L-NAME group) in rats, and normal saline (NS group) was applied as a control, and administration once a day. Then rats were subcutaneously injected formalin (2%, 100 μL) into the right hindpaw, four hours later after formalin injection, Bcl-2 or Bax protein expression were detected with immunocytochemistry and Western Blot. Results: The immunocytochemistry showed the distributions of Bcl-2 and Bax were in both sides of the dorsal horn,especially in superficial laminae, and the expressions of bcl-2 and bax in the ipsilateral side of formalin injection were significantly increased than that in contralateral side of formalin injection in all four groups; the ratio of Bcl-2/Bax with Western-Blot was increased in low L-Arg group compared with normal saline group and was all decreased in high L-Arg group or L-NAME group compared with normal saline group. bcl-2 and bax are two major genes in the regulation of apoptosis, bcl-2 inhibits apoptosis and bax promotes apoptosis. Conclusion: Therefore, in inflammatory pain model, low doses of NO can promote the antiapoptotic gene expression, while high doses of NO and insufficient of NO both can promote pro-apoptotic gene expression, which affect the incidence of inflammatory pain.%目的:探讨多次应用不同剂量的一氧化氮(NO)对福尔马林炎性痛中脊髓背角神经元Bcl-2、Bax表达的影响.方法:连续4 d给大鼠各进行鞘内注射不同剂量的一氧化氮前体左旋精氨酸(L-arginine,L-Arg)10μg/d(低L-Arg组)、250 μg/d(高L-Arg组)或一氧化氮合酶(nitric oxide synthase,NOS)抑制剂Nω-硝基-L

  12. Structural and energetical studies of the adsorption of para and meta-isomers of xylene on pre-hydrated zeolite BaX. Characterization by neutron diffraction and temperature programmed desorption; Etude structurale et energetique de l'adsorption des isomeres para- et meta- du xylene dans la zeolithe BaX prehydratee. Caracterisation par diffraction des neutrons et thermodesorption programmee

    Energy Technology Data Exchange (ETDEWEB)

    Pichon, Ch.

    1999-10-19

    The separation of p-xylene from C{sub 8} aromatics is performed industrially by selective adsorption on zeolitic materials. FAU-type zeolites are currently used for this separation and especially the partially hydrated BaX. The aim of this work is to characterize from a structural (by low temperature neutron powder diffraction) and an energetical (by temperature programmed desorption) point of view, the adsorption of para- and meta- isomers of xylene, for different fillings, as pure substances as well as mixtures, on pre-hydrated zeolite BaX. The influence of the water pre-adsorption on xylene adsorption selectivity is carefully discussed. The crystalline structure of the zeolite BaX (framework and compensation of charge cations) and of the adsorbed phase (water, p- and m-xylene molecules) are completely characterized by neutron diffraction. The location and the distribution of water and xylene molecules on their adsorption sites is especially followed as a function of the filling of the zeolite and of the composition of the adsorbed phase. Microscopic measurements were correlated to the energetical analysis (at a macroscopic level) in order to obtain a consistent description of adsorption phenomenon and to propose a possible origin for adsorption selectivity.

  13. Matrine inhibits diethylnitrosamine-induced HCC proliferation in rats through inducing apoptosis via p53, Bax-dependent caspase-3 activation pathway and down-regulating MLCK overexpression.

    Science.gov (United States)

    Zhang, Xiaolin; Yu, Hao

    2016-01-01

    The proliferation of hepatocellular carcinoma (HCC) cells is one of the leading causes of liver cancer mortality in humans. The inhibiting effects of matrine on HCC cell proliferation have been studied, but the mechanism of that inhibition has not been fully elucidated. Since, apoptosis plays an important role in HCC cell proliferation. We examined the apoptosis-inducing effect of matrine on tumor cells. Western blot analysis of p53, Bax, cleaved caspase-3 and myosin light chain kinase (MLCK) revealed that matrine induced tumor cell apoptosis by controlling anoikis. It activated p53, Bax-dependent caspase-3 and blocked the ECM-integrin mediated cell survival pathway through down-regulating MLCK over-expression in the liver of rats with diethyl nitrosamine (DENA)-induced HCC. Our results suggest that matrine can inhibit the proliferation of HCC cells through inducing tumor cell apoptosis via activation of the p53 pathway and inhibition of MLCK overexpression. Matrine may thus be used as a potentially promising reagent to inhibit HCC cell proliferation and MLCK may be a novel target for the treatment of HCC. PMID:27642320

  14. Levofloxacin increases the effect of serum deprivation on anoikis of rat nucleus pulposus cells via Bax/Bcl-2/caspase-3 pathway.

    Science.gov (United States)

    Yang, Si-Dong; Bai, Zhi-Long; Zhang, Feng; Ma, Lei; Yang, Da-Long; Ding, Wen-Yuan

    2014-12-01

    Levofloxacin, a fluoroquinolone, is a widely-used and effective antibiotic. However, various adverse side effects are associated with levofloxacin. The purpose of this study was to further explore the effects of levofloxacin on rat nucleus pulposus cells (NPCs). Inverted phase-contrast microscopy, flow cytometry and caspase-3 activity assays were used and revealed that serum deprivation induced apoptosis, which was markedly increased by levofloxacin in a dose-dependent manner. Simultaneously, levofloxacin decreased cell binding to type II collagen (COL2). Thus, levofloxacin-induced apoptosis exhibits characteristics of anoikis, the process by which cell death is triggered by separation from the extracellular matrix, which contains COL2. Furthermore, real-time quantitative RT-PCR was used to further confirm that levofloxacin downregulates COL2 expression in a dose-dependent manner. At last, western blot was used to find that levofloxacin increased the ratio of Bax/Bcl-2 and active caspase-3 in a dose-dependent manner. Levofloxacin therefore increases the effects of serum deprivation on anoikis by downregulating COL2 in rat NPCs in vitro via Bax/Bcl-2/caspase-3 pathway. This research provides a novel insight into the mechanisms of levofloxacin-induced toxicity and may potentially lead to a better understanding of the clinical effects of levofloxacin, especially in terms of intervertebral disc degeneration. PMID:25224805

  15. Inotodiol inhabits proliferation and induces apoptosis through modulating expression of cyclinE, p27, bcl-2, and bax in human cervical cancer HeLa cells.

    Science.gov (United States)

    Zhao, Li-Wei; Zhong, Xiu-Hong; Yang, Shu-Yan; Zhang, Yi-Zhong; Yang, Ning-Jiang

    2014-01-01

    Inonotus obliquus is a medicinal mushroom that has been used as an effective agent to treat various diseases such as diabetes, tuberculosis and cancer. Inotodiol, an included triterpenoid shows significant anti-tumor effect. However, the mechanisms have not been well documented. In this study, we aimed to explore the effect of inotodiol on proliferation and apoptosis in human cervical cancer HeLa cells and investigated the underlying molecular mechanisms. HeLa cells were treated with different concentrations of inotodiol. The MTT assay was used to evaluate cell proliferating ability, flow cytometry (FCM) was employed for cell cycle analysis and cell apoptosis, while expression of cyclinE, p27, bcl-2 and bax was detected by immunocytochemistry. Proliferation of HeLa cells was inhibited by inotodiolin a dose-dependent manner at 24h (r=0.9999, pInonotus obliquus inhibited the proliferation of HeLa cells and induced apoptosis in vitro. The mechanisms may be related to promoting apoptosis through increasing the expression of bax and cutting bcl-2 and affecting the cell cycle by down-regulation the expression of cyclin E and up-regulation of p27. The results further indicate the potential value of inotodiol for treatment of human cervical cancer. PMID:24815470

  16. Matrine inhibits diethylnitrosamine-induced HCC proliferation in rats through inducing apoptosis via p53, Bax-dependent caspase-3 activation pathway and down-regulating MLCK overexpression.

    Science.gov (United States)

    Zhang, Xiaolin; Yu, Hao

    2016-01-01

    The proliferation of hepatocellular carcinoma (HCC) cells is one of the leading causes of liver cancer mortality in humans. The inhibiting effects of matrine on HCC cell proliferation have been studied, but the mechanism of that inhibition has not been fully elucidated. Since, apoptosis plays an important role in HCC cell proliferation. We examined the apoptosis-inducing effect of matrine on tumor cells. Western blot analysis of p53, Bax, cleaved caspase-3 and myosin light chain kinase (MLCK) revealed that matrine induced tumor cell apoptosis by controlling anoikis. It activated p53, Bax-dependent caspase-3 and blocked the ECM-integrin mediated cell survival pathway through down-regulating MLCK over-expression in the liver of rats with diethyl nitrosamine (DENA)-induced HCC. Our results suggest that matrine can inhibit the proliferation of HCC cells through inducing tumor cell apoptosis via activation of the p53 pathway and inhibition of MLCK overexpression. Matrine may thus be used as a potentially promising reagent to inhibit HCC cell proliferation and MLCK may be a novel target for the treatment of HCC.

  17. Hsp105 family proteins suppress staurosporine-induced apoptosis by inhibiting the translocation of Bax to mitochondria in HeLa cells

    International Nuclear Information System (INIS)

    Hsp105 (Hsp105α and Hsp105β), major heat shock proteins in mammalian cells, belong to a subgroup of the HSP70 family, HSP105/110. Previously, we have shown that Hsp105α has completely different effects on stress-induced apoptosis depending on cell type. However, the molecular mechanisms by which Hsp105α regulates stress-induced apoptosis are not fully understood. Here, we established HeLa cells that overexpress either Hsp105α or Hsp105β by removing doxycycline and examined how Hsp105 modifies staurosporine (STS)-induced apoptosis in HeLa cells. Apoptotic features such as the externalization of phosphatidylserine on the plasma membrane and nuclear morphological changes were induced by the treatment with STS, and the STS-induced apoptosis was suppressed by overexpression of Hsp105α or Hsp105β. In addition, we found that overexpression of Hsp105α or Hsp105β suppressed the activation of caspase-3 and caspase-9 by preventing the release of cytochrome c from mitochondria. Furthermore, the translocation of Bax to mitochondria, which results in the release of cytochrome c from the mitochondria, was also suppressed by the overexpression of Hsp105α or Hsp105β. Thus, it is suggested that Hsp105 suppresses the stress-induced apoptosis at its initial step, the translocation of Bax to mitochondria in HeLa cells

  18. Matrine inhibits diethylnitrosamine-induced HCC proliferation in rats through inducing apoptosis via p53, Bax-dependent caspase-3 activation pathway and down-regulating MLCK overexpression

    Science.gov (United States)

    Zhang, Xiaolin; Yu, Hao

    2016-01-01

    The proliferation of hepatocellular carcinoma (HCC) cells is one of the leading causes of liver cancer mortality in humans. The inhibiting effects of matrine on HCC cell proliferation have been studied, but the mechanism of that inhibition has not been fully elucidated. Since, apoptosis plays an important role in HCC cell proliferation. We examined the apoptosis-inducing effect of matrine on tumor cells. Western blot analysis of p53, Bax, cleaved caspase-3 and myosin light chain kinase (MLCK) revealed that matrine induced tumor cell apoptosis by controlling anoikis. It activated p53, Bax-dependent caspase-3 and blocked the ECM-integrin mediated cell survival pathway through down-regulating MLCK over-expression in the liver of rats with diethyl nitrosamine (DENA)-induced HCC. Our results suggest that matrine can inhibit the proliferation of HCC cells through inducing tumor cell apoptosis via activation of the p53 pathway and inhibition of MLCK overexpression. Matrine may thus be used as a potentially promising reagent to inhibit HCC cell proliferation and MLCK may be a novel target for the treatment of HCC. PMID:27642320

  19. Insulin-like growth factor-1 protects against prion peptide-induced cell death in neuronal cells via inhibition of Bax translocation.

    Science.gov (United States)

    Park, Yang-Gyu; Jeong, Jae-Kyo; Moon, Myung-Hee; Lee, Ju-Hee; Lee, You-Jin; Seol, Jae-Won; Kim, Shang-Jin; Kang, Seog-Jin; Park, Sang-Youel

    2012-11-01

    Insulin-like growth factor-1 (IGF-1) is one of the most important components of bovine colostrum. It exhibits antiapoptotic and antioxidative activities. Prion diseases are neurodegenerative disorders caused by cell death through mitochondrial dysfunction and increasing generation of reactive oxygen species (ROS). This study examined the protective effect of IGF-1 on residues 106-126 of the cellular prion protein [PrP (106-126)]-mediated mitochondrial neurotoxicity and oxidative stress. In SH-SY5Y human neuronal cells, treatment with PrP (106-126) decreased the cell viability and IGF-1 pretreatment markedly blocked the PrP (106-126)-induced neuronal cell death. IGF-1 inhibited PrP (106-126)-induced intracellular ROS generation and mitochondrial oxidative stress. In addition, IGF-1 blocked the translocation of the Bax protein to the mitochondria induced by PrP (106-126). These results demonstrate that IGF-1 protects neuronal cells against PrP (106-126)-mediated neurotoxicity through an antioxidative effect and blockage of mitochondrial Bax translocation. The results also suggest that regulation of IGF-1 secretion may have a therapeutic potential in the management of mitochondrial dysfunction and oxidative stress-induced neurodegeneration. PMID:22895829

  20. 苦参碱对 HaCaT 细胞 Bcl -2/Bax 和Fas/FasL 的调控%Regulation of Bcl-2/Bax and Fas/FasL by matrine in HaCaT cells

    Institute of Scientific and Technical Information of China (English)

    牟宽厚; 周艳; 韩丹; 穆欣

    2014-01-01

    目的:明确苦参碱对 HaCaT 细胞 Bcl-2/ Bax 和 Fas/ FasL 表达的影响。方法:体外培养HaCaT 细胞,选择第二代细胞对数生长期 HaCaT 细胞作为研究对象,将细胞随机分为4组:苦参碱2 mg/ mL、10 mg/ mL 和50 mg/ mL 3组及对照组(加入相同体积的0.9%盐水),孵育48 h 后,MTT 法测定各浓度下细胞增殖,RT-PCR 检测 Bcl-2/ Bax 和 Fas/ FasL 的表达。结果:与对照组相比,当苦参碱浓度为2 mg/ mL 时,HaCaT 细胞增殖活性无明显变化;Bcl -2、Bax、Fas、FasL 表达也无明显变化( P>0.05)。当苦参碱浓度为10 mg/ mL 时 HaCaT 细胞增殖活性较对照组明显下降(P0.05)。结论:苦参碱能够调控上皮细胞致炎因子的表达,抑制细胞的增殖。%To determine the effect of matrine on Bcl-2/ Bax and Fas/ FasL in keratinocytes in vitro. Methods: Second generation cultured HaCaT cells (logarithmic phase cells) were selected and divided into 4 groups:3 matrine groups (2 mg/ mL, 10 mg/ mL and 50 mg/ mL were used in each group) and the control group (0.9% Natrii Chloride). After 48-hour culture, the proliferation of HaCaT were detected by MTT and the levels of Bcl-2/ Bax and Fas/ FasL were measured by RT-PCR. Results: The viability of HaCaT cells was similar in 2 mg/ mL matrine group and control group (P>0.05). In 10 mg/ mL matrine group the proliferation of the cells was significantly decreased (P<0.001) and the Bcl-2 expression was remarkably reduced (P<0.001), while the expression of Bax, Fas and FasL was significantly increased (P<0.01 and P<0.05, respectively). When the concentration of matrine was increased to 50 mL, the viability and the expression of Bcl-2, Bax, Fas and FasL was similar to the results when 10 mL matrine was used. Conclusion: Matrine can inhibit HaCaT cells proliferation (at 10 mg/ mL or more) and may adjust expression of Bcl-2/ Bax and Fas/FasL in HaCaT cells.

  1. A- and B-site doping effect on physicochemical properties of Sr2‑xBaxMMoO6 (M = Mg, Mn, Fe) double perovskites — candidate anode materials for SOFCs

    Science.gov (United States)

    Zheng, Kun; Świerczek, Konrad

    2016-06-01

    In this work, we evaluate the physicochemical properties of Sr2‑xBaxMMoO6 (M = Mg, Mn, Fe) double perovskites as alternative anode materials for solid oxide fuel cells, for which the effect of substitution of strontium by barium in a full range of compositions is studied. The crystal structure, microstructure, characterization of transport properties (electrical conductivity, Seebeck coefficient) and oxygen content as a function of temperature, as well as chemical stability in oxidizing and reducing conditions are discussed. Fe- and Mo-containing Sr2‑xBaxFeMoO6 oxides show very high total conductivities with values of 100-1000 Sṡcm‑1, while Sr2‑xBaxMgMoO6 present good redox stability.

  2. Data on four apoptosis-related genes in the colonial tunicate Botryllus schlosseri.

    Science.gov (United States)

    Franchi, Nicola; Ballin, Francesca; Manni, Lucia; Schiavon, Filippo; Ballarin, Loriano

    2016-09-01

    The data described are related to the article entitled "Recurrent phagocytosis-induced apoptosis in the cyclical generation change of the compound ascidian Botryllus schlosseri" (Franchi et al., 2016) [1]. Four apoptosis-related genes, showing high similarity with mammalian Bax (a member of the Bcl-2 protein family), AIF1 (apoptosis-inducing factor-1), PARP1 (poly ADP ribose polymerase-1) and IAP7 (inhibitor of apoptosis-7) were identified from the analysis of the trascriptome of B. schlosseri. They were named BsBax, BsAIF1, BsPARP1 and BsIAP7. Here, their deduced amino acid sequence were compared with known sequences of orthologous genes from other deuterostome species together with a study of their identity/similarity. PMID:27294183

  3. Genes and Gene Therapy

    Science.gov (United States)

    ... correctly, a child can have a genetic disorder. Gene therapy is an experimental technique that uses genes to ... or prevent disease. The most common form of gene therapy involves inserting a normal gene to replace an ...

  4. Effects of acupoint versus non-acupoint electroacupuncture on cerebral cortical neuronal Bcl-2,Bax and caspase-3 expression in a rat model of focal cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    Jun Wang; Junming Fan; Yongshu Dong; Xia Huang; Hongxia Zhang

    2008-01-01

    BACKGROUND: Several studies have demonstrated that electroacupuncture by acupoint selection can inhibit cerebral cortical neuronal apoptosis following cerebral ischemia/reperfusion.OBJECTIVE: To validate the effects of electroacupuncture by acupoint selection on the expression level of cortical neuronal anti-apoptotic Bcl-2 protein and the apoptotic executive protein, caspase-3, in rat models of focal cerebral ischemia/reperfusion.DESIGN, TIME AND SETTING: This randomized grouping, neural cell and molecular biology animal experiment was performed at the Laboratory of Pharmacology of Traditional Chinese Medicine and the Laboratory Animal Center of Henan Institute of Traditional Chinese Medicine between November 2006 and May 2007.MATERIALS: Atotal of 40 healthy male adult Sprague-Dawley rats were randomly and evenly divided into four groups: sham-operated, model, electroacupuncture and non-acupoint control. G6895 electro-acupuncture instruments were purchased from Shanghai Huayi Instrument Factory, China. Caspase-3, Bcl-2 and Bax kits were provided by Wuhan Boster Bioengineering Co., Ltd., China.METHODS: Middle cerebral artery occlusion was induced in the model, electroacupuncture and non-acupoint groups. In the electroacupuncture group, the acupoints Jianyu (LI15), Waiguan (SJ5), Biguan (ST31), and Zusanli (ST36) were given electroacupuncture. In the non-acupoint control group, at each time point (immediately after ischemia and after reperfusion, or 2 hours after reperfusion), electroacupuncture was performed at the midpoints of Tianquan (PC2)-Quze (PC 3) line, Quze (PC 3)-Ximen (PC4) line, Zuwuli (LRlO)-Yinbao (LRg) line, and Xiguan (LR7)-Zhongdu (LR6) line. Electroacupuncture parameters were set with a continuous wave with a frequency of 10 Hz, wave width 0.6 ms, voltage 1.5-3.0 V, and a duration of 10 minutes. The sham-operated and model groups received only animal fixation without electroacupuncture procedure.MAIN OUTCOME MEASURES: Five rats were selected from

  5. Effects of human interleukin 10 gene transfer on the expression of Bcl-2 Bax and apoptosis of hepatocyte in rats with acute hemorrhagic necrotizing pancreatitis

    Institute of Scientific and Technical Information of China (English)

    GU Jun-chao; WANG Yu; ZHANG Zhong-tao; XUE Jian-guo; LI Jian-she; ZHOU Yan-zhong

    2005-01-01

    @@ Acute necrotising pancreatitis is characterized by inflammatory and necrotic events, which follow the initial intra-acinar injury involving enzyme activation, and disruption of the acinar cytoskeleton.1 At present, apoptosis has become a hot topic in many kinds of disease.

  6. Caspase-3和bax在视网膜母细胞瘤中的表达%Expression of caspase-3 and bax gene protein in retinoblastoma

    Institute of Scientific and Technical Information of China (English)

    孙红; 惠延年; 王立勤; 马吉献

    2003-01-01

    目的: 观察凋亡及凋亡调控基因caspase-3/bax在视网膜母细胞瘤(retinoblastoma, RB)中的表达及与凋亡的相关性. 方法: 收集35例RB标本,对其分别进行caspase-3和bax免疫组织化学染色,观察表达情况及染色强度. 结果: Caspase-3及bax在未分化型(n=15)分别有较好的表达(11/12例),caspase-3及bax在分化型(n=20)中也有较好的表达(17/18例). 正常视网膜组织中无caspase-3及bax的表达. 结论: 凋亡在RB中是存在的,caspase-3及bax在RB的发生发展中起重要作用.

  7. Resveratrol Reverses Cadmium Chloride-induced Testicular Damage and Subfertility by Downregulating p53 and Bax and Upregulating Gonadotropins and Bcl-2 gene Expression

    OpenAIRE

    Eleawa, Samy M.; ALKHATEEB, Mahmoud A; ALHASHEM, Fahaid H; BIN-JALIAH, Ismaeel; SAKR, Hussein F; ELREFAEY, Hesham M; ELKARIB, Abbas O; ALESSA, Riyad M; HAIDARA, Mohammad A; Shatoor, Abdullah S.; KHALIL, Mohammad A

    2014-01-01

    This study was performed to investigate the protective and therapeutic effects of resveratrol (RES) against CdCl2-induced toxicity in rat testes. Seven experimental groups of adult male rats were formulated as follows: A) controls+NS, B) control+vehicle (saline solution of hydroxypropyl cyclodextrin), C) RES treated, D) CdCl2+NS, E) CdCl2+vehicle, F) RES followed by CdCl2 and M) CdCl2 followed by RES. At the end of the protocol, serum levels of FSH, LH and testosterone were measured in all gr...

  8. Change in expression of apoptosis genes after hyperthermia, chemotherapy and radiotherapy in human colon cancer transplanted into nude mice

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To investigate the change in expression of p53, Bcl-2, and Bax genes in human colon cancer cells transplanted into nude mice after hyperthermia,chemotherapy, radiotherapy, thermochemotherapy,thermoradiotherapy and thermochemoradiotherapy.METHODS: Human colon cancer cell line (HT29)was transplanted into the hind limbs of nude mice.Under laboratory simulated conditions of hyperthermia (43℃, 60 min), the actual radiation doses and doses of mitomycin C (MMC) were calculated in reference to the clinical radiotherapy for human rectal cancer and chemotherapy prescription for colon cancer. The mice were divided into 6 groups according to the treatment approaches: hyperthermia, chemotherapy,radiotherapy, thermochemotherapy, thermoradiotherapy,and thermochemoradiotherapy. The mice were sacrificed at different time points and the tumor tissue was taken for further procedures. The morphologic changes in membrane, cytoplasm and nuclei of tumor cells of p53, Bcl-2, and Bax after treatment, were observed by immunohistochemistry staining.RESULTS: All of the six treatment modalities downregulated the expression of p53, Bcl-2 and up-regulated the expression of Bax at different levels. The combined therapy of hyperthermia, with chemotherapy, and/or irradiation showed a greater effect on down-regulating the expression of p53 (0.208 ± 0.009 vs 0.155 ± 0.0115,P < 0.01) and Bcl-2 (0.086 ± 0.010 vs 0.026 ± 0.0170,P < 0.01) and up-regulating Bax expression (0.091 ±0.0013 vs 0.207 ±0.027, P < 0.01) compared with any single therapy.CONCLUSION: Hyperthermia enhances the effect of radio- and chemotherapy on tumors by changing the expression of apoptosis genes, such as p53, Bcl-2 and Bax.

  9. Preparation of BaxSr1-xTiO3 Functional Ceramic Film by Liquid Self-Assembly Technology%液相自组装制备BaxSr1-xTiO3功能陶瓷薄膜

    Institute of Scientific and Technical Information of China (English)

    谈国强; 程蕾; 苗鸿雁; 王艳

    2011-01-01

    以Sr(NO3)2,Ba(NO3)2,(NH4)2TiF6和H3BO3作为原料,采用自组装单分子膜(self-assembled monolayers,SAMs)技术以及利用紫外光修饰技术对十八烷基三氯硅烷(C18H37SiCl3,OTS)单分子膜进行官能团改性,在氧化铟锡(indium tin oxide,ITO)玻璃基板上成功制备了BaxSr1-xTiO3功能陶瓷薄膜.通过动态/静态接触角仪测量了SAMs功能化的ITO玻璃基板表面与水的接触角,探讨功能化ITO玻璃基板在紫外光照射前后的润湿情况.通过X射线衍射、能量色散光谱和扫描电子显微镜等测试方法分析了制备的BaxSr1-xTiO3功能陶瓷薄膜的物相组成、微区结构和形貌.结果表明:改性的SAMs功能化ITO玻璃基板在50℃的前驱溶液中沉积18h后,在600℃煅烧晶化2h,可以成功制备出纯相BaxSr1-xTiO3功能陶瓷薄膜,薄膜的颗粒均一,形貌均匀.%Using strontium nitrate, barium nitrate, ammonium hexafluorotitanate and boric acid as raw materials, the functional groups of octadecyltrichlorosilane (C18H37SiCl3, OTS) monolayer were modified via self-assembled monolayers (SAMs) technology and ultraviolet modification technique, and the BaxSr1-xTiO3 functional ceramic film on the indium tin oxide (ITO) glass substrate was prepared successfully. The contact angle between the surface of the SAM functionalized ITO glass substrate and water was measured by dynamic/static contact angle instrument in order to study the wetting conditions of the substrates before and after ultraviolet light irradiation. The phase composition, microstructure and morphology of the prepared BaxSr1-xTiO3 functional ceramic thin films were analyzed by X-ray diffraction, energy dispersive spectroscopy and scanning electron microscopy. The results indicate that pure phase BaxSr1-xTiO3 functional ceramic film can be prepared on the SAMs functionalized ITO glass substrate successfully after depositing on the surface of ITO glass substrate in the precursor at 50 ℃ for 18h, and then crystallized

  10. Dielectric dispersion of BaxSr1-xTiO3 thin film with parallel-plate and coplanar interdigital electrodes

    Science.gov (United States)

    Zhang, Xiao-Yu; Song, Qing; Xu, Feng; Sheng, Su; Wang, Peng; Ong, C. K.

    2009-03-01

    Ferroelectric BaxSr1-xTiO3 (BST) thin films with x = 0.25 and 0.5 were grown by pulsed laser deposition on single crystal LaAlO3 and Pt/Ti/SiO2/Si substrates, respectively. Capacitors were then fabricated from the BST thin films based on coplanar interdigital electrodes (CIEs) and parallel-plate electrodes (PPEs). The dielectric properties of the BST film with CIE and PPE were investigated and compared over a wide frequency range from 100 Hz to 10 GHz. The dielectric dispersion in PPE configuration, caused by the interfacial polarization in film/electrode interfaces, exhibited a strong dependence on frequency. However, the permittivity ɛCIE in CIE configuration shows a gentle variation with the frequency indicating interfacial polarization substantially suppressed. The influence upon the dielectric properties of the columnar BST grains due to the use of different forms of electrodes was discussed.

  11. Influence of Tanshinone IIa on heat shock protein 70, Bcl-2 and Bax expression in rats with spinal ischemia/reperfusion injury

    Institute of Scientific and Technical Information of China (English)

    Li Zhang; Weidong Gan; Guoyao An

    2012-01-01

    Tanshinone IIa is an effective monomer component of Danshen, which is a traditional Chinese medicine for activating blood circulation to dissipate blood stasis. Tanshinone IIa can effectively improve brain tissue ischemia/hypoxia injury. The present study established a rat model of spinal cord ischemia/reperfusion injury and intraperitoneally injected Tanshinone IIa, 0.5 hour prior to model establishment. Results showed that Tanshinone IIa promoted heat shock protein 70 and Bcl-2 protein expression, but inhibited Bax protein expression in the injured spinal cord after ischemia/reperfusion injury. Furthermore, Nissl staining indicated a reduction in nerve cell apoptosis and fewer pathological lesions in the presence of Tanshinone IIa, compared with positive control Danshen injection.

  12. Dielectric dispersion of BaxSr1-xTiO3 thin film with parallel-plate and coplanar interdigital electrodes

    International Nuclear Information System (INIS)

    Ferroelectric BaxSr1-xTiO3 (BST) thin films with x = 0.25 and 0.5 were grown by pulsed laser deposition on single crystal LaAlO3 and Pt/Ti/SiO2/Si substrates, respectively. Capacitors were then fabricated from the BST thin films based on coplanar interdigital electrodes (CIEs) and parallel-plate electrodes (PPEs). The dielectric properties of the BST film with CIE and PPE were investigated and compared over a wide frequency range from 100 Hz to 10 GHz. The dielectric dispersion in PPE configuration, caused by the interfacial polarization in film/electrode interfaces, exhibited a strong dependence on frequency. However, the permittivity εCIE in CIE configuration shows a gentle variation with the frequency indicating interfacial polarization substantially suppressed. The influence upon the dielectric properties of the columnar BST grains due to the use of different forms of electrodes was discussed.

  13. Characteristics of silicon-based BaxSr1-xTiO3 thin films prepared by a sol-gel method

    International Nuclear Information System (INIS)

    Silicon-based BaxSr1-xTiO3 (BST) thin films have been prepared by a sol-gel method with rapid thermal annealing (RTA) processes. Phase structure of the films has been investigated by x-ray diffraction. Atomic force microscopy studies reveal a dense and smooth surface of the sol-gel prepared films. Microstructure and electrical properties of the BST films can be affected by the substrate and the annealing process. RTA method is found to be very efficient to improve the electrical properties of the films. Dielectric constant and dielectric loss of the BST films at 100 kHz are 230 and 0.02, respectively. Leakage current density of the BST capacitors is 1.6x10-7A cm-2 at 3 V. (author)

  14. Diffuse phase transition and high-temperature dielectric relaxation study on (Bi0.5Na0.5)1-xBaxTiO3 ceramics

    Science.gov (United States)

    Chen, Feng; Liu, Qiu-Xiang; Tang, Xin-Gui; Jiang, Yan-Ping; Yue, Jing-Long; Li, Jin-Kai

    2016-09-01

    Lead free (Bi0.5Na0.5)1-xBaxTiO3 (x=0.6, 0.7, 0.8, 0.9) ferroelectric ceramics were synthesized by the traditional solid state reaction method. Sintering was carried out at 1200 °C for 2 h in air atmosphere. The structural, microstructure and Ferroelectric of ceramics were investigated. In dielectric studies, a diffuse phase transition was exhibited and a dielectric relaxation behavious was observed at high temperature range. Impedance analysis characterized grain and grain boundaries resistivities of the ceramics and calculated activation energy and the activation energy for conduction. Polaron theory indicates that the relaxation of the samples at high temperatures was associated with the hopping ions caused by oxygen vacancies.

  15. AC Conductivity and Dielectric Relaxation Behavior of Sol-gel BaxSr1-xTiO3 Thin Films

    Institute of Scientific and Technical Information of China (English)

    Ala'eddin A. Saif; P. Poopalan

    2011-01-01

    BaxSr1-xTiO3 sol-gel thin films with x--0.5, 0.7 and 0.8 have been fabricated as AI/BST/Pt capacitor. The AC conductivity and dielectric properties over a frequency rang of 10 Hz and I MHz have been studied in order to explore the ion dynamics and relaxation mechanisms in the films. The frequency dependent conductivity plots show three regions of conduction processes. Dielectric results show that ε' at low frequencies increases as Sr content decreases, whereas at high frequencies, it shows opposite variation, which is attributed to the dipole dynamics. The electric modulus plots reveal the relaxation peaks which are not observed in the ε" plots and the contribution of the grains, grain boundaries and electrode to the relaxation mechanisms.

  16. Study on the Effects of Losartan on Cardiomyocyte Apoptosis and Gene Expression After Ischemia and Reperfusion in vivo in Rats

    Institute of Scientific and Technical Information of China (English)

    ZHANG Dongqing; YANG Liming; LIU Zhengxiang; MI Shizan

    2000-01-01

    In order to study the effects of losartan on cardiomyocyte apoptosis following ischemia (0.5 h) and reperfusion (48 h) in vivo and bcl-2 and bax gene expression, TUNEL staining method, immunohistochemistry and in situ hybridization histochemistry (ISHH) were used to monitor the apoptotic cells, mRNA and protein of gene expression, respectively. Image processing system was used to quantitively dispose the positive metric substance of both immunohistochemistry and ISHH through the average optical density (OD) value. The number of the apoptotic cells were 38±9 (control group), 0-1 (sham operation group) and 9±4 (losartan-treated group) in each visual field respectively with the difference among the groups being significant (P<0.001). OD values of bcl-2 (ISHH) were 0.07425±0.02029 (control group), 0.05961±0.009932 (sham operation group) and 0. 07619±0.01445 (losartan-treated group) respectively,while OD values of bcl-2 (immunohistochemistry) were 0.1374 ±0.01367 (control group),0.08510±0.01862 (sham operation group) and 0.1252±0.02064 (losartan-treated group), bcl-2gene expression was increased significantly in the control group and losartan-treated group as compared with sham operation group (P<0.05). OD value of bax (immunohistochemistry) was09727± 0.02230 (control group), 0.06182±0.01430 (sham operation group) and 0.06213 ±0.01420 (losartan-treated group), bax gene expression was decreased very significantly in losartan-treated group and sham operation group as compared with control group (P<0. 001). Bcl-2/bax ratio was 1.413 (control group), 1.376 (sham operation group) and 2.016 (losartan-treated group) respectively. The results indicated that losartan might inhibit cardiomyocyte apoptosis following ischemia and reperfusion. The mechanism might be that bax gene expression was inhibited to increase bcl-2/bax ratio.

  17. Common Patterns of Bcl-2 Family Gene Expression in Two Traumatic Brain Injury Models

    OpenAIRE

    Strauss, Kenneth I.; NARAYAN, RAJ K.; Raghupathi, Ramesh

    2004-01-01

    Cell death/survival following traumatic brain injury (TBI) may be a result of alterations in the intracellular ratio of death and survival factors. Bcl-2 family genes mediate both cell survival and the initiation of cell death. Using lysate RNase protections assays, mRNA expression of the anti-cell death genes Bcl-2 and Bcl-xL, and the pro-cell death gene Bax, was evaluated following experimental brain injuries in adult male Sprague-Dawley rats. Both the lateral fluid-percussion (LFP) and the...

  18. Magnetic and thermoelectric properties of the ternary pseudo-hollandite BaxCr5Se8 (0.5 < x < 0.55) solid solution.

    Science.gov (United States)

    Lefèvre, Robin; Berthebaud, David; Bux, Sabah; Hébert, Sylvie; Gascoin, Franck

    2016-07-26

    The structure of Ba0.5Cr5Se8 has been recently resolved, and its thermoelectric and magnetic properties have been studied. A ZT of 0.12 was found at around 800 K. Here, we report a study on the pseudo-hollandite BaxCr5Se8 solid-solution with 0.5 ≤ x ≤ 0.55 and its thermoelectric and magnetic properties. There is no significant impact either on the cell parameters depending on the cation content or on the magnetic properties. However, thermoelectric properties are radically changed depending on x content. While the low thermal conductivity, around 0.8 W m(-1) K(-1), remains similar for all samples, a respective increase and decrease of the resistivity and the Seebeck coefficient are observed with increasing Ba content. The maximum Seebeck coefficient is found with Ba0.5Cr5Se8 at around 635 K with 315 μV K(-1), and the Seebeck coefficient then decreases and is correlated with an activation of minority charge carriers confirmed by Hall measurements. A similar but steeper behavior is observed for the Ba0.55Cr5Se8 temperature dependence plot at around 573 K. Finally, the best thermoelectric performances are found using the lowest content of Ba, unlike when x tends to 0.55, ZT approaches a tenth of the initial best value. BaxCr5Se8 compounds are antiferromagnetic with TN = 58 K. A large peak in thermal conductivity is observed around the antiferromagnetic transition for all stoichiometry.

  19. The ethanol extract of Scutellaria baicalensis and the active compounds induce cell cycle arrest and apoptosis including upregulation of p53 and Bax in human lung cancer cells

    International Nuclear Information System (INIS)

    Despite a lack of scientific authentication, Scutellaria baicalensis is clinically used in Chinese medicine as a traditional adjuvant to chemotherapy of lung cancer. In this study, cytotoxicity assays demonstrated that crude ethanolic extracts of S. baicalensis were selectively toxic to human lung cancer cell lines A549, SK-LU-1 and SK-MES-1 compared with normal human lung fibroblasts. The active compounds baicalin, baicalein and wogonin did not exhibit such selectivity. Following exposure to the crude extracts, cellular protein expression in the cancer cell lines was assessed using 2D gel electrophoresis coupled with MALDI-TOF-MS/Protein Fingerprinting. The altered protein expression indicated that cell growth arrest and apoptosis were potential mechanisms of cytotoxicity. These observations were supported by PI staining cell cycle analysis using flow cytometry and Annexin-V apoptotic analysis by fluorescence microscopy of cancer cells treated with the crude extract and pure active compounds. Moreover, specific immunoblotting identification showed the decreased expression of cyclin A results in the S phase arrest of A549 whereas the G0/G1 phase arrest in SK-MES-1 cells results from the decreased expression of cyclin D1. Following treatment, increased expression in the cancer cells of key proteins related to the enhancement of apoptosis was observed for p53 and Bax. These results provide further insight into the molecular mechanisms underlying the clinical use of this herb as an adjuvant to lung cancer therapy. - Research highlights: → Scutellaria baicalensis is a clinical adjuvant to lung cancer chemotherapy in China. → Scutellaria ethanol extracts selectively toxic to A549, SK-LU-1 and SK-MES-1. → Baicalin, baicalein and wogonin were toxic to all lung cancer cell lines. → Proteomics identified increased p53 and BAX in response to Scutellaria extracts.

  20. Differential effects of two pro-apoptotic members of the Bcl-2 gene family on murine bone quality

    Science.gov (United States)

    Wise, Lisa Marie

    Bax and Hrk are pro-apoptotic members of the Bcl-2 gene family. Both Bax and Hrk have been previously implicated in ovarian cell survival. Effects on bone cells have also been studied in several members of the Bcl-2 gene family; thus, the focus of this work was to characterize the bone quality of mice deficient in Bax or Hrk. Bone quality of various age groups (3, 6, 12, 6 and 22 months) of Bax-knockout (KO) and Hrk-KO female mice were compared to age-matched control female mice. Additional groups of 6-month mice were ovariectomized (OVX) to determine whether effects are dependent on ovarian function. Dual energy x-ray absorptiometry was performed on all mice to determine bone mineral density (BMD). To evaluate bone mechanical properties, 3-point bending, torsion testing and femoral neck fracture were performed on femora, while compression was performed on individual vertebrae. Mechanical properties were rationalized through evaluation of structural (strut analysis, micro computed tomography), remodeling (histomorphometry, osteoclast staining) and material (back-scattered electron imaging, x-ray diffraction) properties. Aged Bax-KO mice do not experience the loss in BMD, bone mechanics and trabecular bone structural properties typically observed with age. Enhanced ovarian cell numbers in Bax-KO mice likely indirectly leads to this enhanced bone phenotype. Ovariectomy results in the loss of the enhanced trabecular bone phenotype, but does not affect the cortical bone phenotype. As such, cortical bone may be protected from typical OVX effects due to sustained osteoblast function in Bax-KO mice. By contrast, young Hrk-KO mice exhibit higher BMD and trabecular bone structural properties compared to control mice, coupled with a compromised mechanical integrity. This subtle transient osteopetrotic-like phenotype is likely influenced by a potentially augmented osteoblast survival, albeit with a compromised activity. This osteopetrotic-like phenotype, and the effect of Hrk

  1. Adsorption of xylene para- and meta- isomers in NaX and BaX zeolites. Study of properties-structure relations; Adsorption des isomeres para- et meta- du xylene dans les zeolithes NaX et BaX. Etude des relations proprietes-structure

    Energy Technology Data Exchange (ETDEWEB)

    Descours, A.

    1997-02-14

    The separation of para-xylene from C8 aromatics is performed industrially bu adsorption process on zeolitic molecular sieves. The sorption properties of these zeolites are strongly linked to their structure, and their comprehension require an accurate knowledge of the interactions between sorbate molecules and zeolitic structure. The aim of this work is to characterise from a structural point of view the adsorption of para- and meta-xylenes in BaX and NaX zeolites. The former is selective for para-xylene, and the latter has not selective properties for para- and meta-isomers of xylene. For each zeolite, the adsorption of pure para-xylene and meta-xylene or a mixture of the two isomers, is investigated as a function of coverage. Powder neutron diffraction is used to determine the crystalline structure of these zeolites and the different crystallographic adsorption sites of the molecules. The influence of coverage on sorbate-sorbent and sorbate-sorbate interactions is investigated. Infrared spectroscopy allows to determine the chemical environment of the sorbate molecules at low coverage or when the coverage increases, and is particularly effective for the study of the binary mixture of xylenes. This study is performed by sorbing a mixture of xylene isomers, or by sorbing these isomers successively. Infrared studies and crystallographic analysis are compared in order to get a consistent description of adsorption mechanism of xylene isomers for both zeolites as a function of coverage. The role of coverage, of cation type, an the presence of the two xylene isomers is the super-cages is essential. For both zeolites, the increase of coverage actually leads to steric hindrances between sorbed molecules and molecular rearrangements. These reorganizations are connected to the cationic distribution of NaX and BaX zeolites. The sorbed molecules are connected to the cationic distribution of NaX and BaX zeolites. The sorbed molecules are particularly confined in BaX zeolite

  2. 中度低温体外循环后大鼠海马bcl-2和bax的表达与神经元凋亡%Hippocampal bcl-2 and bax expressions and neuronal apoptosis after moderate hypothermic cardiopulmonary extracorporeal circulation in rats

    Institute of Scientific and Technical Information of China (English)

    张挺杰

    2005-01-01

    BACKGROUND: Hippocampus injury is wildly believed to involve in neurocognitive dysfunction; the establishment of a rat model of cardiopulmonary bypass(CPB) allows us to investigate the mechanism of CPB-related hippocampus injury.OBJECTIVE: To investigate the effects of moderate hypothermic CPB with a hemodilution on hippocampal bcl-2 and bax gene expression and neuronal apoptosis in rats.DESIGN: A randomized group division study based on the experimental animals.SETTING: Department of anesthesiology in a university hospital.MATERIALS: Thirty Sprague-Dawley (SD) rats were randomly divided into two groups, CPB group and sham-CPB group with 15 rats in each group.METHODS: Total 15 rats of CPB group were subjected to 60-minute moderate hypothermic nonpulsatile CPB using a peristaltic pump and a membrane oxygenator. The CPB circuit was primed with approximately 20 mL 1:1crystaloid-colloid liquid, while another 15 rats of sham-CPB group underwent identical anesthetic and surgical procedures(including cannulation) except CPB itself. At 1 hour post-CPB, six rats in each group were decapitated, and hippocampi were removed, homogenized, and processed for apoptotic gene ( bcl-2 and bax) mRNAs detection. Reverse transcriptase polymerase chain reaction(RT-PCR) is used to detect expression of mRNA by comparing the PCR product of bcl-2 or bax to those of β-actin housekeeping gene. Immunohistochemistry is used to detect bcl-2 and bax protein expressions and terminal deoxynucleiotidyl transferase-mediated dUTP-biotion nick end labeling(TUNEL) staining method was used to detect neuronal apoptosis at 6 hours post-CPB ( n = 6 in each group) . The protein expression was quantitated as percentage of the positively stained area in the total stained. In addition, hippocampal neuronal ultrastructures were studied by electron microscopy at 6 hours post-CPB( n = 3 in each group).ronal apoptosis and ultrastructure changes between the two groups.RESULTS: At 1 hour post-CPB, the expressions of

  3. Hesperetin inhibit adipocyte differentiation and enhance Bax- and p21-mediated adipolysis in human mesenchymal stem cell adipogenesis.

    Science.gov (United States)

    Subash-Babu, Pandurangan; Alshatwi, Ali A

    2015-03-01

    We aimed to explore the antiadipogenic and adipolysis effect of hesperetin in human mesenchymal stem cells (hMSCs)-induced adipogenesis. IC50 value of hesperetin was higher for hMSCs such as 149.2 ± 13.2 μmol for 24 h and 89.4 ± 11.4 μmol in 48 h, whereas in preadipocytes was 87.6 ± 9.5 μmol and 72.4 ± 5.6 μmol in 24 h and 48 h, respectively. Hesperetin treatment (5, 10, and 20 μmol) to adipogenesis-induced hMSCs (Group 1) and preadipocytes (Group 2) resulted in a significantly (p p21 expression in Group 2 compared to untreated preadipocytes. hMSCs cultured in adipogenic medium along with hesperetin significantly inhibited adipocyte differentiation and increased the proapoptotic gene expression levels in preadipocyte. Our result indicates the antiadipogenic and adipolysis effects of hesperetin.

  4. 雌激素对大鼠胸腺细胞凋亡及Bcl-2、Bax表达的影响%Effects of estrogen on apoptosis and expression of Bcl-2 and Bax in rat thymus

    Institute of Scientific and Technical Information of China (English)

    李雅娜; 孙研; 崔春红; 殷彦君

    2011-01-01

    目的:探讨苯甲酸雌二醇对大鼠胸腺Bcl-2和Bax表达及细胞凋亡的影响及其机制.方法:雌性大鼠行卵巢切除术,给予苯甲酸雌二醇后,观察胸腺指数的变化,Hochest33342荧光染色及透射电镜标本观察胸腺细胞凋亡情况,免疫组织化学检测胸腺组织中Bcl-2和Bax的表达情况,原位杂交技术检测Bcl-2、Bax m RNA的表达情况.结果:双侧卵巢切除组大鼠胸腺指数较假手术组增加,双侧卵巢切除+雌激素组大鼠胸腺指数较双侧卵巢切除组减小;假手术组和双侧卵巢切除组大鼠胸腺组织中以正常胸腺细胞为主,偶见凋亡细胞或凋亡小体,双侧卵巢切除+雌激素组可见较多凋亡细胞和凋亡小体;双侧卵巢切除+雌激素组大鼠胸腺组织中Bcl-2表达较双侧卵巢切除组和假手术组增高明显降低,而Bax表达呈现相反趋势;Bcl-2 mRNA、Bax mRNA的表达与Bcl-2、Bax的表达呈一致性.结论:雌激素可以降低大鼠胸腺指数,抑制胸腺组织中Bcl-2的表达,促进Bax的表达,从而诱导大鼠胸腺细胞凋亡,促进雌性大鼠胸腺退化.%Objective-. To explore the effects of estrogen on the apoptosis and the expression of Bcl-2 and Bax in rat thymus. Methods-. The rats performed with ovariectomy were injected with estradiol benzoate. Thymus was obtained 7 days after the injection. Thymic indexes were measured. Apoptosis of the thymus was detected after Hochest 33342 staining and examined under an electron microscope. The expression of Bcl-2 and Bax in the thymus was detected with a immunohistochemical method. The expression of Bcl-2 mRNA and Bax mRNA in the thymus was detected by in situ hybridization. The test was taken in statistical treatment. Results: The thymus quality index in ovariectomy group was higher than that in the model control group. The thymus quality index of rats injected with estradiol benzoate was reduced respectively. Apoptotic cells and apoptotic bodies were found in the

  5. Effect of myocardial reperfusion on cardiocyte apoptosis and expression of bcl-2, bax and caspase-3 in rats with depression%心肌再灌注对抑郁大鼠心肌细胞凋亡以及bcl-2、bax和caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    刘淑珍; 尤鑫; 熊小栓; 刘兴德

    2012-01-01

    apoplolic cardiomyocyles were delecled by in siLu TdT - media-led dUTP nick end labeling (TUNEL) melhod, and ihe expression of bcl -2, bax and caspase - 3 was delemined by ihe melhods of immunohislochemislry and reverse Iranscriplion polymerase chain reaction ( RT - PCR) . RESULTS; Compared wilh group A and group B, ihe numbers of apoplolic cardiomyocyles in group C and group D were significantly increased (P < 0. 01) , and ihe expression of bcl - 2, bax and caspase - 3 in group C and group D was also significantly increased ( P < 0. 01). No significant difference between group A and B was observed. Compared wilh group C, the number of apoplolic cardiomyocyles in group D was significantly increased (P < 0. 05). The gene expression of bcl -2 in group D was decreased significantly ( P < 0. 05 ) , while the gene expression of bax and caspase - 3 in group D was significantly increased ( P < 0. 05 ) . CONCLUSION; Myocardial reperfusion increases apoptosis in ischemic cardiomyocyles in the rals with depression. The mechanisms may be associated with up - regulaling the gene expression of bax and caspase - 3 while down - regulaling bcl - 2 expression.

  6. Ginkgo biloba extract mitigates liver fibrosis and apoptosis by regulating p38 MAPK, NF-κB/IκBα, and Bcl-2/Bax signaling

    Directory of Open Access Journals (Sweden)

    Wang YY

    2015-12-01

    Full Text Available Yuanyuan Wang, Rong Wang, Yujie Wang, Ruqin Peng, Yan Wu, Yongfang Yuan Department of Pharmacy, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China Background: Liver fibrosis is the consequence of diverse liver injuries and can eventually develop into liver cirrhosis. Ginkgo biloba extract (GBE is an extract from dried ginkgo leaves that has many pharmacological effects because of its various ingredients and has been shown to be hepatoprotective. Purpose and methods: Aimed to investigate the underlying protective mechanisms of GBE on carbon tetrachloride (CCl4-induced liver fibrosis in rats. Male Sprague Dawley rats were randomly divided into four groups: control group (C, model group (M, low-dose group (L, and high-dose group (H. Liver fibrosis was induced by CCl4 groups M, L, and H: group C was administered saline. In addition, GBE at different doses was used to treat groups L and H. Results: The results of hematoxylin and eosin staining, Masson’s trichrome staining, a liver function index, and a liver fibrosis index showed that GBE application noticeably mitigated fibrosis and improved the function of the liver. The western blotting and immunohistochemistry analyses indicated that GBE reduced liver fibrosis not only by inhibiting p38 MAPK and NF-κBp65 via inhibition of IκBα degradation but also by inhibiting hepatocyte apoptosis via downregulation of Bax, upregulation of Bcl-2, and subsequent inhibition of caspase-3 activation. Inflammation-associated factors and hepatic stellate cell (HSC-activation markers further demonstrated that GBE could effectively inhibit HSC activation and inflammation as a result of its regulation of p38 MAPK and nuclear factor-kappa B/IκBα signaling. Conclusion: Our findings indicated a novel role for GBE in the treatment of liver fibrosis. The potential mechanisms may be associated with the following signaling pathways: 1 the p38 MAPK

  7. 血糖波动对2型糖尿病大鼠认知功能及海马Bax/Bcl-2的影响%The Effect of Glucose Fluctuate on the Cognitive Function and the Bax/Bcl-2 in Hippocampal of Rats with Type 2 Diabetes Mellitus

    Institute of Scientific and Technical Information of China (English)

    徐志伟; 寿旗扬; 李守业; 蔡月琴; 刘琼; 徐文聃; 洪春兰; 王辉

    2013-01-01

    [目的]探讨血糖波动对糖尿病大鼠认知功能和海马Bax/Bcl-2的影响.[方法]取45只雄性大鼠禁食不禁水16h,用链脲佐菌素30 mg·kg-1加高脂饲料诱导2型糖尿病大鼠模型,挑选出糖尿病大鼠30只,分成糖尿病模型组(Diabetes Model,M)、持续高血糖糖尿病组(High Glucose Diabetes Sustain Model,MS)、血糖波动糖尿病组(Glucose Fluctuate Diabetes Model,MF),另取10只正常大鼠做正常对照组(Normal,N).血糖波动糖尿病组错时腹腔注射葡萄糖0.38 g·kg-1和皮下注射胰岛素1U制备糖尿病血糖波动模型.血糖波动6周后,进行水迷宫定位航行实验和空间探索实验,同时检测海马组织中Bax、Bcl-2的表达.[结果]M组、MS组、MF组比N组逃避潜伏期明显增长(P<0.05),空间探索实验结果显示60s内穿越平台所在位置的次数明显下降,在平台所在象限内的游泳距离明显减少(P<0.05).MF组和M组、MS组相比.逃避潜伏期进一步增长(P<0.05),空间探索实验结果显示60s内穿越平台所在位置次数也明显下降,在平台所在象限游泳距离也减少(P<0.05).MF组、M纽、MS组Bcl-2的表达较N组显著下降,Bax显著升高(P<0.05).MF组Bcl-2的表达较M组、MS组进一步下降,Bax进一步升高(P<0 05).[结论]血糖波动能加重损伤糖尿病大鼠的学习记忆功能,并促进海马组织的凋亡相关蛋白Bax/Bcl-2的表达,诱导海马神经元的凋亡.%[Objective] To explore the effection of glucose fluctuation on the cognitive function and the Bax/Bcl-2 in hippocampal of rats with type 2 diabetes mellitus. [Methods] The SD rats were randomly divided into four groups: normal group(N); diabetes model group(M);high glucose sustain diabetes model group(MS); glucose fluctuate diabetes model group(MF). The diabetic rats model was developed by injection with streptozotocin(STZ) 30 mg·kg-1, regularly staggered insulin 1U and glucose 0.38 mg·kg-1 administration was used to set up blood

  8. TIP30 regulates apoptosis-related genes in its apoptotic signal transduction pathway

    Institute of Scientific and Technical Information of China (English)

    Mei Shi; Xia Zhang; Ping Wang; Hong-Wei Zhang; Bai-He Zhang; Meng-Chao Wu

    2005-01-01

    AIM: To investigate the role of TIP30 in apoptotic signal pathway in hepatoblastoma cells and to provide a basis for TIP30 as a gene therapy candidate in the regression of hepatoblastoma cells.METHODS: Apoptosis of human hepatoblastoma cell lines HepG2 (p53 wild), Hep3B (p53 null) and PLC/RPF/5 (p53mutant) infected with Ad-TIP30 (bearing a wild type human Tip30 gene) were analyzed and p53, Bax and Bclxl expression levels were compared among these cells.MlT assay, DNA fragmentation, in situ 3' end labeling of DNA, annexin-Ⅴ FITC staining were used to detect cell death and apoptosis in cells at various time intervals subsequent to infection, and to determine whether TIP30 had an effect on the expression levels of some apoptosis-related gene products such as Bax, p53 and Bcl-xl. A similar time course experiment was performed by Western blotting.RESULTS: In MTT assay, the viability of HepG2 cells decreased significantly from 99.7% to 10% and displayed more massive cell death within 5-8 d than Hep3B and PLC/RPF/5 cells, with their viability decreased from 97.8% to 44.3% and 98.1% to 50.4%, respectively. In annexin-ⅤFITC assay, the percentage of apoptosis cells in HepG2cells was two to three-fold higher than that in control cells (infected with Ad-GFP), two-fold higher than that in Hep3B cells and 1.4-fold higher than that in PLC/RPF/5 cells 36 h after infection, respectively. Moreover, in HepG2 cells, the p53 began to increase 6-8 h after infection, reaching a maximum level between 8 and 12 h after infection and then dropped. Bax showed a similar increase in the cells as p53 reached the maximum at 8-12 h and subsequently decreased. Interestingly, Bcl-xl protein levels were down regulated during 24 to 36 h after Ad-TIP30 infection. In contrast, ectopic expression of TIP30 in Hep3B and PLC/RPF/5 cells had no effect on the regulation of Bax expression, but had an effect on Bcl-xl levels. In comparison with HepG2 cells, these data suggested that up-regulation of p53

  9. 氯胺酮对兔膝骨性关节炎软骨细胞Bcl-2及Bax表达的影响%Influence of Ketamine (KET,Ketamine) of intra-articular injection of rabbit chondrocytes and Bax expression

    Institute of Scientific and Technical Information of China (English)

    王雯; 王林; 陆巍

    2015-01-01

    Objective To explore the different concentrations of after the Bcl‐2 ,and the influence of Bax expres‐sion .Method The rest of rabbits with left knee immobilized in extension position to establish osteoarthritis models , then ,divided the models into model group ,ketamine60 group ,ketamine100 group and ketamine200 group in ran‐dom .They were received 0 .4 ml physiologic saline ,KET of 60μmol/L ,KET of 100μmol/L ,KET of 200μmol/L intra‐articular respectively ,two times a week for 4 weeks ,and sacrificed one week after last injection .To return af‐ter knee joint cartilage specimens corresponding processing ,using immunohistochemical method to observe the Bcl‐2 in the articular cartilage and the expression of bax .Result Compared with model group ,the injection of ketamine and promote cartilage cells apoptosis gene Bcl‐2 and Bax genes are changed (P<0 .05) .ketamine200 group of antiapop‐totic effect is the most significant (P<0 .05) .Conclusion Ketamine can suppress the cartilage cell apoptosis induced by osteoarthritis .%目的:观察不同浓度氯胺酮(Ketamine ,KET )家兔关节腔内注射后软骨细胞Bcl‐2及Bax表达的影响。方法通过管型石膏伸直位制动法造模成功后,随机分为模型组、氯胺酮60组、氯胺酮100组、氯胺酮200组,其中模型组、氯胺酮60组、氯胺酮100组、氯胺酮200组分别给予0.4 mL的生理盐水、KET(60μmol/L)、KET(100μmol/L)、KET(200μmol/L)关节腔内注射,每周2次,共进行4周,最后一次注射后1周处死动物。留取膝关节软骨标本作相应处理后,利用免疫组化法观察关节软骨中Bcl‐2及Bax的表达情况。结果与模型组比较,注射氯胺酮的软骨细胞促凋亡基因Bcl‐2及Bax均改变(P<0.05),其中氯胺酮200组的抗凋亡效果最为显著(P<0.05)。结论氯胺酮可以抑制骨关节炎诱导的软骨细胞凋亡。

  10. Anti-apoptotic mechanism of Bacoside rich extract against reactive nitrogen species induced activation of iNOS/Bax/caspase 3 mediated apoptosis in L132 cell line.

    Science.gov (United States)

    Anand, T; Pandareesh, M D; Bhat, Pratiksha V; Venkataramana, M

    2014-10-01

    Nitric oxide is a highly reactive free radical gas that reacts with a wide range of bio-molecules to produce reactive nitrogen species and exerts nitrative stress. Bacopa monniera is a traditional folk and ayurvedic medicine known to alleviate a variety of disorders. Aim of the present study is to evaluate the protective propensity of Bacopa monniera extract (BME) through its oxido-nitrosative and anti-apoptotic mechanism to attenuate sodium nitroprusside (SNP)-induced apoptosis in a human embryonic lung epithelial cell line (L132). Our results elucidate that pre-treatment of L132 cells with BME ameliorates the mitochondrial and plasma membrane damage induced by SNP as evidenced by MTT and LDH leakage assays. BME pre-treatment inhibited NO generation by down-regulating inducible nitric oxide synthase expression. BME exhibited potent antioxidant activity by up-regulating the antioxidant enzymes. SNP-induced damage to cellular, nuclear and mitochondrial integrity was also restored by BME, which was confirmed by ROS estimation, comet assay and mitochondrial membrane potential assays respectively. BME pre-treatment efficiently attenuated the SNP-induced apoptotic biomarkers such as Bax, cytochrome-c and caspase-3, which orchestrate the proteolytic damage of the cell. By considering all these findings, we report that BME protects L132 cells against SNP-induced toxicity via its free radical scavenging and anti-apoptotic mechanism.

  11. Enhancement of Photon Absorption on BaxSr1-xTiO3 Thin-Film Semiconductor Using Photonic Crystal

    Directory of Open Access Journals (Sweden)

    Abd. Wahidin Nuayi

    2014-01-01

    Full Text Available Enhancement of photon absorption on barium strontium titanate (BaxSr1-xTiO3 thin-film semiconductor for mole fraction x=0.25, 0.35, 0.45, and 0.55 using one-dimensional photonic crystal with defect was investigated experimentally. The thin film was grown on transparent conductive oxide (TCO substrate using chemical solution deposition method and annealed at 500°C for 15 hours with increasing rate of 1.6°C/min. From optical characterization in visible spectrum it was found that the average absorption percentages are 92.04%, 83.55%, 91.16%, and 80.12%, respectively. The BST thin film with embedded photonic crystal exhibited a relatively significant enhancement on photon absorption, with increasing value of 3.96%, 7.07%, 3.04%, and 13.33% for the respective mole fraction and demonstrating absorbance characteristic with flat feature. In addition, we also discuss the thin-film properties of attenuation constant and electrical conductivity.

  12. Study on the in-plane electrical resistivity and thermoelectric power in single crystals of La2-xBaxCuO4

    Institute of Scientific and Technical Information of China (English)

    李鹏程; 杨宏顺; 李志权; 柴一晟; 曹烈兆

    2002-01-01

    The in-plane electrical resistivity and thermoelectric power have been measured on single crystals ofLa2-xBaxCuO4 at around x=0.125. The room temperature resistivity and thermopower have their maximum val-ues at x=0.125, indicating that the carrier concentration is the minimum and the carriers are most strongly localized atx=0.125. The observed semiconductor-like behaviour can be well described by the weak-localized quasi-two-dimensionalstate. The steep rise in electric resistivity of the sample at x=0.125 below 70K is attributed to the formation of staticstripe-order of holes and spins, which are pinned by the low-temperature tetragonal (LTT) structure, as discovered inLa1.4sNd0.4Sr0.12CuO4.The temperature dependence of electric resistivity below 70K is still well described by theformula p ∝ lnT. A definite change in the slope of thermopower is observed at the low-temperature orthorhombic-LTTstructural phase transition temperature. The origin of the 1/8 anomaly is discussed in the text.

  13. Bcl-2、Bax及M30在Bowen病中的表达%Expression of Bcl-2、 Bax and M30 in Bowen's disease

    Institute of Scientific and Technical Information of China (English)

    张敏; 张谊之; 王琳; 李俸媛

    2002-01-01

    为了研究细胞凋亡与Bowen病发病的关系,我们采用免疫组化方法检测了28例Bowen病皮损中Bcl-2、Bax及M30的表达.结果发现M30及Bax染色阳性分别见于4例和8例表皮角质形成细胞,Bcl-2在10例表皮角质形成细胞和12例真皮淋巴细胞呈阳性表达;Bcl-2表达阳性率高于Bax,但无统计学差异(χ2=2.1053,P>0.05);M30与Bax表达显著正相关(r=0.6455,P0.05).提示Bowen病发病可能与表皮角质形成细胞凋亡降低有关.

  14. Phenomenological theory of phase transitions in epitaxial BaxSr1-xTiO3 thin films on (111)-oriented cubic substrates

    Science.gov (United States)

    Shirokov, V. B.; Shakhovoy, R. A.; Razumnaya, A. G.; Yuzyuk, Yu. I.

    2015-07-01

    A phenomenological thermodynamic theory of BaxSr1-xTiO3 (BST-x) thin films epitaxially grown on (111)-oriented cubic substrates is developed using the Landau-Devonshire approach. The group-theoretical analysis of the low-symmetry phases was performed taking into account two order parameters: the polarization related to ionic shifts in polar zone-center F1u mode and the out-of-phase rotation of TiO6 octahedra corresponding to the R25 zone-boundary mode in the parent cubic phase P m 3 ¯ m . The eight-order thermodynamic potential for BST-x solid solutions was developed and analyzed. We constructed the "concentration-misfit strain" phase diagram for BST-x thin films at room temperature and found that polar rhombohedral R3m phase with the polarization normal to the substrate is stable for x > 0.72 and negative misfit strains, while ferroelectric monoclinic C2 and Cm phases with in-plane polarization are stable for much smaller x and positive or slightly negative misfit strains. We constructed the "temperature-misfit strain" phase diagrams for several concentrations (x = 1, 0.8, 0.6, 0.4, and 0.2). Systematic changes of the phase transition lines between the paraelectric and ferroelectric phases are discussed. The phase diagrams are useful for practical applications in thin-film engineering.

  15. Moessbauer study of the relationship between magnetic properties and Jahn-Teller distortion in La1-xBax(Mn,Fe)O3 perovskites

    International Nuclear Information System (INIS)

    The local structural distortion and its effect on the magnetic and magnetotransport properties of the Fe-doped La1-xBaxMnO3 (0.00≤x≤0.35) compounds were investigated by means of X-ray diffraction, Moessbauer spectroscopy, and magnetization measurements. The Moessbauer spectra show clear evidence of the local structural distortion of the Mn(Fe)O6 octahedron on the basis of non-zero nuclear quadrupole interactions for high-spin Fe3+ ions. It was found that the local structural distortion decreases significantly with the introduction of a few per cent of Ba, reaches a minimum around x=0.13, and then increases slightly on further increasing the Ba concentration. The Ba-concentration dependence of the Jahn-Teller coupling strength estimated from the Moessbauer results was found to be consistent with the magnetic properties. Consequently, the connection between this local structural distortion and the colossal magnetoresistive behavior of this family of materials can be well explained. (copyright 2005 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  16. Layered Bi4BaxV2-xO11-(3x/2)-δ perovskite oxide as solid electrolyte for intermediate temperature solid oxide fuel cells

    International Nuclear Information System (INIS)

    A continuous series of Bi4BaxV2-xO11-(3x/2)-δ solid solutions are prepared by conventional solid state reactions. The polymorphism and electrical properties of these samples are studied by FT-IR spectroscopy, X-ray diffraction, differential thermal analysis (DTA) and AC impedance. The solid solutions with composition 0.07≤x≤0.13 are isostructural with the monoclinic phase α-Bi4V2O11. However, orthorhombic β-phase is observed for x=0.17 and tetragonal γ-phase is stabilized for x≤0.30. X-ray and DTA results reveal the occurrence of α↔γ transition for x≤0.13, β↔γ transition for x=0.17 and γ'→γ transition for x≥0.20. AC impedance plots at 280 deg. C for all compositions show greater contribution of grain to ionic conductivity than grain boundary. The highest ionic conductivity, σ300oC=4.456x10-5 S cm-1 is observed for x=0.17. The ionic conductivity of the substituted compounds is higher than the parent α-Bi4V2O11, due to the increased oxygen ion vacancies generated as a result of barium doping.

  17. Protective Effect of Aliskiren in Experimental Ischemic Stroke: Up-Regulated p-PI3K, p-AKT, Bcl-2 Expression, Attenuated Bax Expression.

    Science.gov (United States)

    Miao, Jiangyong; Wang, Lina; Zhang, Xiangjian; Zhu, Chunhua; Cui, Lili; Ji, Hui; Liu, Ying; Wang, Xiaolu

    2016-09-01

    Aliskiren (ALK), a pharmacological renin inhibitor, is an effective antihypertensive drug and has potent anti-apoptotic activity, but it is currently unknown whether ALK is able to attenuate brain damage caused by acute cerebral ischemia independent of its blood pressure-lowering effects. This study aimed to investigate the role of ALK and its potential mechanism in cerebral ischemia. C57/BL6 mice were subjected to transient middle cerebral artery occlusion (tMCAO) and treated for 5 days with Vehicle or ALK (10 or 25 mg/kg per day via intragastric administration), whereas Sham-operated animals served as controls. Treatment with ALK significantly improved neurological deficits, infarct volume, brain water content and Nissl bodies after stroke (P < 0.05), which did not affect systemic blood pressure. Furthermore, the protection of ALK was also related to decreased levels of apoptosis in mice by enhanced activation of phosphatidylinositol 3-kinase (PI3K)/AKT pathway, increased level of Bcl-2 and reduced Bax expression (P < 0.05). In addition, ALK's effects were reversed by PI3K inhibitors LY294002 (P < 0.05). Our data indicated that ALK protected the brain from reperfusion injuries without affecting blood pressure, and this effect may be through PI3K/AKT signaling pathway. PMID:27180190

  18. Resistance Fluctuation Spectroscopy of Charge Stripes and Intertwined Orders in the Phase Diagram of La2-xBaxCuO4

    Science.gov (United States)

    Weis, Adam; Fizari, Mounir; Hamilton, David; Wells, Azton; Lane, Justin; Chung, So Ra; Sellappan, Pathikumar; Kriven, Waltraud; van Harlingen, Dale

    The unusual phase diagram of La2-xBaxCuO4 (LBCO) near x=1/8 doping suggests a complex intertwined relationship between high-temperature superconductivity, charge stripes, spin order, and phase coherence. The charge stripe state's short-range conductance anisotropy may be observable as fluctuations in resistance. In thin film LBCO devices grown by pulsed laser deposition, our time-resolved resistance measurements have revealed an onset of resistance noise at dopings and critical temperatures consistent with charge stripes. The phase diagram of LBCO is explored by comparing the noise onset signature of charge order to measurements of superconductivity, the Hall effect, and other phenomena. I will briefly discuss the relevance of our results in LBCO thin films and crystals to a proposed ''pair-density-wave'' state near x=1/8. This research was supported by the DOE-BES under Grant DE-SC0012368, through the Frederick Seitz Materials Research Laboratory at the University of Illinois at Urbana-Champaign. SRC was sponsored by NSF-REU 13-59126.

  19. Effect of Buspirone, Fluoxetine and 8-OH-DPAT on Striatal Expression of Bax, Caspase-3 and Bcl-2 Proteins in 6-Hydroxydopamine-Induced Hemi-Parkinsonian Rats

    OpenAIRE

    Hamdollah Sharifi; Alireza Nayebi; Safar Farajnia; Rasool Haddadi

    2015-01-01

    Purpose: The exact pathogenesis of sporadic parkinson’s disease (PD) is still unclear. Numerous evidences suggest involvement of apoptosis in the death of dopaminergic neurons. In this study we investigated the effect of sub-chronic administration of buspirone, fluoxetine and 8-hydroxy-2-[di-n-propylamino]tetralin (8-OH-DPAT) in 6-hydroxydopamine (6-OHDA)-lesioned rats and assayed striatal concentrations of apoptotic (Bax, Caspase3) and anti-apoptotic (Bcl-2) proteins. M...

  20. Der Einfluss der Anästhetika Sevofluran und Propofol auf die Regulation der apoptoseassoziierten Proteine Bax, Bcl-2, Mdm-2 und p53 nach inkompletter zerebraler Hemisphärenischämie bei der Ratte

    OpenAIRE

    Bachl, Monika Maria

    2005-01-01

    Der Einfluss der Anästhetika Sevofluran und Propofol auf apoptoseassoziierte Proteine während zerebraler Ischämie ist bisher nicht erforscht. In der vorliegenden Studie wurden die Effekte dieser Narkotika auf die Regulation der Apoptosefaktoren Bax, Bcl-2, Mdm-2 und p53 bei 36 narkotisierten Sprague-Dawley-Ratten untersucht, bei denen eine inkomplette zerebrale Hemisphärenischämie mit anschließender Reperfusion induziert wurde. Die Apoptosefaktoren wurden mittels Immunfluoreszenz- und Western...

  1. P53,Bax,Bcl-2蛋白表达及细胞凋亡在急性放射性皮肤溃疡发生发展过程中的作用探讨%The role of P53, Bax, Bcl-2 expression and cell apoptosis in the formation and development of acute radiation-induced skin ulcers

    Institute of Scientific and Technical Information of China (English)

    谷庆阳; 曹卫红; 王德文; 高亚兵; 杨志祥; 赵坡

    2001-01-01

    目的:研究细胞凋亡及一些凋亡相关基因(p53,bcl-2,bax)的表达在急性放射性皮肤溃疡发生发展过程中的作用.方法:采用Wistar大鼠以60Co γ射线进行局部照射,建立急性放射性皮肤溃疡动物模型,观察病变40 d,然后采用免疫组化方法检测皮肤溃疡组织中P53,Bcl-2,Bax蛋白表达,并采用原位末端标记法(TUNEL)检测细胞凋亡.结果:照后14 d照射野内开始出现皮肤溃疡,之后逐渐扩大、融合、加深;照后11~40 d,P53蛋白表达明显增强,主要定位于血管内皮细胞和小血管平滑肌中;照后14~21 d为Bax蛋白表达高峰,之后逐渐减弱,主要定位于血管内皮细胞、部分成纤维细胞及新生表皮细胞中;Bcl-2则在照后1~11 d呈弱或中度阳性,定位于表皮、毛囊上皮及血管内皮中,之后为阴性或可疑阳性;照后11~35 d,上述细胞特别是血管内皮细胞凋亡率较正常伤口愈合早期增高.结论:辐射诱导的P53,Bax,Bcl-2表达的变化及细胞凋亡率特别是血管内皮细胞凋亡率的增高与放射性皮肤溃疡发生、发展及难愈合(不能形成有效肉芽组织)的分子机制相关.%Objective:To study the expression of P53, Bax, Bcl-2 proteins and the role of cell apoptosis in the formation and development of acute radiation-induced skin ulcers.Methods:A rat model which was locally irradiated with 60 Co γ-rays was used, and the pathological changes were observed for 40 days. Immunohistochemistry and TUNEL assay were performed which enabled the detection of P53, Bax, Bcl-2 and cell apoptosis during the formation and development of radiation skin ulcers.Results: Skin ulcers were found on day 14 after irradiation, and enlarged and deepened gradually during the observation period. P53 was over expressed during days 11 to 40 after irradiation and was localized in vascular endotheliocytes and smooth muscle cells. Bax was moderately positive during days 14 to 21 and weakly positive during days

  2. Cisplatin-induced apoptosis in non-small-cell lung cancer cells is dependent on Bax- and Bak-induction pathway and synergistically activated by BH3-mimetic ABT-263 in p53 wild-type and mutant cells.

    Science.gov (United States)

    Matsumoto, Masaru; Nakajima, Wataru; Seike, Masahiro; Gemma, Akihiko; Tanaka, Nobuyuki

    2016-04-29

    Cisplatin is a highly effective anticancer drug for treatment of various tumors including non-small-cell lung cancer (NSCLC), and is especially useful in cases nonresponsive to molecular-targeted drugs. Accumulating evidence has shown that cisplatin activates the p53-dependent apoptotic pathway, but it also induces apoptosis in p53-mutated cancer cells. Here we demonstrated that DNA-damage inducible proapoptotic BH3 (Bcl-2 homology region 3)-only Bcl-2 family members, Noxa, Puma, Bim and Bid, are not involved in cisplatin-induced apoptosis in human NSCLC cell lines. In contrast, the expression of proapoptotic multidomain Bcl-2-family members, Bak and Bax, was induced by cisplatin in p53-dependent and -independent manners, respectively. Moreover, in wild-type p53-expressing cells, cisplatin mainly used the Bak-dependent apoptotic pathway, but this apoptotic pathway shifted to the Bax-dependent pathway by loss-of-function of p53. Furthermore, both Bak- and Bax-induced apoptosis was enhanced by the antiapoptotic Bcl-2 family member, Bcl-XL knockdown, but not by Mcl-1 knockdown. From this result, we tested the effect of ABT-263 (Navitoclax), the specific inhibitor of Bcl-2 and Bcl-XL, but not Mcl-1, and found that ABT-263 synergistically enhanced cisplatin-induced apoptosis in NSCLC cells in the presence or absence of p53. These results indicate a novel regulatory system in cisplatin-induced NSCLC cell apoptosis, and a candidate efficient combination chemotherapy method against lung cancers.

  3. Combination of cold atmospheric plasma and iron nanoparticles in breast cancer: gene expression and apoptosis study

    Directory of Open Access Journals (Sweden)

    Jalili A

    2016-09-01

    Full Text Available Azam Jalili,1 Shiva Irani,1 Reza Mirfakhraie2 1Department of Biology, Science and Research Branch, Islamic Azad University, 2Department of Medical Genetics, Shahid Beheshti University of Medical Sciences, Tehran, Iran Background: Current cancer treatments have unexpected side effects of which the death of normal cells is one. In some cancers, iron nanoparticles (NPs can be subjected to diagnosis and passive targeting treatment. Cold atmospheric plasma (CAP has a proven induction of selective cell death ability. In this study, we have attempted to analyze the synergy between CAP and iron NPs in human breast adenocarcinoma cells (MCF-7.Materials and methods: In vitro cytotoxicity of CAP treatment and NPs in cells measured by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT assay and cell death was shown by 4',6-diamidino-2-phenylindole and annexin V staining. Fluctuations in BAX and BCL-2 gene expression were investigated by means of real-time polymerase chain reaction.Results: MTT assay results showed that combination of plasma and iron NPs decreased the viability of cancer cells significantly (P<0.05. Real-time analysis showed that the combination therapy induced shifting the BAX/BCL-2 ratio in favor of apoptosis.Conclusion: Our data indicate that synergy between CAP and iron NPs can be applied in breast cancer treatment selectively. Keywords: breast cancer, cold atmospheric plasma, iron nanoparticles, BAX, BCL-2

  4. THE EXPRESSIONS OF HIF-1α AND Bax AND THEIR CORRELATION IN SUDDEN CARDIAC DEATH%心脏性猝死心肌组织HIF-1α和Bax表达及其相关性

    Institute of Scientific and Technical Information of China (English)

    刘德衍; 袁磊; 杨彦华; 纪萍; 于建宪; 张七一; 林梅

    2011-01-01

    目的 探讨缺氧诱导因子1 alpha(HIF-1α)和Bax蛋白在心脏性猝死(SCD)者心肌组织中的表达与相关性,并为法医学鉴定SCD提供客观依据.方法 应用免疫组织化学方法,检测25例SCD者心肌组织中HIF-1α和Bax蛋白的表达,并以25例非SCD者心肌组织作为对照.结果 SCD者心肌组织HIF-1α与Bax的表达明显高于对照组(uc=6.083、5.573,P<0.01).在SCD心肌组织中,HIF-1α表达与Bax表达呈正相关(r=0.736,P<0.01).结论 HIF-1α与Bax表达的增高与SCD的发生有一定的相关性,HIF-1α与Bax可望作为诊断SCD较为客观的病理形态学指标.%Objective To investigate the expressions of hypoxia-inducible factor-1 alpha (HIF-1α) and Bcl-2 associated X protein (Bax) and their correlation in myocardium of sudden cardiac death (SCD) and provide an objective evidence for forensic identification.Methods The expressions of HIF-la and Bax in myocardium of 25 SCD were detected using immunohistochemical technique, and 25 of non-sudden cardiac death served as controls.Results The expressions of HIF-1α and Bax in myocardium of the SCD were significantly higher than that of the control (Uc = 6.083,5.573;P<0.01), and the expressions were positively correlated (r=0.736,P<0.01) Conclusion The elevation of the expressions of HIF-1α and Bax is associated with SCD to a certain extent.The HIF-1α and Bax will hopefully become a relatively objective pathomorphologic index in the diagnosis of SCD.

  5. MicroRNA-650 was a prognostic factor in human lung adenocarcinoma and confers the docetaxel chemoresistance of lung adenocarcinoma cells via regulating Bcl-2/Bax expression.

    Directory of Open Access Journals (Sweden)

    Jia-Yuan Huang

    Full Text Available Increasing evidence shows that dysregulation of microRNAs (miRNAs is involved in malignant transformation. We investigated the clinical significance of miR-650 and its involvement in chemoresistance to docetaxel. Our results showed that the relative expression level of miR-650 was significantly higher in LAD tissues than in corresponding nontumor tissues and high level of miR-650 expression was found to be significantly associated with high incidence of lymph node metastasis, advanced clinical stage and poor prognosis of LAD patients. Univariate and multivariate analyses indicated that high miR-650 expression was an independent prognostic factor for survival. Also, we found that the level of miR-650 in LAD tissues was correlated with the response of patients to docetaxel-based chemotherapy. Silencing of miR-650 could increase the in vitro sensitivity of docetaxel-resistant LAD cells to docetaxel, while upregulation of miR-650 decreased the sensitivity of parental LAD cells to docetaxel both in vitro and in vivo. Additionally, silencing of miR-650 could enhance the caspase-3-dependent apoptosis, which might be correlated with the decreased ratio of Bcl-2/Bax. Further researches suggested that inhibitor of growth 4 (ING4 was a direct target of miR-650. Downregulated or upregulated ING4 expression could partially rescue the effects of miR-650 inhibitor or mimics in docetaxel-resistant or parental LAD cells. Furthermore, we found that ING4 was upregulated in docetaxel-responding LAD tissues, and its expression was inversely correlated with miR-650. Thus, miR-650 is a novel prognostic marker in LAD and its expression is a potential indicator of chemosensitivity to docetaxel-based chemotherapy regimen.

  6. The influence of sleep deprivation on expression of apoptosis regulatory proteins p53, bcl-2 and bax following rat tongue carcinogenesis induced by 4-nitroquinoline 1-oxide

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    Juliana Noguti

    2013-01-01

    Full Text Available Background: The aim of this study was to evaluate whether paradoxical sleep deprivation could affects the mechanisms and pathways essentials for cancer cells in tongue cancer induced by 4-nitroquinole 1-oxide in Wistar rats. Materials and Methods: For this purpose, the animals were distributed into 4 groups of 5 animals each treated with 50 ppm 4 nitroquinoline 1 oxide (4 NQO solution through their drinking water for 4 and 12 weeks. The animals were submitted to paradoxical sleep deprivation (PSD for 72 h using the modified multiple platform method, which consisted of placing 5 mice in a cage (41 × 34 × 16 cm containing 10 circular platforms (3.5 cm in diameter with water 1 cm below the upper surface. The investigations were conducted using immunohistochemistry of p53, Bax and Bcl-2 proteins related to apoptosis and its pathways. Statistical analysis was performed by Kruskal-Wallis non-parametric test followed by the Dunn′s test using SPSS software pack (version 1.0. P value < 0.05 was considered for statistic significance. Results: Although no histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure in all groups, in 12 weeks were observed pre-neoplasic lesions. Data analysis revealed statistically significant differences ( P < 0.05 in 4 weeks group for p53 and for bcl-2 and for all immunomarkers after 12 weeks of 4NQO administration. Conclusion: Our results reveal that sleep deprivation exerted alterations in proteins associated with proliferation and apoptosis in carcinogenesis.

  7. Tubeimoside-1 induces glioma apoptosis through regulation of Bax/Bcl-2 and the ROS/Cytochrome C/Caspase-3 pathway

    Directory of Open Access Journals (Sweden)

    Jia G

    2015-01-01

    Full Text Available Geng Jia,1,* Qiang Wang,2,* Rong Wang,2,* Danni Deng,2 Lian Xue,2 Naiyuan Shao,1 Yi Zhang,1 Xiwei Xia,1 Feng Zhi,2 Yilin Yang1,2 1Department of Neurosurgery, Third Affiliated Hospital of Soochow University, Jiangsu, People’s Republic of China; 2Modern Medical Research Center, Third Affiliated Hospital of Soochow University, Jiangsu, People’s Republic of China * These authors contributed equally to this workBackground: Tubeimoside-1 (TBMS1 is a natural compound isolated from tubeimoside, which has been widely used as a traditional Chinese herbal medicine. The purpose of the present study is to investigate the anti-tumor effect and the underling mechanism of TBMS1 on glioma cancer cells.Methods: The MTT assay was performed to evaluate the effect of TBMS1 on glioma cell proliferation. The fluorescent microscopy and flow cytometry analysis were performed to evaluate the effect of TBMS1 on glioma cell apoptosis. The Western blot analysis was used to evaluate the protein change.Results: TBMS1 inhibited glioma cancer cell proliferation in a dose- and time-dependent manner. Fluorescent microscopy and flow cytometry analysis demonstrated that TBMS1 induced glioma cell apoptosis in a concentration-dependent manner. Western blotting showed that TBMS1 induced apoptosis by increasing the expression of Bax and downregulating the level of Bcl-2. Furthermore, we found that TBMS1 induced apoptosis by increasing the concentration of reactive oxygen species through the release of Cytochrome C and activation of Caspase-3.Conclusion: These findings indicate that TBMS1 may be developed as a possible therapeutic agent for the management of glioma. Keywords: Tubeimoside-1, glioma, proliferation, apoptosis

  8. In situ hybridisation detects pro-apoptotic gene expression of a Bcl-2 family member in white syndrome-affected coral.

    Science.gov (United States)

    Ainsworth, T D; Knack, B; Ukani, L; Seneca, F; Weiss, Y; Leggat, W

    2015-12-01

    White syndrome has been described as one of the most prolific diseases on the Great Barrier Reef. Previously, apoptotic cell death has been described as the mechanism driving the characteristic rapid tissue loss associated with this disease, but the molecular mechanisms controlling apoptotic cell death in coral disease have yet to be investigated. In situ methods were used to study the expression patterns of 2 distinct regulators of apoptosis in Acropora hyacinthus tissues undergoing white syndrome and apoptotic cell death. Apoptotic genes within the Bcl-2 family were not localized in apparently healthy coral tissues. However, a Bcl-2 family member (bax-like) was found to localize to cells and tissues affected by white syndrome and those with morphological evidence for apoptosis. A potential up-regulation of pro-apoptotic or bax-like gene expression in tissues with apoptotic cell death adjacent to disease lesions is consistent with apoptosis being the primary cause of rapid tissue loss in coral affected by white syndrome. Pro-apoptotic (bax-like) expression in desmocytes and the basal tissue layer, the calicodermis, distant from the disease lesion suggests that apoptosis may also underlie the sloughing of healthy tissues associated with the characteristic, rapid spread of tissue loss, evident of this disease. This study also shows that in situ hybridisation is an effective tool for studying gene expression in adult corals, and wider application of these methods should allow a better understanding of many aspects of coral biology and disease pathology. PMID:26648107

  9. Overexpression of the hydatidiform mole-related gene F10 inhibits apoptosis in A549 cells through downregulation of BCL2-associated X protein and caspase-3.

    Science.gov (United States)

    Song, Yali; Zhang, Gong; Zhu, Xiulan; Pang, Zhanjun; Xing, Fuqi; Quan, Song

    2012-09-01

    The aim of this study was to investigate how the overexpression of the hydatidiform mole-related gene F10 affects apoptosis in human lung cancer A549 cells. A549 cells were transfected with pEGFP-N1-F10 (A549-F10) or pEGFP-N1 empty vector (A549-empty). Untransfected A549, A549-F10 or A549-empty cells were examined using the MTT cell proliferation assay and the TUNEL-FITC/Hoechst 33258 apoptosis assay. Western blotting was used to examine the expression levels of the pro-apoptotic genes, BCL2-associated X protein (BAX) and caspase-3. F10 was stably expressed in A549 cells. From 12 h, A549-F10 cells proliferated markedly faster than the untransfected and A549-empty cells. F10 overexpression also significantly inhibited apoptosis, as shown by the reduced number of TUNEL and Hoechst 33258 double-positive cells. This inhibition was likely due to an F10-induced reduction in the BAX and caspase-3 levels. The results of this study indicate that F10 overexpression inhibits apoptosis in A549 cells through the downregulation of the pro-apoptotic genes BAX and caspase-3. PMID:23741243

  10. Effect of Shenfu parenteral injection on the expressions of Bcl-2, Bax and c-Fos proteins in ischemia reperfusion myocardium of rats%参附注射液影响大鼠缺血再灌注心肌Bcl-2,Bax与c-Fos蛋白的表达

    Institute of Scientific and Technical Information of China (English)

    陈玉培; 牟崇明; 季道如; 但伶; 龚文婷; 王莉莎

    2006-01-01

    -2/Bax比率显著升高(P<0.01).结论:参附注射液对缺血再灌注心肌保护效应可能与其促进Bcl-2蛋白高表达、抑制Bax与c-Fos蛋白表达、增加Bcl-2/Bax比率,从而抑制心肌细胞凋亡有关.%BACKGROUND: It has been confirmed that Shenfu parenteral injection can ameliorate and treat various shocks, heart failure, myocardial ischemia and supraventricular/ventricular arrhythmia, and it also has a good protective effect on myocardial ischemia/reperfusion injury in rats.OBJECTIVE: To observe the effects of Shenfu parenteral injection on the protein expressions of myocardial apoptosis-related genes of Bcl-2, Bax and c-Fos in rats with acute ischemia/reperfusion injury.DESIGN: A complete randomized grouping design, controlled experiment.SETTING: Department of Anesthesiology, the Second Affiliated Hospital,Chongqing University of Medical Sciences.MATERIALS: The experiments were carried out in the Staff Room of Anesthesiology, the Second Affiliated Hospital, Chongqing University of Medical Sciences from April to December in 2004. Thirty-five healthy adult Wistar rats were provided by the experimental animaI center of Daping Hospital, Third Military Medical University of Chinese PLA. Shenfu parenteral injection was the TCM formula of Shenfu Tang, which is for recuperating depleted yang and rescuing the patient from collapse, and its main components are ginsenoside and aconitum alkaloid. It was the product of Yaan Sanjiu Pharmaceutical Co., Ltd., 10 mL/piece, the batch number was 030110.METHODS: In vivo models of myocardial ischemia/reperfusion injury were used. The 35 rats were divided into 5 groups according to the number of random number table, with 7 rats in each group: ① Sham-operated group: The rats were treated with only insertion of thread without ligation, followed by intravenous injection of saline (8 mL/kg), and then observed for 120 minutes. ② Shenfu parenteral injection 30-minute group: The rats were treated with intravenous

  11. 大骨节病关节软骨真菌毒素环境反应基因表达谱研究%Gene expression profiling of mycotoxin-related environmental response genes in the articular cartilage of Kashin-Beck disease

    Institute of Scientific and Technical Information of China (English)

    张峰; 王伟卓; 郭雄; 武世勋; 王立新

    2012-01-01

    Objective To compare the expression profile of mycotoxin-related environmental response genes (MERGs) in the articular cartilage of patients with Kashin-Beck disease (KBD) and healthy controls,and explore the relationship between MERG and KBD.Methods Articular cartilage specimens were collected from 9 healthy human subjects and 9 adult KBD patients.Agilent microarray was used to evaluate the expression levels of MERG in cartilage specimens,and the expression ratios of MERG between KBD and healthy controls were calculated.GSEA software was used to calculate the NES scores and P values of gene ontology(GO).Results ①T-2 toxin,deoxynivalenol,zearalenone,aflatoxin B1,fumonisin B1 and ochratoxin A related 15 MERGs presented expression differences between KBD and healthy controls(ratios > 2.0 or < 0.5).Thirteen MERGs were up-regulated in KBD,including BAX,BCL2,COL5A2,FER1L3,GSTT2,IGFBP2,IGFBP4,PDE8B,SOCS3,THBS1,TMSL8,VGLL3 and TUBB2A (ratio > 2.0).Two MERGs,POSTN and FABP4,were down-regulated in KBD (ratio < 0.5).The 15 MERGs were involved in various biological processes; such as collage synthesis,apoptosis,metabolism,growth & development and so on.②Mycotoxin related 4 apoptosis GOs and 5 growth & development related GOs were up-regulated in KBD compared to healthy controls(NES > 0),including ANTI_APOPTOSIS,REGULATION_OF_PROGRAMMED_CELL_DEATH,APOPTOSIS_GO,REGULATION_OF_APOPTOSIS,ORGAN_MORPHOGENESIS,ANATOMICAL_STRUCTURE_DEVELOPMENT,ORGAN_DEVELOPMENT,SYSTEM_DEVELOPMENT and REGULATION OF DEVELOPMENTAL_PROCESS (NES > 0 and P < 0.05).Conclusions There are multiple mycotoxins related environmental response genes presenting significant expression difference between KBD cartilage and normal cartilage.Mycotoxin can affect the expression of MERGs in KBD articular cartilage,which might lead to dysfunction of chondrocytes,and articular cartilage lesions.%目的 比较分析真菌毒素环境反应基因在大骨节病(Kashin-Beck disease,KBD)和正常关节软骨

  12. Deletion of growth hormone receptor gene but not visceral fat removal decreases expression of apoptosis-related genes in the kidney-potential mechanism of lifespan extension.

    Science.gov (United States)

    Gesing, Adam; Masternak, Michal M; Wang, Feiya; Karbownik-Lewinska, Malgorzata; Bartke, Andrzej

    2012-04-01

    Mice homozygous for the targeted disruption of the growth hormone (GH) receptor (Ghr) gene (GH receptor knockout; GHRKO; KO) are hypoinsulinemic, highly insulin sensitive, normoglycemic, and long-lived. Visceral fat removal (VFR) is a surgical intervention which improves insulin signaling in normal (N) mice and rats and extends longevity in rats. We have previously demonstrated decreased expression level of certain pro-apoptotic genes in skeletal muscles and suggested that this may contribute to the regulation of longevity in GHRKO mice. Alterations in apoptosis-related genes expression in the kidneys also may potentially lead to lifespan extension. In this context, we decided to examine the renal expression of the following genes: caspase-3, caspase-9, caspase-8, bax, bad, bcl-2, Smac/DIABLO, Apaf-1, p53, and cytochrome c1 (cyc1) in male GHRKO and N mice subjected to VFR or sham surgery, at approximately 6 months of age. The kidneys were collected 2 months after VFR. As a result, caspase-3, caspase-9, and bax expressions were decreased in KO mice as compared to N animals. Expressions of Smac/DIABLO, caspase-8, bcl-2, bad, and p53 did not differ between KOs and N mice. VFR did not change the expression of the examined genes in KO or N mice. In conclusion, endocrine abnormalities in GHRKO mice result in decreased expression of pro-apoptotic genes and VFR did not alter the examined genes expression in N and KO mice. These data are consistent with a model in which alterations of GH signaling and/or insulin sensitivity lead to increased lifespan mediated by decreased renal expression of pro-apoptotic genes. PMID:21431351

  13. Expression of Smac and Bax in chronic periodontal tissues and their roles in apoptosis%慢性牙周炎龈组织中Smac和Bax的表达及其对细胞凋亡的作用

    Institute of Scientific and Technical Information of China (English)

    冯利; 董跟喜; 孙晓玮

    2012-01-01

    AIM: To investigate the expression of Smac and Bax in gingival tissues of chronic periodontitispatients. METHODS; Gingival tissues from 27 periodontitis patients and 27 periodontally healthy subjects were obtained from extracted teeth. Light microscope and transmission electron microscope were used to observe the morphology and structure of apoptotic cells in gingival tissues. Mitochondria] transmembrane potential ( △ψm) was measured by flow cytometry. The expression of Smac and Bax was detected with immunohistochemistry in gingival tissues. RESULTS; Under light microscope, apoptotic cells could be observed. Morphological and structural changes could beseen in mitochondria under transmission electron microscope. △ψm was significantly lower in chronic periodontal tissues as compared with healthy gingiva (P <0.05). Smac and Bax expression was significantly stronger in gingival epithelial and connective tissues of chronic periodontitis patients (P < 0. 05). Smac expression was positively related to Bax expression (P<0.05) in chronic periodontal tissues. CONCLUSION: Apoptotic cells exist in gingival tissue of chronic periodontitis patients. Smac expression is positively related to Bax expression. Both Smac and Bax expression are significantly enhanced, indicating that they might play important roles in the destruction of periodontal tissues.%目的:通过检测Smac和Bax蛋白在慢性牙周炎龈组织中的表达,旨在探讨二者的相关性及其在细胞凋亡中与牙周炎发生、发展的关系.方法:慢性牙周炎测试组27人,健康对照组27人,利用光镜和透射电镜观察凋亡细胞形态结构,流式细胞仪测定线粒体膜电位.免疫组化检测Smac、Bax在龈组织不同区域的表达.结果:光镜观察慢性牙周炎龈组织中存在有凋亡细胞;电镜观察凋亡细胞中线粒体形态结构改变;慢性牙周炎龈组织线粒体膜电位降低,与健康组比较,两组间有显著性差异(P<0.05);慢性牙周

  14. Histological structure and expression of Bax protein in Pavo cristatus kidney%白孔雀肾脏的组织结构及Bax蛋白在肾脏中的表达

    Institute of Scientific and Technical Information of China (English)

    俞诗源; 王小勇; 吴勍

    2012-01-01

    To study Pavo cristatus kidney histological structure and the expression of relevant active substances, histological structure is obvserved by light microscopy and expression of Bax protein is examined by immunohistochemical methods. The results indicate that Pavo cristatus kidney consists of many nephrons, collecting ducts and small amounts of areolar tissue. The glomerular capillary is simpler, renal tubules arrange closely and the quantity is more. The proximal tubule consists of monolayer cuboidal epithelium cells, with brush border and deeper color. Bax protein immune response positive material is mainly distributed in proximal tubule epithelium. Bax protein may be involved in the cell apoptosis of Pavo cristatus kidney and plays an important regulation role in the development of bird kidney.%为了搞清白孔雀(Pavo cristatus)肾脏的结构特征和相关活性物质的表达问题,利用生物显微技术观察了白孔雀肾脏的组织结构,用免疫组织化学方法检测了Bax蛋白在肾组织中的表达.结果显示白孔雀肾脏主要由许多肾单位、集合管和少量结缔组织组成.白孔雀肾小球毛细血管网较简单,肾小管之间排列紧密,数量较多,近端小管由单层立方上皮细胞组成,细胞顶端有刷状缘,细胞着色较深;Bax蛋白免疫反应阳性物质主要分布在近端小管上皮细胞;Bax蛋白可能与白孔雀肾脏细胞的凋亡有关.

  15. 激活素A、Bax和XIAP在特发性胎儿生长受限患者的表达及意义%The Expression and Significance of Activin A, Bax and XIAP in Idiopathic Fetal Growth Restriction

    Institute of Scientific and Technical Information of China (English)

    崔世红; 张婷; 刘萍萍; 程国梅; 贾国占; 管秀娟; 韩笑

    2012-01-01

    Objective:To investigate the expression and significance of Activin A in maternal and cord serum idiopathic fetal growth restriction (IFGR), B-cell lymphoma associated x protein (Bax), and X-linked inhibitor of apoptosis protein (XIAP) in placenta of idiopathic fetal growth restriction (IFGR). Methods:From July 2010 to May 2011, 30 pregnant women with IFGR underwent cesarean section in our hospital were selected as experimental group, and 30 normal term pregnant women underwent cesarean section due to social factors were matched as control group. Activin A in maternal and umbilical cord serum were analyzed by enzyme-linked immunosorbent assay (ELISA), while Bax and XIAP in placenta were detected by immunohis-tochemical method. Results:The level of Activin A both in maternal and cord serum of the experimental group (102. 659 ±16.467, 57.752 ±13.498) ng/ml ,were significantly higher than those in control group (75. 927 ±10. 519,29. 870 ±5. 992) ng/ml(P<0.05). The expression of Bax in placental syncytiotropho-blast in experimental group (146. 513 ± 10. 611) was significantly increased as compared to the control group (113. 672 ±9. 631) (P <0. 05). The expression of XIAP in placental syncytiotrophoblast in experimental group (114. 562 ±5.167) was significantly decreased as compared to the control group (144. 430 ± 7. 311) (P <0. 05). The level of Activin A in maternal and cord serum was positively correlated with the expression of Bax in placenta in experimental group (p<0. 05), whilst it showed negative correlation with the expression of XIAP in placenta in experimental group (P<0. 05). Moreover, the expression of XIAP in syn-cytiotrophoblast was found negetively correlated with Bax (P<0. 05). Conclusions:The increased Activin A in maternal and cord serum, as well as the increased expression of Bax and decreased expression of XIAP in placenta, may play important roles in the pathogenisis of IFGR. The increased Activin A may lead to excessive apoptosis of

  16. THE ROLES OF bcl-2 GENE FAMILY IN THE PULMONARY ARTERY REMODELING OF HYPOXIA PULMONARY HYPERTENSION IN RATS

    Institute of Scientific and Technical Information of China (English)

    杨成; 王胜发; 梁桃; 王巨; 王凯; 王柏春

    2001-01-01

    Objective. To investigate the roles of apoptosis in the pulmonary artery remodeling of pulmonary hypertension secondary to hypoxia and illustrate the relative genes expression.Methods. Thirty rats were divided into hypoxia group(10%O2, 8h/d) and normal control group. On the 15th day of hypoxia, pulmonary artery pressure and right ventricular hypertrophy index were measured and pulmonary artery vessels were studied by light microscope. Then terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)technique was used to detect nucleosomal DNA fragmentation of apoptotic cells.In situ hybridization and RT-PCR were used to detect the expression level of bcl-2 and bax.``Results. The pulmonary artery pressure and right ventricular hypertrophy index of hypoxia group were increased significantly, the pulmonary artery wall of hypoxic group become incrassate than control group. Apoptotic cells can be found in lung with hypoxia or without hypoxia. Compared with control group, apoptotic index of hypoxic group decreased significantly. Through the methods of in situ hybridization and RT-PCR, we found the expression of bcl-2 increased whereas bax decreased significantly in the hypoxic group.``Conclusion. The alternation in bcl-2 and bax expression induced by hypoxia play an important role in the pulmonary artery remodeling which is the main pathologic change of pulmonary hypertension secondary to hypoxia.

  17. THE ROLES OF bcl-2 GENE FAMILY IN THE PULMONARY ARTERY REMODELING OF HYPOXIA PULMONARY HYPERTENSION IN RATS

    Institute of Scientific and Technical Information of China (English)

    王巨; 王凯; 王柏春; 杨成; 王胜发; 梁桃

    2001-01-01

    Objective. To investigate the roles of apeptosis in the pulmonary artery remodeling of pulmonary hypertension secondary to hypoxia and illustrate the relative genes expression. Methods. Thirty rats were divided into hypoxia group(10%O2, 8h/d) and normal control group. On the 15th day of hypoxia, pulmonary artery pressure and fight ventricular hypertrophy index were measured and pulmonary artery vessels were studied by light microscope. Then terminal deoxynucleotidyl transferase-mediateddUTP nick-end labeling(TUNEL)technique was used to detect nucleosomal DNA fragmentation of apeptotic cells.In situ hybridization and RT-PCR were used to detect the expression level of bel-2 and bax. Results. The pulmonary artery pressure and right ventricular hypertrophy index of hypoxia group were increased significantly, the pulmonary artery wall of hypoxic group become incrassate than control group. Apeptotic cells can be found in lung with hypoxia or without hypexia. Compared with control group, apeptotic index of hypoxic group decreased significantly. Through the methods of in situ hybridization and RT-PCR, we found the expression of bel-2 increased whereas bax decreased significantly in the hypoxic group. Conclusion. The alternation in bel-2 and bax expression induced by hypoxia play an important role in the pulmonary artery remodeling which is the main pathologic change of p~monary hypertension secondary to hypoxia.

  18. Diversity and distribution of subterranean bacteria in groundwater at Oklo in Gabon, Africa, as determined by 16S rRNA gene sequencing.

    Science.gov (United States)

    Pedersen, K; Arlinger, J; Hallbeck, L; Pettersson, C

    1996-06-01

    This paper describes how ground water was sampled, DNA extracted, amplified and cloned and how information available in the ribosomal 16S rRNA gene was used for mapping diversity and distribution of subterranean bacteria in groundwater at the Bangombé site in the Oklo region. The results showed that this site was inhabited by a diversified population of bacteria. Each borehole was dominated by species that did not dominate in any of the other boreholes; a result that probably reflects documented differences in the geochemical environment. Two of the sequences obtained were identified at genus level to represent Acinetobacter and Zoogloea, but most of the 44 sequences found were only distantly related to species in the DNA database. The deepest borehole, BAX01 (105 m), had the highest number of bacteria and also total organic carbon (TOC). This borehole harboured only Proteobacteria beta group sequences while sequences related to Proteobacteria beta, gamma and delta groups and Gram-positive bacteria were found in the other four boreholes. Two of the boreholes, BAX02 (34 m) and BAX04 (10 m) had many 16S rRNA gene sequences in common and also had similar counts of bacteria, content of TOC, pH and equal conductivity, suggesting a hydraulic connection between them.

  19. Human neuronal apoptosis secondary to traumatic brain injury and the regulative role of apoptosis-related genes

    Institute of Scientific and Technical Information of China (English)

    杨树源; 雪亮

    2004-01-01

    Objective: To observe human neuronal apoptosis secondary to traumatic brain injury, and to elucidate its regulative mechanism and the change of expression of apoptosis-related genes.Methods: Specimens of brain were collected from cases of traumatic brain injury in humans. The histological and cellular morphology was examined by light and electron microscopy. The extent of DNA injury to cortical neurons was detected by using TUNEL. By in situ hybridisation and immunohistochemistry the mRNA changes and protein expression of Bcl-2, Bax, p53, and caspase 3 p20 subunit were observed.Results: Apoptotic neurons appeared following traumatic brain injury, peaked at 24 hours and lasted for 7 days. In normal brain tissue activated caspase 3 was rare,but a short time after trauma it became activated. The activity peaked at 20-28 hours and remained higher than normal for 5-7 days. There was no expression of Bcl-2 mRNA and Bcl-2 protein in normal brain tissue but 8 hours after injury their expression became evident and then increased, peaked at 2-3 days and remained higher than normal for 5-7 days. The primary expression of Bax-mRNA and Bax protein was high in normal brain tissue. At 20-28 hours they increased and remained high for 2-3 days; on the 7th days they returned to a normal level. In normal brain tissue, p53mRNA and P53 were minimally expressed.Increased expression was detected at the 8th hour, and decreased at 20-28 hours but still remained higher than normal on the 5th day.Conclusions: Following traumatic injury to the human brain, apoptotic neurons appear around the focus of trauma. The mRNA and protein expression of Bcl-2, Bax and p53 and the activity of caspase 3 enzyme are increased.

  20. Immunoglobulin genes

    Energy Technology Data Exchange (ETDEWEB)

    Honjo, T. (Kyoto Univ. (Japan)); Alt, F.W. (Columbia Univ., Dobbs Ferry, NY (USA). Hudson Labs.); Rabbitts, T.H. (Medical Research Council, Cambridge (UK))

    1989-01-01

    This book reports on the structure, function, and expression of the genes encoding antibodies in normal and neoplastic cells. Topics covered are: B Cells; Organization and rearrangement of immunoglobin genes; Immunoglobin genes in disease; Immunoglobin gene expression; and Immunoglobin-related genes.

  1. Evaluation of Bcl-2 Family Gene Expression in Hippocampus of 3, 4-methylenedioxymethamphetamine Treated Rats

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    Hamed Hashemi-Nasl

    2012-01-01

    Full Text Available Objective: 3,4-methylenedioxymethamphetamine (MDMA is an illicit, recreational drugthat causes cellular death and neurotoxicity. This study evaluates the effects of differentdoses of MDMA on the expression of apoptosis–related proteins and genes in the hippocampusof adult rats.Materials and Methods: In this expremental study,a total of 20 male Sprague Dawley rats(200-250 g were treated with MDMA (0, 5, 10, 20 mg/kg i.p. twice daily for 7 days. Sevendays after the last administration of MDMA, the rats were killed. Bax and Bcl-2 genesin addition to protein expressions were detected by western blot and reverse transcriptionpolymerasechain reaction (RT-PCR.Results were analyzed using one-way ANOVA andp≤0.05 was considered statistically significant.Results: Our results showed that MDMA caused dose dependent up-regulation of Baxand down-regulation of Bcl-2 in the hippocampus. There was a significant alteration inbcl-2 and bax genes density.Conclusion: Changes in apoptosis-related proteins and respective genes relating to Baxand Bcl-2 might be involved in the molecular mechanism of MDMA-induced apoptosis.

  2. Changes of NF-κB, Bax and Caspase 3 in Apoptosis lnduced by Ligustrazine Combined with Cis-dichlorodiamine Platinum in Human Gastric Carcinoma SGC-7901 Cell Lines

    Institute of Scientific and Technical Information of China (English)

    Tao HUANG; Liyan Ll; Xiaona GUO; Zhigang GUO; Yalin Zhang

    2015-01-01

    Objective] This study aimed to investigate the mechanism of apoptosis in-duced by ligustrazine (TMP) and cis-dichlorodiamine platinum (DDP) in SGC-7901 cel lines in vitro. [Methods] SGC-7901 cel lines were treated with ligustrazine and DDP alone or combined for 48 h for Western blot analysis, respectively. Western blot analysis was used to determine the expression of proteins involved in apoptosis including NF-κB p65, bax and caspase-3. [Results] The viability of SGC-7901 cel s was inhibited after treated with ligustrazine and/or combined with DDP. The expres-sion of NF-κB P65 protein decreased after treated with drugs, in which the protein decreased significantly in 1.2 mg/ml of TMP combined with 2 μg/ml of DDP group. Meanwhile, we investigated the protein expression of bax and caspase-3. The re-sults showed that the expression of the two proteins increased fol owing with the in-creasing concentration of TMP. [Conclusion] Al the results indicated that ligustrazine combined with DDP could induce the apoptosis of SGC-7901 cel lines, and NF-κB maybe the possible way to induce the cel apoptosis.

  3. Synthetic Bichalcone TSWU-BR23 Induces Apoptosis of Human Colon Cancer HT-29 Cells by p53-Mediated Mitochondrial Oligomerization of BAX/BAK and Lipid Raft Localization of CD95/FADD.

    Science.gov (United States)

    Lin, Meng-Liang; Chen, Shih-Shun; Wu, Tian-Shung

    2015-10-01

    A synthetic bichalcone analog, (E)-1-(3-((4-(4-acetylphenyl)piperazin-1-yl)methyl)-4-hydroxy-5-methoxyphenyl)-3-(pyridin-3-yl)prop-2-en-1-one (TSWU-BR23), has been shown to induce apoptosis in human colon cancer HT-29 cells involving the induction of CD95 and FAS-associated protein death domain (FADD), but its precise mechanism of action has not been fully elucidated. Using cell-surface biotinylation and sucrose density-gradient-based membrane flotation techniques, we showed that the disruption of TSWU-BR23-induced lipid raft localization of CD95/FADD by cholesterol-depleting agent (methyl-β-cyclodextrin) was reversed by cholesterol replenishment. Blockade of p53 expression by short-hairpin RNA (shRNA) suppressed oligomeric Bcl-2-associated x protein (BAX)/Bcl-2 antagonist killer 1 (BAK)-mediated mitochondrial apoptosis but did not inhibit lipid raft localization of CD95/FADD and pro-caspase-8 cleavage induced by TSWU-BR23. Co-expression of p53 shRNA and dominant-negative mutant of FADD completely inhibited TSWU-BR32-induced mitochondrial apoptotic cell death. Collectively, these data demonstrate that TSWU-BR23 leads to HT-29 cell apoptosis by inducing p53-mediated mitochondrial oligomerization of BAX/BAK and the localization of CD95/FADD with lipid rafts at the cell surface.

  4. Microstructure and microwave dielectric properties of Ba6-3xSm8+2xTi18Os4ceramics with various BaxSr1-xTiO3 additions

    Institute of Scientific and Technical Information of China (English)

    ZHOU Lingling; ZHOU Hongqing; SHAO Hui; ZHU Haikui

    2012-01-01

    The Ba6-3xSm8+2xTi18O54 (x=2/3) microwave dielectric ceramics were prepared by traditional solid sate reaction technique.The experiment was based on the Ba6-3xSm8+2xTi18O54 (BST) microwave dielectric ceramics doped with a certain amount of Bi2O3,then the effects of BaxSr1-xTiO3 additives on the structure and microwave dielectric properties of Ba6-3xSm8+2xTi18O54 ceramics were investigated using X-ray diffraction and scanning electron microscopy.In this study,the small amount substitution of Sr for Ba was effective for the microwave dielectric properties of BST,especially the τf could be tuned to near zero.The result showed that the BST possessed excellent dielectric properties when the addition of Bi2O3 and BaxSr1-xTiO3 was 1 wt.% respectively:εr=79.6,Qf=10789 GHz,τf=-1.5 ppm/℃.

  5. Synergistic Effect of Subtoxic-dose Cisplatin and TRAIL to Mediate Apoptosis by Down-regulating Decoy Receptor 2 and Up-regulating Caspase-8, Caspase-9 and Bax Expression on NCI-H460 and A549 Cells

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    Xiaoyan Zhang

    2013-05-01

    Full Text Available Objective(s: Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL can selectively induce apoptosis in tumor cells, more than half of tumors including non-small cell lung cancer (NSCLC exhibit TRAIL-resistance. The purpose of this study was to determine whether subtoxic-dose cisplatin and TRAIL could synergistically enhance apoptosis on NSCLC cells and investigate its underlying mechanisms. Materials and Methods:NCI-H460 and A549 cells were treated with TRAIL alone, cisplatin alone or combination treatment in this study. The cytotoxicity was evaluated according to Sulforhodamine B assay, and apoptosis was examined using Hoechst 33342 staining and flow cytometry. The mRNA and protein levels of TRAIL receptors and apoptotic proteins including caspase-8, caspase-9, Bcl-2 and Bax were determined by RT-PCR and Western blotting, respectively. Results:Our results showed that NCI-H460 cells were sensitive to TRAIL, whereas A549 cells were resistant. However, subtoxic-dose cisplatin could enhance the both cells to TRAIL-mediated cell proliferation inhibition and apoptosis. The underlying mechanisms might be associated with the down-regulation of DcR2 and up-regulation of Caspase-8, Caspase-9 and Bax. Conclusion:Subtoxic-dose cisplatin could enhance both TRAIL- sensitive and TRAIL- resistant NSCLC cells to TRAIL-mediated apoptosis. These findings motivated further studies to evaluate such a combinatory therapeutic strategy against NSCLC in the animal models.

  6. Dioscorealide B from the traditional Thai medicine Hua-Khao-Yen induces apoptosis in MCF-7 human breast cancer cells via modulation of Bax, Bak and Bcl-2 protein expression.

    Science.gov (United States)

    Saekoo, Jiraporn; Graidist, Potchanapond; Leeanansaksiri, Wilairat; Dechsukum, Chavaboon; Itharat, Arunporn

    2010-12-01

    Dioscorealide B is a pharmacologically active compound from the rhizome of the Thai medicinal plant Dioscorea membranacea. Here, we demonstrated that in vitro treatment of dioscorealide B resulted in a cytotoxic effect on MCF-7 human breast cancer cells (IC50 = 2.82 microM). To determine whether this compound induces apoptosis in MCF-7, the Annexin V assay was performed. The data showed that the number of apoptotic cells were increased 7-12 folds over that of the control cells after treatment with various concentrations of dioscorealide B (3, 6 and 12 microM) for 24 hours. Dioscorealide B-induced apoptosis was associated with modulation of the multidomain Bcl-2 family members Bax, Bak and Bcl-2. After treatment with 3 microM dioscorealide B, acceleration of the level of proapoptotic proteins Bax and Bak were observed at 6 hours and 12 hours, respectively, while the decrease in the expression of antiapoptotic protein Bcl-2 was observed 3 hours after the treatment. These effects of dioscorealide B might result in the activation of caspase-8, -9 and -7, which lead to apoptosis in MCF-7 cells. Taken together, the results of this study provide evidence that dioscorealide B possesses an antitumor property against human breast cancer cells and thus provide the molecular basis for the further development of dioscorealide B as a novel chemotherapeutic agent for breast cancer treatment. PMID:21299121

  7. Protective Effects of Scutellarin on Type II Diabetes Mellitus-Induced Testicular Damages Related to Reactive Oxygen Species/Bcl-2/Bax and Reactive Oxygen Species/Microcirculation/Staving Pathway in Diabetic Rat

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    Lingli Long

    2015-01-01

    Full Text Available The goal of our study is to evaluate the effect of Scutellarin on type II diabetes-induced testicular disorder and show the mechanism of Scutellarin’s action. We used streptozotocin and high-fat diet to establish type II diabetic rat model. TUNEL and haematoxylin and eosin staining were used to evaluate the testicular apoptotic cells and morphologic changes. Immunohistochemical staining was used to measure the expression level of vascular endothelial growth factor and blood vessel density in testes. Oxidative stress in testes and epididymis was tested by fluorescence spectrophotometer and ELISA. The expression of Bcl-2/Bax and blood flow rate in testicular vessels were measured by western blot and Doppler. Our results for the first time showed that hyperglycemia induced apoptotic cells and morphologic impairments in testes of rats, while administration of Scutellarin can significantly inhibit these damages. This effect of Scutellarin is controlled by two apoptotic triggers: ROS/Bcl-2/Bax and ROS/microcirculation/starving pathway.

  8. 新的细胞凋亡抑制基因survivin在垂体腺瘤中的表达及其与caspase-3表达相关性的研究%Apoptosis- regulating protein,survivin:expression and relationship with caspase- 3 protein in pituitary adenomas

    Institute of Scientific and Technical Information of China (English)

    马杰; 石智勇; 魏冰; 乔思杰

    2002-01-01

    Objective To study the expression of a inhibitor gene,survivin,in pituitary adenomas and its relationship with the expression of caspase- 3 gene.Method Using streptavidin- biotin peroxidase(SP),the expression of surviving,caspase- 3 protein was examined in 8 normal pituitary tissue and 38 pituitary adenomas.Result Survivin was expressed in 23 of 38 cases of pituitary adenomas,in contrast,normal pituitary tissue did not express surviving.Positive expression rate of PRL group,GH group,Mixed group in pituitary adenomas was 12/17,7/13,4/8,respectively.Overexpression of surviving was not related with the tumor type(P >0.05).Meanwhile,surviving positive rate in caspase- 3 positive and negative groups was 5/14,79% .The result showed it was strongly associated with caspase- 3 expression(P<0.05).Conclusions Apoptosis suppression by surviving play an important role in apoptosis of pituitary adenomas.Survivin could be a new diagnostic/therapeutic target in pituitary adenomas.

  9. 化癥散积颗粒对小鼠原位肝癌Bcl-2/Bax表达的影响%The effects of Huazhengsanjikeli on Bcl-2/Bax expression in mouse of primary Hepatic carcinoma

    Institute of Scientific and Technical Information of China (English)

    金学洙; 李超英; 周磊; 邢美茜; 刘铁军

    2012-01-01

    目的 通过建立小鼠原位肝癌模型,给予化癥散积颗粒灌胃,探讨其对小鼠原位肝癌Bcl-2/Bax表达的影响.方法 将H22瘤株直接注射到肝脏的方法建立小鼠原位肝癌模型;分组:模型组、阳性对照组、化癥散积颗粒高、中、低剂量组;采用TUNEL染色法检测凋亡细胞;RT-PCR和Western-Blot分别检测肿瘤组织Bcl-2和Bax mRNA和蛋白表达差异.结果 阳性对照组(又称斑蝥组)和中、高剂量组中药可以诱导肝癌细胞发生凋亡;可以显著增加肿瘤组织中Bax(P<0.05),同时Bcl-2表达与对照组相比明显降低(P<0.05),低剂量组Bcl-2和Bax表达与对照组相比无明显差异(P>0.05).结论 斑蝥组和中、高剂量组中药诱导肝癌细胞发生凋亡可能是通过抑制Bcl-2基因表达,同时增加Bax表达,从而引发一系列凋亡级联反应.%Objective To study the antitumor mechanism of Granules scattered plot of disease through the insitu-liver cancer model,in order to offer some evidence for clinical Chinese crude drug treatment. Methods Use H22 cells inject liver to build liver caner modcr; Detect apoptosis cells in tumor by TUNKL; Detect the expression difference of Bel-2/Bax by RT-PCR and Western-blot;Results The number of apoptosis cells were significantly increased in Cantharidin,medium and high dose Granules scattered plot of disease groups than in control and low dose groups; The mRNA and protein expression of Bax can be significantly up-rcgulation(P0. 05). Conclusion Cantharidin,medium and high dose Granules scattered plot of disease can induce H22 liver cancr cells to apoptosis; Granules? Scattered plot of disease can be induce liver cancer cells to apoptosis through inhibited the expression of Bcl-2 and up-regulated the expression of Bax at the same time.

  10. 奥曲肽对人肝星状细胞凋亡及Bcl-2/Bax表达的影响%Effects of octreotide on the apoptosis of human HSCs and expression of Bcl-2/Bax in HSCs

    Institute of Scientific and Technical Information of China (English)

    李春艳; 贾丽萍; 石蕾; 周贤

    2015-01-01

    Objective To investigate the effects of octreotide on the apoptosis of human hepatic stellate cells (HSCs) and expression of Bcl-2/Bax in HSCs,and to reveal the mechanism underlying octreotide against hepatic fibrosis. Methods HSCs lines (HSC-LX2) were incubated with different concentrations of octreotide for 24 and 48 hours. Cell apoptosis was evaluated by Fitc-tunel fluorescence staining. Bcl-2 and Bax protein exoression in HSC-LX2 was detected by immunocytochemistry. Meanwhile, Bcl-2 protein of HSC-LX2 were detected by Western blot assay. Results Octreotide could promote the apoptosis of HSC-LX2, and the apoptosis rate was significantly increased with the concentration of octreotide(P < 0.05). The HSC-LX2 were incubated with the same concentration of octreotide for 24 and 48 hours, the cell apoptosis rate of 48-hour octreotide treatment was significantly higher than that of 24-hour octreotide treatment (P < 0.05). The immunocytochemistry result indicated that octreotide could significantly decrease Bcl-2 expression and increase Bax expression in HSC-LX2 (P<0.05); Western blot assay showed that octreotide could also significantly inhibit Bcl-2 expression in HSC-LX2 (P<0.05). Conclusions Octreotide could induce the apoptosis of HSCs in a dose-and time-dependent manner, the mechanism of octreotide inducing HSCs apoptosis might be associated with down-regulation of Bcl-2 and upregulation of Bax in HSC.%目的:观察奥曲肽对活化人肝星状细胞凋亡和凋亡相关蛋白Bcl-2/Bax 表达的影响,探讨奥曲肽抗肝纤维化可能的作用机制。方法:不同浓度的奥曲肽作用于传代的人肝星状细胞株(HSC-LX2)24 h、48 h后,应用FITC-TUNEL检测各组细胞凋亡,应用免疫细胞化学法检测HSC-LX2中Bcl-2、Bax蛋白表达,应用Western-blot法检测HSC-LX2中Bcl-2蛋白表达。结果:奥曲肽可促进HSC-LX2细胞调亡,细胞凋亡率随奥曲肽浓度增加而增高(P <0.05);与24 h比较,相

  11. Bcl-2 and Bax expression in mice with collagen-induced arthritis following heterogenic umbilical cord blood stem cell transplantation%异种脐血干细胞移植胶原性关节炎小鼠Bcl-2和Bax 的表达

    Institute of Scientific and Technical Information of China (English)

    牛广华; 高玉洁; 孙旭; 都静; 明彩荣; 高明利

    2012-01-01

    BACKGROUND: The occurrenceand development of rheumatoid arthritis were strongly associated with unbalance proliferation and apoptosis of synovial cells and lymphocytes. Some synovial cell apoptosis was abnormal. OBJECTIVE: To observe effects of heterogenic double umbilical cord blood stem cell transplantation on Bcl-2 and Bax expression in mice with type Ⅱcollagen-induced arthritis. METHODS: C57BL/6(H-2b) mice were induced with Freund's complete adjuvant and type Ⅱcollagen to establish mouse models of type Ⅱcollagen-induced arthritis.At 2days after secondary immunity incubation, mice were injected with saline via tail vein in the model and normal control groups. Umbilical cord blood hemopoietic stem cells were injected into mouse tail vein in the UBSCs transplantation groups (single dose:2×106/50 g; double dose: 1×106/50 g each, totally 2×106/50 g). Mice were intragastrically administrated methotrexate in the methotrexate positive control group, 0.017 5 g/kg once, once every 5 days, totally six times. RESULTS AND CONCLUSION: Joint tissue below knee and elbow was obtained at42 days following transplantation. Histopathology displayed that smooth and glossy articular surface, no inflammatory cell infiltration in the synovial layer and normal chondrocytes in the normal control group. Hyperplasia, a lot of inflammatory cell infiltration and damaged cartilage surface were visible in the model group. Slight hyperplasia and a little inflammatory cell infiltration were detectable in the methotrexate positive controlgroup and single UBSCs transplantation group. Double UBSCs transplantation group exhibited smooth and glossy articular cartilage surface, no damage, a little inflammatory cell infiltration. Immunohistochemistry demonstrated that Bcl-2 and Bax expression was lower in the double UBSCs transplantation group compared with single UBSCs transplantation group (P 0.05). Results suggest that double umbilical cord blood stem cells can induce synovial cell

  12. Disruption of the ECM33 Gene in Candida albicans Prevents Biofilm Formation, Engineered Human Oral Mucosa Tissue Damage and Gingival Cell Necrosis/Apoptosis

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    Mahmoud Rouabhia

    2012-01-01

    Full Text Available In this study we demonstrated that ΔCaecm33 double mutant showed reduced biofilm formation and causes less damage to gingival mucosa tissues. This was confirmed by the reduced level of necrotic cells and Bax/Bcl2 gene expression as apoptotic markers. In contrast, parental and Caecm33 mutant strains decreased basement membrane protein production (laminin 5 and type IV collagen. We thus propose that ECM33 gene/protein represents a novel target for the prevention and treatment of infections caused by Candida.

  13. Caractérisation structurale de l'adsorption des isomères para- et meta- du xylène dans la zéolithe de type faujasite BaX Structural Characterization of Para- and Meta- Xylene in Bax Zeolite

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    Mellot C.

    2006-11-01

    Full Text Available La séparation du para-xylène des isomères aromatiques en C8 est réalisée industriellement grâce à l'adsorption sur tamis moléculaire zéolithique. Une amélioration des propriétés de séparation des tamis, et en particulier de leur sélectivité, nécessite, entre autres, une bonne connaissance des interactions entre les molécules d'adsorbat et la structure zéolithique. Pour ce faire, nous avons fait appel à deux techniques de caractérisation physico-chimique : la diffraction des neutrons et la spectroscopie infrarouge. Nous avons étudié une zéolithe BaX de type faujasite sur laquelle nous avons adsorbé, à l'état de corps purs, les isomères para- et méta- du xylène. Cette zéolithe est connue industriellement pour ses propriétés sélectives performantes pour le paraxylène. Les analyses ont été réalisées en considérant tout d'abord un faible taux de remplissage, voisin d'une molécule par supercage de zéolithe, et ensuite à saturation où la zéolithe contient sensiblement trois molécules par supercage. La diffraction des neutrons permet, à basse température, de localiser les molécules dans la zéolithe et de préciser leur interaction avec le cation Ba²+. La spectroscopie infrarouge permet une étude des caractéristiques vibrationnelles de l'adsorbat en fonction du taux de recouvrement. Une synthèse des résultats obtenus à l'aide de ces deux méthodes d'investigation nous a permis de dégager un modèle de remplissage des supercages pour les deux isomères considérés. Ainsi, des différences significatives sont mises en évidence. En ce qui concerne le para-xylène, pour un taux de remplissage inférieur à deux molécules par supercage, les molécules de para-xylène viennent se positionner au voisinage des cations en site SII de la supercage. La troisième molécule de para-xylène introduite dans la zéolithe n'est pas en interaction avec un cation Ba²+ et sera localisée dans un nouveau site F

  14. Differential Gene Expression Profiling of Dystrophic Dog Muscle after MuStem Cell Transplantation.

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    Florence Robriquet

    Full Text Available Several adult stem cell populations exhibit myogenic regenerative potential, thus representing attractive candidates for therapeutic approaches of neuromuscular diseases such as Duchenne Muscular Dystrophy (DMD. We have recently shown that systemic delivery of MuStem cells, skeletal muscle-resident stem cells isolated in healthy dog, generates the remodelling of muscle tissue and gives rise to striking clinical benefits in Golden Retriever Muscular Dystrophy (GRMD dog. This global effect, which is observed in the clinically relevant DMD animal model, leads us to question here the molecular pathways that are impacted by MuStem cell transplantation. To address this issue, we compare the global gene expression profile between healthy, GRMD and MuStem cell treated GRMD dog muscle, four months after allogenic MuStem cell transplantation.In the dystrophic context of the GRMD dog, disease-related deregulation is observed in the case of 282 genes related to various processes such as inflammatory response, regeneration, calcium ion binding, extracellular matrix organization, metabolism and apoptosis regulation. Importantly, we reveal the impact of MuStem cell transplantation on several molecular and cellular pathways based on a selection of 31 genes displaying signals specifically modulated by the treatment. Concomitant with a diffuse dystrophin expression, a histological remodelling and a stabilization of GRMD dog clinical status, we show that cell delivery is associated with an up-regulation of genes reflecting a sustained enhancement of muscle regeneration. We also identify a decreased mRNA expression of a set of genes having metabolic functions associated with lipid homeostasis and energy. Interestingly, ubiquitin-mediated protein degradation is highly enhanced in GRMD dog muscle after systemic delivery of MuStem cells.Overall, our results provide the first high-throughput characterization of GRMD dog muscle and throw new light on the complex

  15. Altered gene expression profiles of NIH3T3 cells regulated by human lung cancer associated gene CT120

    Institute of Scientific and Technical Information of China (English)

    Xiang Huo HE; Jin Jun LI; Yi Hu XIE; Yun Tian TANG; Gen Fu YAO; Wen Xin QIN; Da Fang WAN; Jian Ren GU

    2004-01-01

    CT120, a novel membrane-associated gene implicated in lung carcinogenesis, was previously identified from chromosome 17p13.3 locus, a hot mutation spot involved in human malignancies. In the present study, we further determined that CT120 ectopic expression could promote cell proliferation activity of NIH3T3 cells using MTS assay, and monitored the downstream effects of CT120 in NIH3T3 cells with Atlas mouse cDNA expression arrays. Among 588known genes, 133 genes were found to be upregulated or downregulated by CT120. Two major signaling pathways involved in cell proliferation, cell survival and anti-apoptosis were overexpressed and activated in response to CT120:One is the Raf/MEK/Erk signal cascades and the other is the PI3K/Akt signal cascades, suggesting that CT120 might contribute, at least in part, to the constitutively activation of Erk and Akt in human lung caner cells. In addition, some tumor metastasis associated genes cathepsin B, cathepsin D, cathepsin L, MMP-2/TIMP-2 were also upregulated by CT120, upon which CT120 might be involved in tumor invasiveness and metastasis. In addition, CT120 might play an important role in tumor progression through modulating the expression of some candidate "Lung Tumor Progression"genes including B-Raf, Rab-2, BAX, BAG-1, YB-1, and Cdc42.

  16. Gene expression patterns in heterozygous Plk4 murine embryonic fibroblasts

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    Nantais Jordan

    2009-07-01

    Full Text Available Abstract Background The polo-like kinases (Plks are a group of serine/threonine kinases which have roles in many aspects of cellular function including the regulation of mitotic activity and cellular stress responses. This study focuses on Plk4, the most divergent member of the Plk family, which is necessary for proper cellular proliferation. More specifically, alterations in Plk4 levels cause significantly adverse mitotic defects including abnormal centrosome duplication and aberrant mitotic spindle formation. We sought to clarify the effect of reduced Plk4 levels on the cell by examining transcript profiles of Plk4 wild-type and heterozygous mouse embryonic fibroblasts (MEFs. Subsequently, the levels of several key proteins involved in the DNA damage response were examined. Results 143 genes were found to be significantly up-regulated in the heterozygous MEFs compared to their wild-type counterparts, while conversely, 9 genes were down-regulated. Numerous genes with increased transcript levels in heterozygous MEFs were identified to be involved in p53-dependent pathways. Furthermore, examination of the promoter regions of all up- and down-regulated genes revealed that the majority contained putative p53 responsive elements. An analysis of transcript levels in MEFs after exposure to either ionizing or ultraviolet radiation revealed a significant change between wild type and heterozygous MEFS for Plk4 transcript levels upon only UV exposure. Furthermore, changes in protein levels of several important cell check-point and apoptosis regulators were examined, including p53, Chk1, Chk2, Cdc25C and p21. In heterozygous MEFs, p53, p21 and Chk2 protein levels were at significantly higher levels. Furthermore, p53 activity was increased 5 fold in the Plk4 heterozygous MEFs. Conclusion Global transcript profiles and levels of key proteins involved in cellular proliferation and DNA damage pathways were examined in wild-type and Plk4 heterozygous MEFs. It

  17. 溴隐亭对大鼠催乳素瘤细胞表达bcl-2、Bax的影响%Effect of bromocriptine on expression of Bax and bcl-2 in rat prolactinoma

    Institute of Scientific and Technical Information of China (English)

    杨雪梅; 徐春; 梁立武; 程海梅

    2012-01-01

    目的 研究溴隐亭对大鼠催乳素(prolactinoma,PRL)瘤表达bcl-2、Bax的影响.方法 (1)用皮下植入17β-雌二醇的方法制备大鼠催乳素瘤模型,同时设立10只大鼠为对照组;(2)将成功诱发出催乳素瘤的大鼠随机分2组,分别给予自来水(安慰剂组)、溴隐亭(溴隐亭组)灌胃,对照组也给予安慰剂灌胃,用药4周后处死动物,垂体称重,测定PRL、Bax、bcl-2的表达水平.结果 (1)17β-雌二醇组大鼠血清PRL水平[(4236.9±416.9) vs (121.2±12.8) ng/ml]和垂体重量[(62.0±5.1) vs (13.8±1.2) mg]均明显高于对照组(P<0.01),证实17β-雌二醇组成功诱发出大鼠泌乳素瘤.(2)溴隐亭组血清PRL水平和垂体重量低于安慰剂组(P<0.01),bcl-2表达水平较安慰剂组明显下降[(1.8±0.5) vs (4.0±0.6),P<0.01],Bax表达水平明显增高[(4.5±0.6) vs ( 1.0±0.3),P<0.01].结论 抑制bcl-2的表达,刺激Bax的表达,从而促进催乳素瘤细胞的凋亡可能是溴隐亭抗催乳素瘤的重要机制之一.%Objective To study the effect of bromocriptine on expression of Bax and bcl -2 in rat prolactinoma. Methods Firstly, to develop prolactinoma rats model. Adult Wistar rats were divided into two groups at random. The rats in control group were subscutaneously implanted with a blank implant . Rats in 17 p - estradiol group were implanted with 17 p - estradiol - containing implants. Secondly, Rats in 17p - estradiol group were divided into two groups at random, i. e. placebo group and bromocriptine group. Water was given to rats in placebo group. Bromocriptine was orally adminstered to rats in bromocriptine group. After 4 weeks of treatment, all the animals were sacrificed. Each pituitary gland was weighed. Serum prolactin(PRL) levels were measured by RIA. Expression level of Bax and bcl -2 in pituitary tissue were measured by Western blotting. Results (1) The weights of pituitary gland and PRL levels in 17p - estradiol group were significantly higher than those

  18. The Influence of Matrine on Apoptosis and Expression of Bax and Bci-2 in Colorectal Cancer Cells%苦参碱对大肠癌细胞凋亡及Bax、Bcl-2表达的影响

    Institute of Scientific and Technical Information of China (English)

    王雷; 刘明

    2012-01-01

    [Purpose] To investigate the effect of Matrine on proliferation inhibition, apoptotic and Bax and Bcl-2 expression in human colorectal cancer cell line Lovo. [Methods] Lovo cells cultured in vitro were interfered with 0.05-1.6mg/ml different concentration of Matrine. The proliferation inhibition effect on Lovo cells was observed by MTT method. Apoptosis induction effect on Lovo cells was detected by DNA ladder, flow cytometer and TUNEL staining. The expression of Bcl-2 and Bax proteins correlated with apoptosis were detected by Western Blot assay. [Results] After being exposed to Matrine (0.05-1.6mg/ml) for 24 and 48h, the proliferation of Lovo cells was inhibited in a dose-time dependent manner. DNA ladder, Annexin V-PI method and TUNEL staining showed Matrine was obviously increased along with Matrine concentration increased. The expression of pro-apoptotic protein Bax was increased, while anti-apoptotic protein Bcl-2 was decreased as Matrine doses increased. [Conclusion] Matrine can inhibit proliferation and induction of apoptosis in colorectal cancer cells. Increased expression of Bax and decreased expression of Bcl-2 might involve in Matrine-induced apoptosis.%[目的]探讨苦参碱对人大肠癌Lovo细胞增殖抑制和凋亡诱导作用及其对Bax、Bcl-2表达的影响.[方法] 0.05~l.6mg/ml不同浓度苦参碱作用Lovo细胞,采用MTT法检测苦参碱对大肠癌Lovo细胞增殖抑制作用,DNA ladder、AnnexinV -PI法及TUNEL染色检测细胞凋亡,Western Blot法检测凋亡相关蛋白Bax、Bcl-2表达的变化.[结果]0.05~1.6mg/ml苦参碱处理Lovo细胞24h或48h后,细胞增殖均明显受抑制;DNA ladder、Annexin V-PI法及TUNEL染色检测结果显示苦参碱呈时间、剂量依赖性诱导细胞凋亡;促凋亡蛋白Bax随着苦参碱剂量增加表达增加,抗凋亡蛋白Bcl-2随着苦参碱剂量增加表达减少.[结论]苦参碱具有抑制大肠癌细胞增殖,诱导其凋亡的作用.苦参碱诱导大肠癌细胞凋

  19. Relationship between ferroelectric properties and local structure of Pb1-xBaxZr0.40Ti0.60O3 ceramic materials studied by X-ray absorption and Raman spectroscopies

    Science.gov (United States)

    Mesquita, Alexandre; Michalowicz, Alain; Moscovici, Jacques; Pizani, Paulo Sergio; Mastelaro, Valmor Roberto

    2016-08-01

    This paper reports on the structural characterization of Pb1-xBaxZr0.40Ti0.60O3 (PBZT) ferroelectric ceramic compositions prepared by the conventional solid state reaction method. X-ray absorption spectroscopy (XAS) and Raman spectroscopy were used in the probing of the local structure of PBZT samples that exhibit a normal or relaxor ferroelectric behavior. They showed a considerable local disorder around Zr and Pb atoms in the samples of tetragonal or cubic long-range order symmetry. The intensity of the E(TO3) mode in the Raman spectra of PBZT relaxor samples remains constant at temperatures lower than Tm, which has proven the stabilization of the correlation process between nanodomains.

  20. Drinking-Water Arsenic Exposure Modulates Gene Expression in Human Lymphocytes from a U.S. Population

    Science.gov (United States)

    Andrew, Angeline S.; Jewell, David A.; Mason, Rebecca A.; Whitfield, Michael L.; Moore, Jason H.; Karagas, Margaret R.

    2008-01-01

    Background Arsenic exposure impairs development and can lead to cancer, cardiovascular disease, and diabetes. The mechanism underlying these effects remains unknown. Primarily because of geologic sources of contamination, drinking-water arsenic levels are above the current recommended maximum contaminant level of 10 μg/L in the northeastern, western, and north central regions of the United States. Objectives We investigated the effects of arsenic exposure, defined by internal biomarkers at levels relevant to the United States and similarly exposed populations, on gene expression. Methods We conducted separate Affymetrix microarray-based genomewide analyses of expression patterns. Peripheral blood lymphocyte samples from 21 controls interviewed (1999–2002) as part of a case–control study in New Hampshire were selected based on high- versus low-level arsenic exposure levels. Results The biologic functions of the transcripts that showed statistically significant abundance differences between high- and low-arsenic exposure groups included an overrepresentation of genes involved in defense response, immune function, cell growth, apoptosis, regulation of cell cycle, T-cell receptor signaling pathway, and diabetes. Notably, the high-arsenic exposure group exhibited higher levels of several killer cell immunoglobulin-like receptors that inhibit natural killer cell activity. Conclusions These findings define biologic changes that occur with chronic arsenic exposure in humans and provide leads and potential targets for understanding and monitoring the pathogenesis of arsenic-induced diseases. PMID:18414638

  1. Methoxychlor and triclosan stimulates ovarian cancer growth by regulating cell cycle- and apoptosis-related genes via an estrogen receptor-dependent pathway.

    Science.gov (United States)

    Kim, Joo-Young; Yi, Bo-Rim; Go, Ryeo-Eun; Hwang, Kyung-A; Nam, Ki-Hoan; Choi, Kyung-Chul

    2014-05-01

    Methoxychlor and triclosan are emergent or suspected endocrine-disrupting chemicals (EDCs). Methoxychlor [MXC; 1,1,1-trichlor-2,2-bis (4-methoxyphenyl) ethane] is an organochlorine pesticide that has been primarily used since dichlorodiphenyltrichloroethane (DDT) was banned. In addition, triclosan (TCS) is used as a common component of soaps, deodorants, toothpastes, and other hygiene products at concentrations up to 0.3%. In the present study, the potential impact of MXC and TCS on ovarian cancer cell growth and underlying mechanism(s) was examined following their treatments in BG-1 ovarian cancer cells. As results, MXC and TCS induced BG-1 cell growth via regulating cyclin D1, p21 and Bax genes related with cell cycle and apoptosis. A methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay confirmed that the proliferation of BG-1 ovarian cancer cells was stimulated by MXC (10(-6), 10(-7), 10(-8), and 10(-9)M) or TCS (10(-6), 10(-7), 10(-8), and 10(-9)M). Treatment of BG-1 cells with MXC or TCS resulted in the upregulation of cyclin D1 and downregulation of p21 and Bax transcriptions. In addition, the protein level of cyclin D1 was increased by MXC or TCS while p21 and Bax protein levels appeared to be reduced in these cells. Furthermore, MXC- or TCS-induced alterations of these genes were reversed in the presence of ICI 182,780 (10(-7)M), suggesting that the changes in these gene expressions may be regulated by an ER-dependent signaling pathway. In conclusion, the results of our investigation indicate that two potential EDCs, MXC and TCS, may stimulate ovarian cancer growth by regulating cell cycle- and apoptosis-related genes via an ER-dependent pathway.

  2. Influence of oxidative stress on apoptosis and expression of bax and bcl-2 of enterocytes in burn rats with delayed resuscitation on the plateau%高原地区烧伤后延迟复苏氧化应激对大鼠肠上皮细胞凋亡及bax和bcl-2基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    周文军; 张诚; 刘毅; 刘萍; 马明; 张世范

    2009-01-01

    Objective To explore influence of oxidative stress reaction on apoptosis rate and expres-sion of apoptosis-related genes bax and bcl-2 of enterocytes in severely burned rats with delayed resuscitation on the plateau. Methods One hundred and twenty rats subjected to 30% TBSA full-thickness scald on the back were derided into plateau experimental group (PE, altitude 3840 m) and Lanzhou experimental group (LE, altitude 1517 m). Then LE and PE groups were subdivided into Lanzhou immediate fluid resus-citation group (LIFR, with immediate intraperitoneal injection of isotonic saline after scald, 40 mL/kg), Lanzhou delayed fluid resuscitation group [LDFR, with intraperitoneal injection of isotonic saline at 6 post scald hour (PSH), 40 mL/kg], and plateau immediate fluid resuscitation group (PIFR, with immediate in-traperitoneal injection of isotonic saline after scald, 40 mL/kg), plateau delayed fluid resuscitation group (PDFR, with intraperitoneal injection of isotonic saline at 6 PSH, 40 mL/kg). Another 12 rats were divided into Lanzhou sham scald group (LS) and plateau sham scald group (PS), with 6 rats in each group. Rats in LS and PS groups were sham scalded in a water bath for 15 s without fluid infusion. Rats were sacrificed at 6, 12, 24, 48, 72 PSH for collection of small intestine samples to determine the contents of malonaldehyde (MDA) and total hydrosulfide (TSH). The apoptosis of enterocytes was determined by TUNEL, and the ex-pression of bax and bcl-2 in epithelial cells were observed by immunohistochemical method. Intestinal sample of LS and PS groups were collected to determine the contents of MDA and TSH. Results After being scal-ded, content of MDA in intestinal tissue of rats in LDFR group and PDFR group was respectively greater than that in LIFR group and PIFR group (P<0.05 or P<0.01). Intestinal tissue content of MDA of rats in LDFR group (9.8±4.0 nmol/mg) and PDFR group (10.2±1.3 nmol/mg) was respectively greater than that in LIFR group (9.5±2

  3. Gene expression

    International Nuclear Information System (INIS)

    We prepared probes for isolating functional pieces of the metallothionein locus. The probes enabled a variety of experiments, eventually revealing two mechanisms for metallothionein gene expression, the order of the DNA coding units at the locus, and the location of the gene site in its chromosome. Once the switch regulating metallothionein synthesis was located, it could be joined by recombinant DNA methods to other, unrelated genes, then reintroduced into cells by gene-transfer techniques. The expression of these recombinant genes could then be induced by exposing the cells to Zn2+ or Cd2+. We would thus take advantage of the clearly defined switching properties of the metallothionein gene to manipulate the expression of other, perhaps normally constitutive, genes. Already, despite an incomplete understanding of how the regulatory switch of the metallothionein locus operates, such experiments have been performed successfully

  4. Expression of p53 Target Genes in the Early Phase of Long-Term Potentiation in the Rat Hippocampal CA1 Area

    Science.gov (United States)

    Pustylnyak, Vladimir O.; Lisachev, Pavel D.; Shtark, Mark B.

    2015-01-01

    Gene expression plays an important role in the mechanisms of long-term potentiation (LTP), which is a widely accepted experimental model of synaptic plasticity. We have studied the expression of at least 50 genes that are transcriptionally regulated by p53, as well as other genes that are related to p53-dependent processes, in the early phase of LTP. Within 30 min after Schaffer collaterals (SC) tetanization, increases in the mRNA and protein levels of Bax, which are upregulated by p53, and a decrease in the mRNA and protein levels of Bcl2, which are downregulated by p53, were observed. The inhibition of Mdm2 by nutlin-3 increased the basal p53 protein level and rescued its tetanization-induced depletion, which suggested the involvement of Mdm2 in the control over p53 during LTP. Furthermore, nutlin-3 caused an increase in the basal expression of Bax and a decrease in the basal expression of Bcl2, whereas tetanization-induced changes in their expression were occluded. These results support the hypothesis that p53 may be involved in transcriptional regulation during the early phase of LTP. We hope that the presented data may aid in the understanding of the contribution of p53 and related genes in the processes that are associated with synaptic plasticity. PMID:25767724

  5. Expression of p53 Target Genes in the Early Phase of Long-Term Potentiation in the Rat Hippocampal CA1 Area

    Directory of Open Access Journals (Sweden)

    Vladimir O. Pustylnyak

    2015-01-01

    Full Text Available Gene expression plays an important role in the mechanisms of long-term potentiation (LTP, which is a widely accepted experimental model of synaptic plasticity. We have studied the expression of at least 50 genes that are transcriptionally regulated by p53, as well as other genes that are related to p53-dependent processes, in the early phase of LTP. Within 30 min after Schaffer collaterals (SC tetanization, increases in the mRNA and protein levels of Bax, which are upregulated by p53, and a decrease in the mRNA and protein levels of Bcl2, which are downregulated by p53, were observed. The inhibition of Mdm2 by nutlin-3 increased the basal p53 protein level and rescued its tetanization-induced depletion, which suggested the involvement of Mdm2 in the control over p53 during LTP. Furthermore, nutlin-3 caused an increase in the basal expression of Bax and a decrease in the basal expression of Bcl2, whereas tetanization-induced changes in their expression were occluded. These results support the hypothesis that p53 may be involved in transcriptional regulation during the early phase of LTP. We hope that the presented data may aid in the understanding of the contribution of p53 and related genes in the processes that are associated with synaptic plasticity.

  6. Effect of acrylonitrile on the expression of Bcl-2 and Bax in spermatogenic cell of mice%丙烯腈对小鼠生精细胞Bcl-2、Bax蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    裴凌云; 马国燕; 金娜; 陈亚; 冯玉娟; 党瑜慧; 薛红丽; 李芝兰

    2012-01-01

    Objective To explore the effect of acrylonitrile exposure on the expression of Bcl-2 and Bax in mice spermatogenic cells. Methods Based on body weight, 250 SPF Kunming adult male mice were randomly divided into five groups; negative control group (normal saline 0. 01ml/g), three AN exposure groups (intraperitoneal injection of 1.25, 2. 50 or 5. 00 mg/kg of AN daily for 5 days, respectively) and positive control group (intraperitoneal injection of cyclophosphamide 40mg/kg) . Mice were killed in the 7th, 14th, 21st, 28th and 35th day after the first exposure by cervical dislocation. Immunohistochemical method ( SABC ) was used to detect the expression of Bcl-2 and Bax protein in spermatogenic cells. Results The average optical density values of Bcl-2 at five time points of the AN 2. 50 mg/kg group and the 21th day point of the AN 1. 25 mg/kg group were significantly lower than the negative control group ( P < 0. 05 ) . Except the 21st day point of the AN 1. 25 mg/kg group, the mean optical density values of Bax in all time points of AN exposure groups were significantly higher than the negative control group (P < 0. 05 ). The decreased expression of Bcl-2 protein was most distinct in AN 2. 50mg/kg group and the positive control group at all time points. The expression of Bax protein was significantly increased in all groups at the 14th day point. Conclusion The expression of Bel-2 protein could be weakened in spermatogenic cells induced by AN, especially in the AN 2. 50 mg/kg8roup; while the expression of Bax was enhanced, and the amplitude of change in the 14th day point was more obvious.%目的 探讨丙烯腈(AN)暴露对小鼠睾丸生精细胞Bcl-2、Bax蛋白表达水平的影响.方法 将250只成年健康SPF级昆明种雄性小鼠,按体重随机分为阴性对照组、3个AN染毒组和阳性对照组,阴性对照组用生理盐水,各染毒组分别以1.25、2.50、5.00mg/kg AN腹腔注射(注射剂量为0.01 ml/g BW),每天1次,连续5天,阳性

  7. Identification and expression profiles of genes and protens in SMMC-7721 cells.

    Science.gov (United States)

    Feng, Xie Shun; Ming, Sun Shu; Feng, Lu Xiao; Feng, Xie Ze

    2014-02-01

    In the study presented here, we first evaluated effect of CDDP on liver cancer cells SMMC-7721 apoptosis and motility capacity. Then, we evaluate inhibitory effect of CDDP on tumour growth and its possible molecular mechanism in liver cancer mice model. Results showed that the apoptosis rate of cells decreased with increasing CDDP. Analysis of the effect of the CDDP on cell cycle was performed by flow cytometry and results show a dose-dependent increase in the percentage of cells in the S-phase of the cell cycle, with a decrease in the percentage of cells in the G1 and G2/M phases. CDDP did not close the wound even after 48 h, as opposed to untreated cells (0 mg/l). Similarly, the migratory and invasion capacity of SMMC-7721 cells was also reduced after treatment with CDDP, as evaluated by a transwell assay. Animal experiment indicated that CDDP administration could increase blood WBC, total protein, albumin and A/G, decrease blood alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase levels in hepatocellular carcinomas mice. Immunohistochemistry analysis showed that positive expression of Fas and Bax proteins in the medicine-treated (II, III) group was significantly higher, whereas the expression of NF-κB, P53, Bcl-2 proteins was significantly lower than those of the control group. Gene expression analysis using Real time PCR methods revealed a significant up-regulation in the expression levels of Bax mRNA in the medicne-treated (II, III) group when compared to untreated control. In contrast, CDDP-treated group showed a significant down regulation in the expression levels of Bcl-2 mRNA as compared to untreated control group. These results are in agreement with immunohistochemistry data. Our observations indicate that CDDP has damaged effects on liver tumour cells SMMC-7721 including apoptosis, motility and cell cycle under in vitro. CDDP can enhance pro-apoptosis gene Fas, Bax expression, decrease anti-apoptosis genes Bcl-2 expression

  8. 特发性胎儿生长受限患者胎盘组织中XIAP和Bax的意义%Significances of XIAP and Bax in placental tissue of patients with idiopathic fetal growth restriction

    Institute of Scientific and Technical Information of China (English)

    崔世红; 张婷; 管秀娟; 程国梅; 贾国占; 刘萍萍; 杨健丽

    2012-01-01

    Objective; To explore the expressions and significances of XIAP and Bax in placental tissue of patients with idiopathic fetal growth restriction (IFGR) . Methods: Thirty pregnant women with IFGR who gave birth to their babies in the hospital via cesarean section from July 2010 to April 2011 were selected as experimental group, while thirty full - term pregnant women who gave birth to their babies in the hospital via cesarean section because of social factor during the same period were selected as control group. Immunohistochemical method was used to detect the expression levels of XIAP and Bax in placental tissue. Results: The expression level of XIAP in placental syn-cyliolrophoblast cells of experimental group was ( 114. 562 ± 5. 167) , which was significantly lower than that of control group (144. 430 ± 7. 311) (P 0. 05) . Conclusion: Down - regulation of XIAP and up - regulation of Bax in placental tissue can promote excessive apoptosis of placental syncytiotrophoblast cells, which may be one of the important pathogenesis of IFGR.%目的:探讨特发性胎儿生长受限(IFGR)胎盘组织中XIAP、Bax的表达及意义.方法:选取我院2010年7月~2011年4月剖宫产分娩的IFGR孕妇30例作为实验组,同期因社会因素剖宫产分娩的正常足月孕妇30例作为对照组.用免疫组化法检测胎盘组织XIAP、Bax的表达水平.结果:①XIAP在胎盘合体滋养细胞中的表达,实验组(114.562±5.167)低于对照组(144.430 +7.311),差异有统计学意义(P<0.05).②Bax在胎盘合体滋养细胞中的表达,实验组(146.513±10.611)高于对照组(113.673±9.631),差异有统计学意义(P<0.05);③实验组胎盘组织中XIAP与Bax的表达呈负相关(r=-0.761,P<0.05);对照组胎盘组织中XIAP与Bax的表达无相关性(r=0.034,P>0.05).结论:胎盘组织中XIAP表达降低、Bax表达增加促使了胎盘滋养细胞的过度凋亡,可能是IFGR发病机制中的重要环节之一.

  9. Gene therapy

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    2005147 CNHK200-hA-a gene-viral therapeutic system and its antitumor effect on lung cancer. WANG Wei-guo(王伟国),et al. Viral & Gene Ther Center, Eastern Hepatobilli Surg Instit 2nd Milit Univ, Shanghai 200438. Chin J Oncol,2005:27(2):69-72. Objective: To develop a novel vector system, which combines the advantages of the gene therapy,

  10. 视网膜母细胞瘤Bcl-2和Bax基因蛋白质表达%Expression of Bcl-2 and Bax gene protein in retinoblastoma

    Institute of Scientific and Technical Information of China (English)

    张小猛; 庞利民; 张晓光

    2000-01-01

    目的:研究凋亡及凋亡调控基因Bcl-2/Bax和视网膜母细胞瘤(Retinoblastoma,RB)的发生、发展及退化的关系.方法:收集36例RB标本,对其分别进行Bcl-2和Bax基因的蛋白质免疫组织化学染色.对其表达情况和染色强度进行观察.结果:①Bcl-2在分化型RB中表达比较好;②Bax在未分化型和分化型中表达都比较好.结论:①分化型和未分化型RB中都有Bcl-2/Bax基因蛋白表达;②随RB恶性度的增加,Bcl-2的表达逐渐减弱;Bax的表达无明显改变.③分化型RB受Bcl-2和Bax基因共同控制;未分化型RB受Bax基因调控,Bcl-2基因发挥很少的作用.

  11. Human Thyroid Cancer-1 (TC-1) is a vertebrate specific oncogenic protein that protects against copper and pro-apoptotic genes in yeast

    OpenAIRE

    Natalie K. Jones; Nagla T.T. Arab; Rawan Eid; Nada Gharib; Sara Sheiban; Hojatollah Vali; Chamel Khoury; Alistair Murray; Eric Boucher; Mandato, Craig A; Young, Paul G.; Greenwood, Michael T

    2015-01-01

    The human Thyroid Cancer-1 (hTC-1) protein, also known as C8orf4 was initially identified as a gene that was up-regulated in human thyroid cancer. Here we show that hTC-1 is a peptide that prevents the effects of over-expressing Bax in yeast. Analysis of the 106 residues of hTC-1 in available protein databases revealed direct orthologues in jawed-vertebrates, including mammals, frogs, fish and sharks. No TC-1 orthologue was detected in lower organisms, including yeast. Her...

  12. X-linked inhibitor of apoptosis regulates T cell effector function

    DEFF Research Database (Denmark)

    Zehntner, Simone P; Bourbonnière, Lyne; Moore, Craig S;

    2007-01-01

    To understand how the balance between pro- and anti-apoptotic signals influences effector function in the immune system, we studied the X-linked inhibitor of apoptosis (XIAP), an endogenous regulator of cellular apoptosis. Real-time PCR showed increased XIAP expression in blood of mice...... and oligodendrocytes were not affected; neither did apoptosis increase in liver, where XIAP knockdown also occurred. ASO-XIAP increased susceptibility of T cells to activation-induced apoptosis in vitro. Our results identify XIAP as a critical controller of apoptotic susceptibility of effector T cell function...

  13. Osteocyte apoptosis regulates osteoclast precursor adhesion via osteocytic IL-6 secretion and endothelial ICAM-1 expression.

    Science.gov (United States)

    Cheung, Wing-Yee; Simmons, Craig A; You, Lidan

    2012-01-01

    Osteocyte apoptosis precedes osteoclast resorption, and may act as a critical signal to trigger bone remodeling. While osteoclast precursors are known to travel via the circulation, the specific mechanisms by which they accumulate at remodeling sites are unclear. We hypothesized that osteocyte apoptosis mediates osteoclast precursor adhesion to vascular endothelium by regulating osteocytic secretion of IL-6 and soluble IL-6 receptor (sIL-6R) to promote endothelial ICAM-1 expression. We found that conditioned media from TNF-α-induced apoptotic MLO-Y4 osteocytes promoted RAW264.7 osteoclast precursor adhesion onto D4T endothelial cells (P<0.05). Blocking osteocyte apoptosis with a pan-caspase inhibitor (ZVAD-FMK) reduced osteoclast precursor adhesion to baseline levels (P<0.001). Endothelial cells treated with apoptotic osteocyte conditioned media had elevated surface expression of ICAM-1 (P<0.05), and blocking ICAM-1 abolished apoptosis-induced osteoclast precursor adhesion. Apoptotic osteocyte conditioned media contained more IL-6 (P<0.05) and sIL-6R (P<0.05) than non-apoptotic osteocyte conditioned media. When added exogenously, both IL-6 and sIL-6R were required for endothelial activation, and blocking IL-6 reduced apoptosis-induced osteoclast precursor adhesion to baseline levels (P<0.05). Therefore, we conclude that osteocyte apoptosis can promote osteoclast precursor adhesion to endothelial cells via ICAM-1; this is likely through increased osteocytic IL-6 and sIL-6R secretion, both of which are indispensible to endothelial activation. PMID:21986000

  14. Tissue microarrays in pathological examination of apoptotic acinar cells induced by dexamethasone in the pancreas of rats with severe acute pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Xi-Ping Zhang; Hua Tian; Bei Lu; Li Chen; Ru-Jun Xu; Ke-Yi Wang; Zhi-Wei Wang; Qi-Hui Cheng; Hai-Ping Shen

    2007-01-01

    BACKGROUND:The good therapeutic effects of large dose of dexamethasone on severe acute pancreatitis (SAP) patients have been proved. This study was designed to investigate the inlfuence of dexamethasone on apoptosis of acinar cells in the pancreas of rats with SAP and the protein expression of the apoptosis-regulating genes Bax and Bcl-2. METHODS: Ninety Sprague-Dawley rats with SAP were randomly divided into a model group and a dexamethasone treated group (45 rats in each group), and another 45 rats formed the sham operation group. Survival rates were calculated and gross pathological changes in the pancreas of each group were observed under a light microscope 3, 6 and 12 hours after operation. Tissue microarray technology was applied to prepare pancreatic tissue sections. The changes in Bax and Bcl-2 protein expression levels of pancreatic tissues from each group were assessed by immunohistochemical staining, and TUNEL staining was used to evaluate changes in apoptosis index. RESULTS: The model and treated groups did not differ in mortality at each time point. The pathological score for the pancreas in the treated group was signiifcantly lower than that in the model group at 3 and 6 hours. The positive rates of Bax protein expression in the head and tail of the pancreas in the treated group at all time points were all markedly higher than those of the model group. The positive rate of Bcl-2 protein expression in the head of the pancreas in the treated group was signiifcantly higher than that of the model group at 3 hours. TUNEL staining showed that the pancreas head and tail apoptosis indices of the treated group were markedly higher than those of the model group after 6 hours. CONCLUSIONS: Apoptosis may be a protective response to pancreatic cell injury. The mechanism of action of dexamethasone in treating SAP may be related to the apoptosis of acinar cells in the pancreas induced by apoptosis-regulating genes such as Bax and Bcl-2. The advantages of tissue

  15. Post operative infection and sepsis in humans is associated with deficient gene expression of gammac cytokines and their apoptosis mediators.

    LENUS (Irish Health Repository)

    White, Mary

    2011-06-28

    Abstract Introduction Lymphocyte homeostasis is dependent on the γc cytokines. We hypothesised that sepsis in humans is associated with differential gene expression of the γc cytokines and their associated apoptosis mediators. Methods The study population consisted of a total of 60 patients with severe sepsis, 15 with gram negative bacteraemia, 10 healthy controls and 60 patients undergoing elective lung resection surgery. Pneumonia was diagnosed by CDC NNIC criteria. Gene expression in peripheral blood leukocytes (PBLs) of interleukin (IL)-2, 7, 15 and interferon (IFN)-γ, Bax, Bim, Bcl-2 was determined by qRT-PCR and IL-2 and IL-7 serum protein levels by ELISA. Gene expression of IL-2, 7 and IFN-γ was measured in peripheral blood leukocytes (PBL), cultured in the presence of lipopolysacharide (LPS) and CD3 binding antibody (CD3ab) Results IL-2 gene expression was lower in the bacteraemia group compared with controls, and lower still in the sepsis group (P < 0.0001). IL-7 gene expression was similar in controls and bacteraemia, but lower in sepsis (P < 0.0001). IL-15 gene expression was similar in the three groups. Bcl-2 gene expression was less (P < 0.0001) and Bim gene expression was greater (P = 0.0003) in severe sepsis compared to bacteraemic and healthy controls. Bax gene expression was similar in the three groups. In lung resection surgery patients, post-operative pneumonia was associated with a perioperative decrease in IL-2 mRNA (P < 0.0001) and IL-7 mRNA (P = 0.003). IL-2 protein levels were reduced in sepsis and bacteraemia compared to controls (P = 0.02) but similar in pneumonia and non-pneumonia groups. IL-7 protein levels were similar in all groups. In cultured PBLs, IFN-γ gene expression was decreased in response to LPS and increased in response to CD3ab with sepsis: IL-7 gene expression increased in response to LPS in controls and to CD3ab with sepsis; Bcl-2 gene expression decreased in response to combined CD3ab and IL-2 with sepsis

  16. Dopaminergic neuron-specific deletion of p53 gene is neuroprotective in an experimental Parkinson's disease model.

    Science.gov (United States)

    Qi, Xin; Davis, Brandon; Chiang, Yung-Hsiao; Filichia, Emily; Barnett, Austin; Greig, Nigel H; Hoffer, Barry; Luo, Yu

    2016-09-01

    p53, a stress response gene, is involved in diverse cell death pathways and its activation has been implicated in the pathogenesis of Parkinson's disease (PD). However, whether the neuronal p53 protein plays a direct role in regulating dopaminergic (DA) neuronal cell death is unknown. In this study, in contrast to the global inhibition of p53 function by pharmacological inhibitors and in traditional p53 knock-out (KO) mice, we examined the effect of DA specific p53 gene deletion in DAT-p53KO mice. These DAT-p53KO mice did not exhibit apparent changes in the general structure and neuronal density of DA neurons during late development and in aging. However, in DA-p53KO mice treated with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), we found that the induction of Bax and p53 up-regulated modulator of apoptosis (PUMA) mRNA and protein levels by MPTP were diminished in both striatum and substantia nigra of these mice. Notably, deletion of the p53 gene in DA neurons significantly reduced dopaminergic neuronal loss in substantia nigra, dopaminergic neuronal terminal loss at striatum and, additionally, decreased motor deficits in mice challenged with MPTP. In contrast, there was no difference in astrogliosis between WT and DAT-p53KO mice in response to MPTP treatment. These findings demonstrate a specific contribution of p53 activation in DA neuronal cell death by MPTP challenge. Our results further support the role of programmed cell death mediated by p53 in this animal model of PD and identify Bax, BAD and PUMA genes as downstream targets of p53 in modulating DA neuronal death in the in vivo MPTP-induced PD model. We deleted p53 gene in dopaminergic neurons in late developmental stages and found that DA specific p53 deletion is protective in acute MPTP animal model possibly through blocking MPTP-induced BAX and PUMA up-regulation. Astrocyte activation measured by GFAP positive cells and GFAP gene up-regulation in the striatum shows no difference

  17. Effect of Sargassum fusiforme Polysaccharide on the Ethology and Expressions of Bcl-2 and Bax in Brain Tissue for Alzheimer's Disease Rat model%羊栖菜多糖对老年痴呆模型大鼠Bcl-2和Bax基因表达的分析

    Institute of Scientific and Technical Information of China (English)

    汤从容; 曹高忠; 叶晓兰

    2012-01-01

    Objective:To study effect of Sargassum fusiforme Polysaccharide( SFPS) on ethology and expressions of Bcl - 2 and Bax in brain tissue of AD rats. Methods:The D - galactose AD rat model was applied,and blank group,model group, Piracetam control group and herbal group were designed. Index change of ethology , expressions of Bcl -2 and Bax in brain tissue were tested. Results: AD model may decrease cognitive ability,the ratio of Bcl -2/Bax in the hippocampus came down(P <0.05 ). Compared to the model group,Sargassum fusiforme Polysaccharide with 0. 8 ,1. 6g/kg can alleviate cognitive ability significantly,and the action had a does —dependent increase,the ratio of Bcl -2/Bax increased(P< 0.05). Conclusion; Sargassum fusiforme Polysaccharide can up - regulate expression of protein Bax and down -regulate expression of Bcl - 2 in brain tissu. It inhibits apoptosis through regulating the expressions of Bcl - 2 and Bax protein in hippocampal , which might be one of mechanisms of Sargassum fusiforme Polysaccharide to prevent and treat AD disease.%目的:探讨羊栖菜多糖提取物(SFPS)对阿尔茨海默病(AD)大鼠模型行为的干预作用及脑皮质Bcl-2和Bax基因表达的影响.方法:制作D-半乳糖阿尔茨海默病大鼠模型,设计正常对照组、模型组、吡拉西坦片、羊栖菜多糖提取物不同剂量组,观察大鼠行为学及脑皮质Bcl -2和Bax基因表达指标的改变.结果:与正常对照组相比,模型组学习记忆能力显著下降(P<0.05),其Bcl - 2/Bax值下降;与模型组相比,0.8g/kg、1.6g/kg羊栖菜多糖提取物治疗组均能较好的改善学习记忆能力,且具有一定剂量依赖性,其Bcl - 2/Bax值增加.结论:SFPS能调节海马组织Bcl -2和Bax的表达,显著提高Bcl - 2/Bax值,抑制海马神经元的凋亡,改善AD大鼠学习记忆能力.

  18. Bisphenol A induces spermatocyte apoptosis in rare minnow Gobiocypris rarus.

    Science.gov (United States)

    Zhang, Yingying; Cheng, Mengqian; Wu, Lang; Zhang, Guo; Wang, Zaizhao

    2016-10-01

    Bisphenol A (BPA) is an endocrine disruptor, and could induce germ cells apoptosis in the testis of mammals. But whether it could affect fish in the same mechanism has not' been studied till now. In the present study, to investigate the influence of BPA on testis germ cells in fish, adult male rare minnow Gobiocypris rarus were exposed to 225μgL(-1) (0.99μM) BPA for 1, 3 and 9 weeks. Through TdT-mediated dUTP nick end labeling (TUNEL) and transmission electron microscope (TEM) analysis, we found that the amount of apoptotic spermatocytes significantly increased in a time dependent manner following BPA exposure. Western Blot results showed that the ratio of Bcl2/Bax, the important apoptosis regulators in intrinsic mitochondrial apoptotic pathway, was significantly decreased. qPCR showed that mRNA expression of several genes in mitochondrial apoptotic pathway including bcl2, bax, casp9, cytc and mcl1b were significantly changed following BPA exposure. In addition, mRNA expression of meiosis regulation genes (kpna7 and wee2), and genes involved in both apoptosis and meiosis (birc5, ccna1, and gsa1a) were also affected by BPA. Taken together, the present study demonstrated that BPA could induce spermatocytes apoptosis in rare minnow testis, and the apoptosis was probably under regulation of intrinsic mitochondrial apoptotic pathway. Moreover, the spermatocyte apoptosis was likely initiated by BPA induced meiosis arrest. PMID:27561114

  19. Trichoderma genes

    Science.gov (United States)

    Foreman, Pamela; Goedegebuur, Frits; Van Solingen, Pieter; Ward, Michael

    2012-06-19

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

  20. Gene therapy.

    OpenAIRE

    Mota Biosca, Anna

    1992-01-01

    Applications of gene therapy have been evaluated in virtually every oral tissue, and many of these have proved successful at least in animal models. While gene therapy will not be used routinely in the next decade, practitioners of oral medicine should be aware of the potential of this novel type of treatment that doubtless will benefit many patients with oral diseases.

  1. Expressions of transcription factors Smad4 and NF-κB in preeclampsia placenta tissue and exploration of its relationship with expressions of apoptotic and invasive genes

    Institute of Scientific and Technical Information of China (English)

    Li Xia

    2015-01-01

    Objective:To study the expressions of transcription factors Smad4 and NF-κB in preeclampsia placenta tissue and its relationship with expressions of apoptotic and invasive genes.Methods:50 cases of preeclampsia puerperal women and 50 cases of normal puerperal women treated and gave birth in our hospital from May 2012 to May 2014 were chosen for study. Placenta tissue was collected and PCR method was used to detect mRNA contents of Smad4, NF-κB, Fas, FasL, Caspase-3, Caspase-8, Bax, MMP2, MMP9, IL-24 and RECK; immunohistochemical method was used to detect positive expressions of Smad4 and NF-κB.Results: Compared with normal placenta tissue, mRNA contents and immunohistochemical positive staining rates of Smad4 and NF-κB in preeclampsia placenta were all higher; contents of Fas, FasL, Caspase-3, Caspase-8, Bax, IL-24 and RECK of Smad-positive group and NF-κB-positive group were higher than those of Smad-negative group and NF-κB-negative group respectively; MMP2 and MMP9 contents were lower than those of Smad-negative group and NF-κB-negative group respectively.Conclusion: Smad4 and NF-κB expressions in preeclampsia placenta abnormally increase and may regulate the expressions of apoptotic genes and invasive genes to be involved in the occurrence of the disease.

  2. A-site compositional effects in Ga-doped hollandite materials of the form BaxCsyGa2x+yTi8−2x−yO16: implications for Cs immobilization in crystalline ceramic waste forms

    Science.gov (United States)

    Xu, Yun; Wen, Yi; Grote, Rob; Amoroso, Jake; Shuller Nickles, Lindsay; Brinkman, Kyle S.

    2016-01-01

    The hollandite structure is a promising crystalline host for Cs immobilization. A series of Ga-doped hollandite BaxCsyGa2x+yTi8−2x−yO16 (x = 0, 0.667, 1.04, 1.33; y = 1.33, 0.667, 0.24, 0) was synthesized through a solid oxide reaction method resulting in a tetragonal hollandite structure (space group I4/m). The lattice parameter associated with the tunnel dimension was found to increases as Cs substitution in the tunnel increased. A direct investigation of cation mobility in tunnels using electrochemical impedance spectroscopy was conducted to evaluate the ability of the hollandite structure to immobilize cations over a wide compositional range. Hollandite with the largest tunnel size and highest aspect ratio grain morphology resulting in rod-like microstructural features exhibited the highest ionic conductivity. The results indicate that grain size and optimized Cs stoichiometry control cation motion and by extension, the propensity for Cs release from hollandite. PMID:27273791

  3. A-site compositional effects in Ga-doped hollandite materials of the form BaxCsyGa2x+yTi8‑2x‑yO16: implications for Cs immobilization in crystalline ceramic waste forms

    Science.gov (United States)

    Xu, Yun; Wen, Yi; Grote, Rob; Amoroso, Jake; Shuller Nickles, Lindsay; Brinkman, Kyle S.

    2016-06-01

    The hollandite structure is a promising crystalline host for Cs immobilization. A series of Ga-doped hollandite BaxCsyGa2x+yTi8‑2x‑yO16 (x = 0, 0.667, 1.04, 1.33; y = 1.33, 0.667, 0.24, 0) was synthesized through a solid oxide reaction method resulting in a tetragonal hollandite structure (space group I4/m). The lattice parameter associated with the tunnel dimension was found to increases as Cs substitution in the tunnel increased. A direct investigation of cation mobility in tunnels using electrochemical impedance spectroscopy was conducted to evaluate the ability of the hollandite structure to immobilize cations over a wide compositional range. Hollandite with the largest tunnel size and highest aspect ratio grain morphology resulting in rod-like microstructural features exhibited the highest ionic conductivity. The results indicate that grain size and optimized Cs stoichiometry control cation motion and by extension, the propensity for Cs release from hollandite.

  4. A-site compositional effects in Ga-doped hollandite materials of the form BaxCsyGa2x+yTi8-2x-yO16: implications for Cs immobilization in crystalline ceramic waste forms.

    Science.gov (United States)

    Xu, Yun; Wen, Yi; Grote, Rob; Amoroso, Jake; Shuller Nickles, Lindsay; Brinkman, Kyle S

    2016-01-01

    The hollandite structure is a promising crystalline host for Cs immobilization. A series of Ga-doped hollandite BaxCsyGa2x+yTi8-2x-yO16 (x = 0, 0.667, 1.04, 1.33; y = 1.33, 0.667, 0.24, 0) was synthesized through a solid oxide reaction method resulting in a tetragonal hollandite structure (space group I4/m). The lattice parameter associated with the tunnel dimension was found to increases as Cs substitution in the tunnel increased. A direct investigation of cation mobility in tunnels using electrochemical impedance spectroscopy was conducted to evaluate the ability of the hollandite structure to immobilize cations over a wide compositional range. Hollandite with the largest tunnel size and highest aspect ratio grain morphology resulting in rod-like microstructural features exhibited the highest ionic conductivity. The results indicate that grain size and optimized Cs stoichiometry control cation motion and by extension, the propensity for Cs release from hollandite. PMID:27273791

  5. Triphala Extract Suppresses Proliferation and Induces Apoptosis in Human Colon Cancer Stem Cells via Suppressing c-Myc/Cyclin D1 and Elevation of Bax/Bcl-2 Ratio

    Directory of Open Access Journals (Sweden)

    Ramakrishna Vadde

    2015-01-01

    Full Text Available Colon cancer is the second leading cause of cancer related deaths in the USA. Cancer stem cells (CSCs have the ability to drive continued expansion of the population of malignant cells. Therefore, strategies that target CSCs could be effective against colon cancer and in reducing the risk of relapse and metastasis. In this study, we evaluated the antiproliferative and proapoptotic effects of triphala, a widely used formulation in Indian traditional medicine, on HCT116 colon cancer cells and human colon cancer stem cells (HCCSCs. The total phenolic content, antioxidant activity, and phytochemical composition (LC-MS-MS of methanol extract of triphala (MET were also measured. We observed that MET contains a variety of phenolics including naringin, quercetin, homoorientin, and isorhamnetin. MET suppressed proliferation independent of p53 status in HCT116 and in HCCSCs. MET also induced p53-independent apoptosis in HCCSCs as indicated by elevated levels of cleaved PARP. Western blotting data suggested that MET suppressed protein levels of c-Myc and cyclin D1, key proteins involved in proliferation, and induced apoptosis through elevation of Bax/Bcl-2 ratio. Furthermore, MET inhibited HCCSCs colony formation, a measure of CSCs self-renewal ability. Anticancer effects of triphala observed in our study warrant future studies to determine its efficacy in vivo.

  6. Preparation of nanostructured La0.7Ca0.3-xBaxMnO3 ceramics by a combined sol-gel and spark plasma sintering route and resulting magnetocaloric properties

    Science.gov (United States)

    Ayadi, F.; Regaieg, Y.; Cheikhrouhou-Koubaa, W.; Koubaa, M.; Cheikhrouhou, A.; Lecoq, H.; Nowak, S.; Ammar, S.; Sicard, L.

    2015-05-01

    This work proposes an original, easy to achieve and inexpensive route to synthesize manganite ceramics for magnetic refrigeration, combining sol-gel chemistry to Spark Plasma Sintering (SPS). The target La0.7Ca0.3-xBaxMnO3 (x=0, 0.1, 0.2) compounds are obtained as single phases which crystallize in the orthorhombic structure (Pnma space group). SPS allows a quick sintering at a relatively low temperature (700 °C in this work) compared to the conventional solid state method (≥1100 °C), leading to densified ultrafine grained pellets (85% of compactness). Magnetic studies show that Ba substitution does not affect significantly the relative cooling power (RCP) of these manganites, while it increases their Curie temperature (TC) by several tens of degrees. Typically, RCP values ranging between 267 and 270 J/kg (for a magnetic field change of 5 T) and TC between 205 and 245 K were measured when x was increased from 0 to 0.2, respectively. These results combined to the fact that the synthesis route is economically advantageous makes the obtained ceramics interesting as active refrigerants for magnetic refrigeration technology below room temperature.

  7. Effect of Heroin Dependence on Expression of Bcl-2 and Bax in Submandibular Gland of Rats%海洛因依赖对大鼠颌下腺Bcl-2和Bax表达的影响

    Institute of Scientific and Technical Information of China (English)

    胡赟; 梁文妹; 洪艳; 韩晶; 夏白娟; 李一欣

    2013-01-01

    目的:观察海洛因依赖对大鼠颌下腺Bcl-2、Bax阳性细胞表达及分布的影响.方法:正常SD大鼠,随机分为正常对照组和海洛因依赖组,皮下注射海洛因建立海洛因依赖大鼠模型,分别于第10、17、24、31、38天取颌下腺组织,用免疫组织化学Envision法并结合图像分析方法,观察Bcl-和Bax表达.结果:(1) Bcl-2阳性细胞数量在海洛因依赖第10天明显下降(P<0.05),此后开始增加,至第24天时达最高峰(P<0.05),而在第38天时低于正常水平(P<0.05);平均灰度值在依赖第10天明显上升(P<0.05),后开始下降,第24天时达最低值(P<0.05),至第31 ~38天时接近正常水平(P>0.05);(2)Bax阳性细胞数量在依赖第10天明显降低(P<0.05),第17天时又增加达峰值(P<0.05);之后即开始下降,第31-38天时接近正常水平(P>0.05);其平均灰度值在依赖第10天显著上升(P<0.05),而在第17天时达最低值(P<0.05),后随依赖时间延长,与正常水平无显著性差异(P>0.05).结论:海洛因对颌下腺Bcl-2、Bax的表达有明显影响,且与依赖时间长短有关.%Objective:To observe the effect of heroin dependence on Bcl-2 and Bax positive cells in the submandibular gland of rats.Methods:Normal SD rats were divided into normal control group (NGC) and heroin dependence group (HDG).The heroin dependent model of rats was established by subcutaneous injection of heroin,and submandibular glands were excised on the following 10th,17th,24th,31st and 38th days respectively.Immunohistochemical Envision method and image analysis were used in the study.Results:1.The number of Bcl-2 positive cells descended obviously on the 10th d (P < 0.05),then,increased and peaked on the 24th d (P < 0.05).However,it was lower than that of NCG on the 38th d (P < 0.05) ; the mean grey value increased obviously on the 10th d (P < 0.05),but decreased and to the lowest on the 24th d (P <0.05).Between the 31st d and the 38th d

  8. Persea declinata (Bl. Kosterm Bark Crude Extract Induces Apoptosis in MCF-7 Cells via G0/G1 Cell Cycle Arrest, Bcl-2/Bax/Bcl-xl Signaling Pathways, and ROS Generation

    Directory of Open Access Journals (Sweden)

    Putri Narrima

    2014-01-01

    Full Text Available Persea declinata (Bl. Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill, which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl. Kosterm bark methanolic crude extract (PDM. PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48 h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.

  9. Persea declinata (Bl.) Kosterm Bark Crude Extract Induces Apoptosis in MCF-7 Cells via G0/G1 Cell Cycle Arrest, Bcl-2/Bax/Bcl-xl Signaling Pathways, and ROS Generation.

    Science.gov (United States)

    Narrima, Putri; Paydar, Mohammadjavad; Looi, Chung Yeng; Wong, Yi Li; Taha, Hairin; Wong, Won Fen; Ali Mohd, Mustafa; Hadi, A Hamid A

    2014-01-01

    Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48 h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development. PMID:24808916

  10. The Magea gene cluster regulates male germ cell apoptosis without affecting the fertility in mice

    Science.gov (United States)

    Hou, Siyuan; Xian, Li; Shi, Peiliang; Li, Chaojun; Lin, Zhaoyu; Gao, Xiang

    2016-01-01

    While apoptosis is essential for male germ cell development, improper activation of apoptosis in the testis can affect spermatogenesis and cause reproduction defects. Members of the MAGE-A (melanoma antigen family A) gene family are frequently clustered in mammalian genomes and are exclusively expressed in the testes of normal animals but abnormally activated in a wide variety of cancers. We investigated the potential roles of these genes in spermatogenesis by generating a mouse model with a 210-kb genomic deletion encompassing six members of the Magea gene cluster (Magea1, Magea2, Magea3, Magea5, Magea6 and Magea8). Male mice carrying the deletion displayed smaller testes from 2 months old with a marked increase in apoptotic germ cells in the first wave of spermatogenesis. Furthermore, we found that Magea genes prevented stress-induced spermatogenic apoptosis after N-ethyl-N-nitrosourea (ENU) treatment during the adult stage. Mechanistically, deletion of the Magea gene cluster resulted in a dramatic increase in apoptotic germ cells, predominantly spermatocytes, with activation of p53 and induction of Bax in the testes. These observations demonstrate that the Magea genes are crucial in maintaining normal testicular size and protecting germ cells from excessive apoptosis under genotoxic stress. PMID:27226137

  11. The Magea gene cluster regulates male germ cell apoptosis without affecting the fertility in mice.

    Science.gov (United States)

    Hou, Siyuan; Xian, Li; Shi, Peiliang; Li, Chaojun; Lin, Zhaoyu; Gao, Xiang

    2016-01-01

    While apoptosis is essential for male germ cell development, improper activation of apoptosis in the testis can affect spermatogenesis and cause reproduction defects. Members of the MAGE-A (melanoma antigen family A) gene family are frequently clustered in mammalian genomes and are exclusively expressed in the testes of normal animals but abnormally activated in a wide variety of cancers. We investigated the potential roles of these genes in spermatogenesis by generating a mouse model with a 210-kb genomic deletion encompassing six members of the Magea gene cluster (Magea1, Magea2, Magea3, Magea5, Magea6 and Magea8). Male mice carrying the deletion displayed smaller testes from 2 months old with a marked increase in apoptotic germ cells in the first wave of spermatogenesis. Furthermore, we found that Magea genes prevented stress-induced spermatogenic apoptosis after N-ethyl-N-nitrosourea (ENU) treatment during the adult stage. Mechanistically, deletion of the Magea gene cluster resulted in a dramatic increase in apoptotic germ cells, predominantly spermatocytes, with activation of p53 and induction of Bax in the testes. These observations demonstrate that the Magea genes are crucial in maintaining normal testicular size and protecting germ cells from excessive apoptosis under genotoxic stress. PMID:27226137

  12. 甘草黄酮对大强度耐力运动大鼠肾脏组织Ca2+、Mg2+-ATPase及Bax,Bc1-2蛋白表达的影响%Effect of High-Intensity Endurance Exercise on Ca2+,Mg2+-ATPase and Bax, Bcl-2 Protein Expression With Glycyrrhiza Flavonoids in rat Nephridial Tissue

    Institute of Scientific and Technical Information of China (English)

    王东旭; 陈艳艳

    2013-01-01

    Objective To explore Glycyrrhiza Elavonoids on the rat nephridial tissue of Ca2+, Mg2+-ATPase and Bax, Bcl-2 protein expression with high-intensity endurance exercise. Methods The twenty-four healthy male rats were randomly divided into quiet groups, high-intensity exercise group and exercise plus Glycyrrhiza Elavonoids group, After 6 weeks of treadmill training, Using the box of reagent and immunity histochemistry examined the changing of Ca2+, Mg2+-ATPase and Bax, Bcl-2 protein expression on each groups . Results Compared with the quiet groups, the activity of Ca2+, Mg2+-ATPase both had significant droped (P<0.01), and the groups of plus drog had very difference increased than high-intendity exerxise groups (P<0.01); High-intensity endurance exercise group and exercise dosing rats AI apoptosis index increased in varying degrees;high-intensity exercise group (MOD) were very significant difference(P<0.01), exercise plus drug group Bac protein expression (MOD)were very significant difference (P<0.01); Exercise plus drug group Bcl-2 protein expression(MOD) with the high-intersity exercise group had significant difference(P<0.01), High-intensity exercise group and exercise plus drug group Bax/Bcl-2 ratio of distribution is significantly difference degrees of difference(P<0.05,P<0.01).%目的:探讨甘草黄酮对大强度耐力运动大鼠肾脏组织Ca2+、Mg2+-ATPase及Bax、Bcl-2表达的影响。方法:选取SD雄性健康大鼠24只,随机分为安静组、大强度运动组和运动加药组;采用跑台训练6周后取材,应用试剂盒和免疫组织化学法测检测各组大鼠肾脏组织Ca2+、Mg2+-TPase活性及Bax和Bcl-2表达的变化。结果:与安静对照组相比,大强度运动组和运动加药组肾脏组织Ca2+、Mg2+-TPase活性均呈非常显著性下降(P<0.01);其中运动加药组Ca2+、Mg2+-TPase活性均较大强度运动组具有非常显著差异性提高(P<0.01);大强度耐力运动组和运动加

  13. Anticancer activity of resveratrol on implanted human primary gastric carcinoma cells in nude mice

    Institute of Scientific and Technical Information of China (English)

    Hai-Bo Zhou; Juan-Juan Chen; Wen-Xia Wang; Jian-Ting Cai; Qin Du

    2005-01-01

    AIM: To investigate the apoptosis of implanted primary gastric cancer cells in nude mice induced by resveratrol and the relation between this apoptosis and expression of bcl-2and bax.METHODS: A transplanted tumor model was established by injecting human primary gastric cancer cells into subcutaneous tissue of nude mice. Resveratrol (500 mg/kg, 1000 mg/kg and 1500 mg/kg) was directly injected beside tumor body 6 times at an interval of 2 d. Then changes of tumor volume were measured continuously and tumor inhibition rate of each group was calculated. We observed the morphologic alterations by electron microscope, measured the apoptotic rate by TUNEL staining method, detected the expression of apoptosis-regulated genes bcl-2and bax by immunohistochemical staining and PT-PCR.RESULTS: Resveratrol could significantly inhibit carcinoma growth when it was injected near the carcinoma. An inhibitory effect was observed in all therapeutic groups and the inhibition rate of resveratrol at the dose of 500 mg/kg,1 000 mg/kg and 1 500 mg/kg was 10.58%, 29.68% and 39.14%, respectively. Resveratrol induced implanted tumor cells to undergo apoptosis with apoptotic characteristics,including morphological changes of chromatin condensation,chromatin crescent formation, nucleus fragmentation. The inhibition rate of 0.2 mL of normal saline solution, 1 500 mg/kg DMSO, 500 mg/kg resveratrol, 1 000 mg/kg resveratrol, and 1 500 mg/kg resveratrol was L3.68±0.37%, 13.8±0.43%,48.7±1.07%, 56.44±1.39% and 67±0.96%, respectively. The positive rate of bcl-2 protein of each group was 29.48±0.51%,27.56±1.40%, 11.86±0.97%, 5.7±0.84% and 3.92±0.85%,respectively by immunohistochemical staining. The positive rate of bax protein of each group was 19.34±0.35%,20.88±0.91%, 40.02±1.20%, 45.72±0.88% and 52.3±1.54%,respectively by immunohistochemical staining. The density of bcl-2 mRNA in 0.2 mL normal saline solution, 1 500 mg/kg DMSO, 500 mg/kg resveratrol, 1 000 mg/kg resveratrol,and 1 500 mg

  14. α-Phellandrene alters expression of genes associated with DNA damage, cell cycle, and apoptosis in murine leukemia WEHI-3 cells.

    Science.gov (United States)

    Lin, Jen-Jyh; Yu, Chien-Chih; Lu, Kung-Wen; Chang, Shu-Jen; Yu, Fu-Shun; Liao, Ching-Lung; Lin, Jaung-Geng; Chung, Jing-Gung

    2014-08-01

    α-phellandrene (α-PA) is a cyclic monoterpene, present in natural plants such as Schinus molle L. α-PA promotes immune responses in mice in vivo. However, there is no available information on whether α-PA affects gene expression in leukemia cells. The present study determined effects of α-PA on expression levels of genes associated with DNA damage, cell cycle and apoptotic cell death in mouse leukemia WEHI-3 cells. WEHI-3 cells were treated with 10 μM α-PA for 24 h, cells were harvested and total RNA was extracted, and gene expression was analyzed by cDNA microarray. Results indicated that α-PA up-regulated 10 genes 4-fold, 13 by over 3-fold and 175 by over 2-fold; 21 genes were down-regulated by over 4-fold, 26 genes by over 3-fold and expression of 204 genes was altered by at leas 2-fold compared with the untreated control cells. DNA damage-associated genes such as DNA damage-inducer transcript 4 and DNA fragmentation factor were up-regulated by 4-fold and over 2-fold, respectively; cell-cycle check point genes such as cyclin G2 and cyclin-dependent kinases inhibitor 2D and IA (p21) were up-regulated by over 3-fold and over 2-fold, respectively; apoptosis-associated genes such as BCL2/adenovirus EIB interacting protein 3, XIAP-associated factor 1, BCL2 modifying factor, caspase-8 and FADD-like apoptosis regulator were over 2-fold up-regulated. Furthermore, DNA damage-associated gene TATA box binding protein was over 4-fold down-regulated, and D19Ertd652c (DNA segment) over 2-fold down-regulated; cell cycle-associated gene cyclin E2 was over 2-fold down-regulated; apoptosis associated gene growth arrest-specific 5 was over 9-fold down-regulated, Gm5426 (ATP synthase) was over 3-fold down-regulated, and death box polypeptide 33 was over 2-fold down-regulated. Based on these observations, α-PA altered gene expression in WEHI-3 cells in vitro. PMID:25075043

  15. [Language gene].

    Science.gov (United States)

    Takahashi, Hiroshi

    2006-11-01

    The human capacity for acquiring speech and language must derive, at least in part, from the genome. Recent advance in the field of molecular genetics finally discovered 'Language Gene'. Disruption of FOXP2 gene, the firstly identified 'language gene' causes severe speech and language disorder. To elucidate the anatomical basis of language processing in the brain, we examined the expression pattern of FOXP2/Foxp2 genes in the monkey and rat brains through development. We found the preferential expression of FOXP2/Foxp2 in the striosomal compartment of the developing striatum. Thus, we suggest the striatum, particularly striosomal system may participate in neural information processing for language and speech. Our suggestion is consistent with the declarative/ procedural model of language proposed by Ullman (1997, 2001), which the procedural memory-dependent mental grammar is rooted in the basal ganglia and the frontal cortex, and the declarative memory-dependent mental lexicon is rooted in the temporal lobe. PMID:17432197

  16. Heat shock genes – integrating cell survival and death

    Indian Academy of Sciences (India)

    Richa Arya; Moushami Mallik; Subhash C Lakhotia

    2007-04-01

    Heat shock induced gene expression and other cellular responses help limit the damage caused by stress and thus facilitate cellular recovery. Cellular damage also triggers apoptotic cell death through several pathways. This paper briefly reviews interactions of the major heat shock proteins with components of the apoptotic pathways. Hsp90, which acts as a chaperone for unstable signal transducers to keep them poised for activation, interacts with RIP and Akt and promotes NF-B mediated inhibition of apoptosis; in addition it also blocks some steps in the apoptotic pathways. Hsp70 is mostly anti-apoptotic and acts at several levels like inhibition of translocation of Bax into mitochondria, release of cytochrome c from mitochondria, formation of apoptosome and inhibition of activation of initiator caspases. Hsp70 also modulates JNK, NF-B and Akt signaling pathways in the apoptotic cascade. In contrast, Hsp60 has both anti- and pro-apoptotic roles. Cytosolic Hsp60 prevents translocation of the pro-apoptotic protein Bax into mitochondria and thus promotes cell survival but it also promotes maturation of procaspase-3, essential for caspase mediated cell death. Our recent in vivo studies show that RNAi for the Hsp60D in Drosophila melanogaster prevents induced apoptosis. Hsp27 exerts its anti-apoptotic influence by inhibiting cytochrome c and TNF-mediated cell death. crystallin suppresses caspase-8 and cytochrome c mediated activation of caspase-3. Studies in our laboratory also reveal that absence or reduced levels of the developmentally active as well as stress induced non-coding hsr transcripts, which are known to sequester diverse hnRNPs and related nuclear RNA-binding proteins, block induced apoptosis in Drosophila. Modulation of the apoptotic pathways by Hsps reflects their roles as ``weak links” between various ``hubs” in cellular networks. On the other hand, non-coding RNAs, by virtue of their potential to bind with multiple proteins, can act as ``hubs” in

  17. Apoptosis of Hepatoma Cell Line HepG2 Induced by the Combination of Radiotherapy and Thermotherapy and Its Relationship with Bcl-2/Bax Protein Expressions%放疗联合热疗诱导肝癌HepG2细胞凋亡及其与Bcl-2和Bax蛋白表达关系的研究

    Institute of Scientific and Technical Information of China (English)

    张力; 龚明玉; 李毅学; 张立广; 王兴艳

    2011-01-01

    Objective To explore the apoptosis of hepatoma cell line HepG2 induced by the combination of radiotherapy and thermotherapy and its relationship with Bcl - 2/Bax protein expressions. Methods In vitro cultured HepG2 cells were randomly divided into four groups: control group ( not treated ), radiotherapy group, thermotherapy group, and combination group. The apoptosis of HepG2 cells were detected by flow cytometry. The expressions of the apoptosis-related proteins of Bcl-2 and Bax were detected by immunohistochemical methods. Results The apoptosis rates of HepG2 cells were significantly different among these four groups ( P < 0. 05 ). The apoptosis rates were significantly higher in radiotherapy group, thermotherapy group, and combination group than in control group ( P <0. 05 ). It was also significantly higher in combination group than in radiotherapy group and thermotherapy group ( P < 0. 05 ). The expressions of Bcl-2 and Bax and the Bax/Bcl-2 ratio were also significantly different among these four groups ( P <0. 05 ). The expression of Bcl -2 protein were significantly decreased and the expression of Bax protein significantly increased in radiotherapy group, thermotherapy group, and combination group than in control group ( both P < 0. 05 ), and the Bax/Bcl - 2 ratio was also significantly increased ( P < 0. 05 ). The expression of Bcl - 2 protein were significantly decreased and the expression of Bax protein significantly increased in combination group than in radiotherapy group and thermotherapy group ( both P < 0. 05 ), and the Bax/Bcl - 2 ratio was also significantly increased ( P < 0. 05 ). Conclusion The combination of radiotherapy and thermotherapy can more effectively induce the apoptosis of HepG2, and it may be achieved by inhibiting the expression of Bcl - 2 protein and promoting the expression of Bax protein.%目的 探讨放疗联合热疗诱导人肝癌HepG2细胞凋亡及其与Bcl-2和Bax蛋白表达的关系.方法

  18. 低频电磁场对大鼠生精细胞凋亡基因表达影响%Effect of extremely low frequency electromagnetic fields on Bax and Bcl-2 expression in spermatogenic cells of rats

    Institute of Scientific and Technical Information of China (English)

    孙晓芳; 段斐; 寇素茹; 刘晋芝; 马幼敏

    2009-01-01

    目的 探讨极低频电磁场对大鼠生精细胞凋亡相关基因Bax和Bcl-2表达的影响.方法 Wistar大鼠60只随机分为低剂量(0.1 mT)、高剂量(12.8 mT)电磁场暴露组及对照组,每组20只,分别有10只持续暴露2和12周;用苏木苏-伊红(H-E)染色及免疫组织化学法观察睾丸形态学和细胞凋亡相关基因Bax、Bcl-2表达的变化,用图像分析系统进行测定.结果 低、高剂量电磁场暴露组暴露2周Bax灰度值分别为(25.58±4.98),(39.17±4.33);Bcl-2灰度值分别为(68.12±3.64),(39.62±4.35);暴露12周Bax灰度值分别为(40.12±2.45),(62.32±5.90);Bcl-2灰度值分别为(55.78±2.43)(23.84±3.62).结论 随着电磁场暴露频率的升高和时间的延长,大鼠生精细胞Bax的表达明显上调,Bcl-2的表达逐渐下调.%Objective To study the effect of extremely low frequency electromagnetic fields on Boa and Bcl-2 expres-sion in the spermatogertic cells of rats. Methods Sixty adult female Wistar rats were divided into 0. 1 naT, 12. 8 rat and control group. After electromagnetic radiation for 2 and 12 weeks, HE staining and immunohistochemical methods were used to detected Bax and Bcl-2 expression with image analysis. Results After electromagnetic exposure for 2 week,the gray scale values of Bax were 25.58±4. 98 and 39. 17±4. 33 for low and high intensity groups and that of Bcl-2 were 68. 12± 3. 64 and 39. 62±4.35. The gray scale values of Boa were 40.12±2.45 and 62.32±5.90 for low and high intensity groups for 12 weeks exposure and that of Bcl-2 were 55.7±2.43 and 23.84±3.62. Conclusion With the increase of exposure in-tensity and time, the Box and Bcl-2 exprseeion in spermatogenie cells in rats were gradually decreased.

  19. High expression of the circadian gene mPer2 diminishes the radiosensitivity of NIH 3T3 cells

    Energy Technology Data Exchange (ETDEWEB)

    Chang, L.; Liu, Y.Y.; Zhu, B.; Li, Y.; Hua, H.; Wang, Y.H.; Zhang, J.; Jiang, Z.; Wang, Z.R. [Sichuan University, Chengdu (China). West China Medical Center. Health Ministry Key Lab. of Chronobiology], e-mail: wangzhengrong@126.com

    2009-10-15

    Period2 is a core circadian gene, which not only maintains the circadian rhythm of cells but also regulates some organic functions. We investigated the effects of mPeriod2 (mPer2) expression on radiosensitivity in normal mouse cells exposed to {sup 60}Co-{gamma}-rays. NIH 3T3 cells were treated with 12-O-tetradecanoyl phorbol-13-acetate (TPA) to induce endogenous mPer2 expression or transfected with pcDNA3.1(+)-mPer2 and irradiated with {sup 6}0Co-{gamma}-rays, and then analyzed by several methods such as flow cytometry, colony formation assay, RT-PCR, and immunohistochemistry. Flow cytometry and colony formation assay revealed that irradiated NIH 3T3 cells expressing high levels of mPer2 showed a lower death rate (TPA: 24 h 4.3% vs 12 h 6.8% and control 9.4%; transfection: pcDNA3.1-mPer2 3.7% vs pcDNA3.1 11.3% and control 8.2%), more proliferation and clonogenic survival (TPA: 121.7 {+-} 6.51 vs 66.0 {+-} 3.51 and 67.7 {+-} 7.37; transfection: 121.7 {+-} 6.50 vs 65.3 {+-} 3.51 and 69.0 {+-} 4.58) both when treated with TPA and transfected with mPer2. RT-PCR analysis showed an increased expression of bax, bcl-2, p53, cmyc, mre11, and nbs1, and an increased proportionality of bcl-2/bax in the irradiated cells at peak mPer2 expression compared with cells at trough mPer2 expression and control cells. However, no significant difference in rad50 expression was observed among the three groups of cells. Immunohistochemistry also showed increased protein levels of P53, BAX and proliferating cell nuclear antigen in irradiated cells with peak mPer2 levels. Thus, high expression of the circadian gene mPer2 may reduce the radiosensitivity of NIH 3T3 cells. For this effect, mPer2 may directly or indirectly regulate the expressions of cell proliferation- and apoptosis-related genes and DNA repair-related genes. (author)

  20. Electric-field-controlled interface strain coupling and non-volatile resistance switching of La1-xBaxMnO3 thin films epitaxially grown on relaxor-based ferroelectric single crystals

    Science.gov (United States)

    Zheng, Ming; Zhu, Qiu-Xiang; Li, Xue-Yan; Yang, Ming-Min; Wang, Yu; Li, Xiao-Min; Shi, Xun; Luo, Hao-Su; Zheng, Ren-Kui

    2014-09-01

    We have fabricated magnetoelectric heterostructures by growing ferromagnetic La1-xBaxMnO3 (x = 0.2, 0.4) thin films on (001)-, (110)-, and (111)-oriented 0.31Pb(In1/2Nb1/2)O3-0.35Pb(Mg1/3Nb1/2)O3-0.34PbTiO3 (PINT) ferroelectric single-crystal substrates. Upon poling along the [001], [110], or [111] crystal direction, the electric-field-induced non-180° domain switching gives rise to a decrease in the resistance and an enhancement of the metal-to-insulator transition temperature TC of the films. By taking advantage of the 180° ferroelectric domain switching, we identify that such changes in the resistance and TC are caused by domain switching-induced strain but not domain switching-induced accumulation or depletion of charge carriers at the interface. Further, we found that the domain switching-induced strain effects can be efficiently controlled by a magnetic field, mediated by the electronic phase separation. Moreover, we determined the evolution of the strength of the electronic phase separation against temperature and magnetic field by recording the strain-tunability of the resistance [(ΔR/R)strain] under magnetic fields. Additionally, opposing effects of domain switching-induced strain on ferromagnetism above and below 197 K for the La0.8Ba0.2MnO3 film and 150 K for the La0.6Ba0.4MnO3 film, respectively, were observed and explained by the magnetoelastic effect through adjusting the magnetic anisotropy. Finally, using the reversible ferroelastic domain switching of the PINT, we realized non-volatile resistance switching of the films at room temperature, implying potential applications of the magnetoelectric heterostructure in non-volatile memory devices.

  1. Identification of human ferritin, heavy polypeptide 1 (FTH1) and yeast RGI1 (YER067W) as pro-survival sequences that counteract the effects of Bax and copper in Saccharomyces cerevisiae.

    Science.gov (United States)

    Eid, Rawan; Boucher, Eric; Gharib, Nada; Khoury, Chamel; Arab, Nagla T T; Murray, Alistair; Young, Paul G; Mandato, Craig A; Greenwood, Michael T

    2016-03-01

    Ferritin is a sub-family of iron binding proteins that form multi-subunit nanotype iron storage structures and prevent oxidative stress induced apoptosis. Here we describe the identification and characterization of human ferritin, heavy polypeptide 1 (FTH1) as a suppressor of the pro-apoptotic murine Bax sequence in yeast. In addition we demonstrate that FTH1 is a general pro-survival sequence since it also prevents the cell death inducing effects of copper when heterologously expressed in yeast. Although ferritins are phylogenetically widely distributed and are present in most species of Bacteria, Archaea and Eukarya, ferritin is conspicuously absent in most fungal species including Saccharomyces cerevisiae. An in silico analysis of the yeast proteome lead to the identification of the 161 residue RGI1 (YER067W) encoded protein as a candidate for being a yeast ferritin. In addition to sharing 20% sequence identity with the 183 residue FTH1, RGI1 also has similar pro-survival properties as ferritin when overexpressed in yeast. Analysis of recombinant protein by SDS-PAGE and by electron microscopy revealed the expected formation of higher-order structures for FTH1 that was not observed with Rgi1p. Further analysis revealed that cells overexpressing RGI1 do not show increased resistance to iron toxicity and do not have enhanced capacity to store iron. In contrast, cells lacking RGI1 were found to be hypersensitive to the toxic effects of iron. Overall, our results suggest that Rgi1p is a novel pro-survival protein whose function is not related to ferritin but nevertheless it may have a role in regulating yeast sensitivity to iron stress.

  2. All-trans retinoic acid inhibits KIT activity and induces apoptosis in gastrointestinal stromal tumor GIST-T1 cell line by affecting on the expression of survivin and Bax protein

    Directory of Open Access Journals (Sweden)

    Taguchi Takahiro

    2010-12-01

    Full Text Available Abstract Background Imatinib, a selective tyrosine kinase inhibitor, has been used as a standard first-line therapy for irresectable and metastasized gastrointestinal stromal tumor (GIST patients. Unfortunately, most patients responding to imatinib will eventually exhibit imatinib-resistance, the cause of which is not fully understood. The serious clinical problem of imatinib-resistance demands alternative therapeutic strategy. This study was conducted to investigate the effect of all-trans retinoic acid (ATRA on GIST cell lines. Methods Cell proliferation was determined by trypan blue dye exclusion test. Western blot analysis was performed to test the expression of activated KIT, its downstream proteins, and apoptosis associated proteins. The cytotoxic interactions of imatinib with ATRA were evaluated using the isobologram of Steel and Peckham. Results and conclusion In this work, for the first time we have demonstrated that ATRA affected on cell proliferation of GIST-T1 and GIST-882 cell line through inhibition of cell growth in a dose dependent manner and induced apoptosis. High dose of ATRA induced morphologic change in GIST-T1 cells, rounded-up cells, and activated the caspase-3 protein. In further examination, we found that the ATRA-induced apoptosis in GIST-T1 cells was accompanied by the down-regulated expression of survivin and up-regulated expression of Bax protein. Moreover, ATRA suppressed the activity of KIT protein in GIST-T1 cells and its downstream signal, AKT activity, but not MAPK activity. We also have demonstrated that combination of ATRA with imatinib showed additive effect by isobologram, suggesting that the combination of ATRA and imatinib may be a novel potential therapeutic option for GIST treatment. Furthermore, the scracht assay result suggested that ATRA was a potential reagent to prevent the invasion or metastasis of GIST cells.

  3. The correlation research on the expression of Bcl-2,Bax and eNOS in the ICR mice testicles%Bcl-2和Bax在ICR小鼠睾丸中的表达及与eNOS的关联性研究

    Institute of Scientific and Technical Information of China (English)

    左俐俊; 任亚萍; 赵玮; 宋婉玲

    2016-01-01

    目的 探讨B淋巴细胞瘤/白血病-2 ( Bcl-2 )和Bcl-2相关X蛋白( Bax)在雄性ICR小鼠睾丸中的表达及与内皮型一氧化氮合酶( eNOS)的联系和意义. 方法 30只(分别为4、8、12周龄,各10只)健康雄性ICR小鼠,分为性成熟前(4周龄组)、性成熟(8周龄组)、性成熟后(12周龄组),取左侧睾丸经石蜡切片,免疫组化法检测小鼠睾丸中 eNOS、Bcl-2和Bax蛋白的表达分布情况;取右侧睾丸,Western blot法检测eNOS、Bcl-2 和 Bax的表达情况. 结果 Bcl-2 在睾丸间质细胞高表达,Bax在生精上皮有表达;8周龄小鼠睾丸间质细胞Bcl-2表达明显高于4、12周龄组,且8周龄组小鼠Bax表达明显低于4、12周龄组小鼠( P<0. 05 );4周龄组小鼠睾丸eNOS蛋白表达明显高于8、12 周龄组( P <0. 01 ).结论 Bcl-2、Bax与eNOS在睾丸间质细胞的表达并没有直接的相关性,提示NO或许未直接参与睾丸间质细胞的凋亡活动.%Objective To explore the expression and significance of the B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein ( Bax ) in ICR mice testicles, and the correlation with endothelial nitric oxide synthase (eNOS). Methods 30 (4 weeks,8 weeks and 12 weeks respectively,each 10) healthy male ICR mice were divid-ed into three groups randomly:young period,adolescent period and the period of sexual maturity. Paraffin section of the left testis was made, the expressions of the Bcl-2,Bax and eNOS in the testis of male mice were observed with immunohistochemical method. Then Western blot was carried out to screen the protein of Bcl-2,Bax and eNOS in the right side of the mice testicles. Results The Bcl-2 highly appeared in leydig cells,while Bax in rawhide cell. The expression of Bcl-2 in the 8-week-old mice leydig cells was significantly higher than that in 4 or 12-week-old groups. The protein levels of Bax in the 8-week-old mice was lower than that in 4 or 12-week-old group ( P <0. 05). Besides,the expression of eNOS in 4-week

  4. Effect of thermal stress on expression profile of apoptosis related genes in peripheral blood mononuclear cells of transition Sahiwal cow.

    Science.gov (United States)

    Somal, A; Aggarwal, A; Upadhyay, R C

    2015-01-01

    The study was conducted to evaluate the effect of thermal stress on expression profile of genes related to apoptosis in peripartum Sahiwal cows. For this, twelve pregnant dry Sahiwal cows were selected from Livestock Research Centre at National Dairy Research Institute, Karnal. The cows were divided into two groups consisting of six Sahiwal cows each. Cows of group I calved during thermoneutral temperature conditions (THI=67.3) and cows of group II calved in summer season (THI=79.9). Blood samples were collected on -15, 0 and +15 days with respect to calving where day '0' represents the day of calving. The peripheral blood mononuclear cells (PBMC) were separated and total RNA was isolated for the BCL-2 (B-Cell Lymphoma-2), BAX (BCL-2 antagonist killer-1), BAK (Bcl-2-associated X protein), CASP-3 (cysteine-aspartic proteases-3) and P53 (tumour protien-53) mRNAs expression. It was found that there was up regulation of CASP-3 on the day of calving during both temperature conditions. Comparison between the two temperature conditions showed that expression of CASP-3, BCL-2, BAK, P53 and ratio of BAX/BCL-2 in PBMC increased during summer as compared to thermoneutral condition suggesting the susceptibility of these cells to apoptosis. Based on the above findings it can be concluded that during calving PBMC are more susceptible to apoptosis, and summer being more stressful potentiates the apoptosis of PBMC in Sahiwal cows.

  5. A gold nanoparticle pentapeptide: gene fusion to induce therapeutic gene expression in mesenchymal stem cells.

    Science.gov (United States)

    Muroski, Megan E; Morgan, Thomas J; Levenson, Cathy W; Strouse, Geoffrey F

    2014-10-22

    Mesenchymal stem cells (MSC) have been identified as having great potential as autologous cell therapeutics to treat traumatic brain injury and spinal injury as well as neuronal and cardiac ischemic events. All future clinical applications of MSC cell therapies must allow the MSC to be harvested, transfected, and induced to express a desired protein or selection of proteins to have medical benefit. For the full potential of MSC cell therapy to be realized, it is desirable to systematically alter the protein expression of therapeutically beneficial biomolecules in harvested MSC cells with high fidelity in a single transfection event. We have developed a delivery platform on the basis of the use of a solid gold nanoparticle that has been surface modified to produce a fusion containing a zwitterionic, pentapeptide designed from Bax inhibiting peptide (Ku70) to enhance cellular uptake and a linearized expression vector to induce enhanced expression of brain-derived neurotrophic factor (BDNF) in rat-derived MSCs. Ku70 is observed to effect >80% transfection following a single treatment of femur bone marrow isolated rat MSCs with efficiencies for the delivery of a 6.6 kbp gene on either a Au nanoparticle (NP) or CdSe/ZnS quantum dot (QD). Gene expression is observed within 4 d by optical measurements, and secretion is observed within 10 d by Western Blot analysis. The combination of being able to selectively engineer the NP, to colocalize biological agents, and to enhance the stability of those agents has provided the strong impetus to utilize this novel class of materials to engineer primary MSCs. PMID:25198921

  6. Bioassay-guided fractionation of extracts from Codiaeum variegatum against Entamoeba histolytica discovers compounds that modify expression of ceramide biosynthesis related genes.

    Science.gov (United States)

    Mfotie Njoya, Emmanuel; Weber, Christian; Hernandez-Cuevas, Nora Adriana; Hon, Chung-Chau; Janin, Yves; Kamini, Melanie F G; Moundipa, Paul F; Guillén, Nancy

    2014-01-01

    Leaves of Codiaeum variegatum ("garden croton") are used against bloody diarrhoea by local populations in Cameroon. This study aims to search for the active components from C. variegatum against Entamoeba histolytica, and thereby initiate the study of their mechanism of action. A bioassay-guided screening of the aqueous extracts from C. variegatum leaves and various fractions was carried out against trophozoites of E. histolytica axenic culture. We found that the anti-amoebic activity of extracts changed with respect to the collection criteria of leaves. Thereby, optimal conditions were defined for leaves' collection to maximise the anti-amoebic activity of the extracts. A fractionation process was performed, and we identified several sub-fractions (or isolated compounds) with significantly higher anti-amoebic activity compared to the unfractionated aqueous extract. Anti-amoebic activity of the most potent fraction was confirmed with the morphological characteristics of induced death in trophozoites, including cell rounding and lysis. Differential gene expression analysis using high-throughput RNA sequencing implies the potential mechanism of its anti-amoebic activity by targeting ceramide, a bioactive lipid involved in disturbance of biochemical processes within the cell membrane including differentiation, proliferation, cell growth arrest and apoptosis. Regulation of ceramide biosynthesis pathway as a target for anti-amoebic compounds is a novel finding which could be an alternative for drug development against E. histolytica. PMID:24416462

  7. Bioassay-guided fractionation of extracts from Codiaeum variegatum against Entamoeba histolytica discovers compounds that modify expression of ceramide biosynthesis related genes.

    Directory of Open Access Journals (Sweden)

    Emmanuel Mfotie Njoya

    Full Text Available Leaves of Codiaeum variegatum ("garden croton" are used against bloody diarrhoea by local populations in Cameroon. This study aims to search for the active components from C. variegatum against Entamoeba histolytica, and thereby initiate the study of their mechanism of action. A bioassay-guided screening of the aqueous extracts from C. variegatum leaves and various fractions was carried out against trophozoites of E. histolytica axenic culture. We found that the anti-amoebic activity of extracts changed with respect to the collection criteria of leaves. Thereby, optimal conditions were defined for leaves' collection to maximise the anti-amoebic activity of the extracts. A fractionation process was performed, and we identified several sub-fractions (or isolated compounds with significantly higher anti-amoebic activity compared to the unfractionated aqueous extract. Anti-amoebic activity of the most potent fraction was confirmed with the morphological characteristics of induced death in trophozoites, including cell rounding and lysis. Differential gene expression analysis using high-throughput RNA sequencing implies the potential mechanism of its anti-amoebic activity by targeting ceramide, a bioactive lipid involved in disturbance of biochemical processes within the cell membrane including differentiation, proliferation, cell growth arrest and apoptosis. Regulation of ceramide biosynthesis pathway as a target for anti-amoebic compounds is a novel finding which could be an alternative for drug development against E. histolytica.

  8. The effects of the Reinforcing Kidney and Activating Blood Recipe Affecting the Value of bcl-2, bax and caspase-3 in the Tissue of Corpus Cavernosum Smooth Muscle of DMED- Rats%补肾活血合剂对糖尿病阳痿大鼠阴茎平滑肌组织中Bcl-2 Bax和Caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    张国豪; 方再军; 黄青松

    2011-01-01

    目的:观察补肾活血合剂对糖尿病阳痿(DMED)大鼠阴茎平滑肌组织中Bcl-2、Bax、Caspase-3表达的影响.方法:将实验小鼠用2%链脲佐菌素液(STZ)接60mg/kg,腹腔注射建立糖尿病模型,然后在每只糖尿病模型大鼠颈项处注射阿朴吗啡(APOsigma公司)80μg/kg,录相记录阴茎勃起次数.筛选DMED模型,将造成DMED模型随机分为正常组、模型组、中药高、低剂量组、达美康组、达美康+安雄组.除正常组外,其余各组连续给药12周,然后处死测定阴茎平滑肌组织中Bcl-2,Bax,Caspase-3的表达情况.结果:补肾活血合剂对Bc1-2,Bax,Caspase-3有改善作用.结论:补肾活血合剂对糖尿病阳痿大鼠阴茎平滑肌组织中Bcl-2,Bax,Caspase-3有调控效果.%Objective: Observing the effects of reinforcing kidney and activating blood recipe affecting the value of bcl - 2, bax and caspase - 3 in the tissue of corpus cavernosum Smooth muscle of DMED - rats. Methods: An animal model of diabetes was induced by a single intravenous dose of streptozotocin (STZ, 60mg/kg body weight) in Wistar rats. Recording the numbers of penile erection of each rat by camera after subcutaneous inject of apomorphine (80mg/kg).Screening the model of STZ - rats with erectile dysfunction and grouping into 6 groups: control group, model group, Chinese Medicine group, low- dosage group, Diamicron group, Diamicron combined Testosterone Undecanoate group. After administrating 12 weeks, observed the value of bcl -2, bax and caspase-3 in the tissue of corpus cavernosum smooth muscle of DMED - rats. Results: The reinforcing kidney and activating blood recipe can improve the vulue of bcl - 2,bax and caspase -3. Conclusion: The reinforcing kidney and activating blood recipe has regulatory function for bcl -2,bax and caspase - 3 in the tissue of corpus cavernosum smooth muscle of DMED - rats.

  9. Genes of cell-cell interactions, chemotherapy detoxification and apoptosis are induced during chemotherapy of acute myeloid leukemia

    International Nuclear Information System (INIS)

    The molecular changes in vivo in acute myeloid leukemia cells early after start of conventional genotoxic chemotherapy are incompletely understood, and it is not known if early molecular modulations reflect clinical response. The gene expression was examined by whole genome 44 k oligo microarrays and 12 k cDNA microarrays in peripheral blood leukocytes collected from seven leukemia patients before treatment, 2–4 h and 18–24 h after start of chemotherapy and validated by real-time quantitative PCR. Statistically significantly upregulated genes were classified using gene ontology (GO) terms. Parallel samples were examined by flow cytometry for apoptosis by annexin V-binding and the expression of selected proteins were confirmed by immunoblotting. Significant differential modulation of 151 genes were found at 4 h after start of induction therapy with cytarabine and anthracycline, including significant overexpression of 31 genes associated with p53 regulation. Within 4 h of chemotherapy the BCL2/BAX and BCL2/PUMA ratio were attenuated in proapoptotic direction. FLT3 mutations indicated that non-responders (5/7 patients, 8 versus 49 months survival) are characterized by a unique gene response profile before and at 4 h. At 18–24 h after chemotherapy, the gene expression of p53 target genes was attenuated, while genes involved in chemoresistance, cytarabine detoxification, chemokine networks and T cell receptor were prominent. No signs of apoptosis were observed in the collected cells, suggesting the treated patients as a physiological source of pre-apoptotic cells. Pre-apoptotic gene expression can be monitored within hours after start of chemotherapy in patients with acute myeloid leukemia, and may be useful in future determination of therapy responders. The low number of patients and the heterogeneity of acute myeloid leukemia limited the identification of gene expression predictive of therapy response. Therapy-induced gene expression reflects the complex

  10. Microarray profiling for differential gene expression in PMSG-hCG stimulated preovulatory ovarian follicles of Chinese Taihu and Large White sows

    Directory of Open Access Journals (Sweden)

    Xiong Yuanzhu

    2011-02-01

    Full Text Available Abstract Background The Chinese Taihu is one of the most prolific pig breeds in the world, which farrows at least five more piglets per litter than Western pig breeds partly due to a greater ovulation rate. Variation of ovulation rate maybe associated with the differences in the transcriptome of Chinese Taihu and Large White ovaries. In order to understand the molecular basis of the greater ovulation rate of Chinese Taihu sows, expression profiling experiments were conducted to identify differentially expressed genes in ovarian follicles at the preovulatory stage of a PMSG-hCG stimulated estrous cycle from 3 Chinese Taihu and 3 Large White cycling sows by using the Affymetrix Porcine Genechip™. Results One hundred and thirty-three differentially expressed genes were identified between Chinese Taihu and Large White sows by using Affymetrix porcine GeneChip (p ≤ 0.05, Fold change ≥ 2 or ≤ 0.5. Gene Ontology (GO analysis revealed that these genes belonged to the class of genes that participated in regulation of cellular process, regulation of biological process, biological regulation, developmental process, cell communication and signal transduction and so on. Significant differential expression of 6 genes including WNT10B and DKK2 in the WNT signaling pathway was detected. Real-time RT-PCR confirmed the expression pattern in seven of eight selected genes. A search of chromosomal location revealed that 92 differentially expressed transcripts located to the intervals of quantitative trait loci (QTLs for reproduction traits. Furthermore, SNPs of two differentially expressed genes- BAX and BMPR1B were showed to be associated with litter size traits in Large White pigs and Chinese DIV line pigs (p ≤ 0.1 or p ≤ 0.05. Conclusions Our study detected many genes that showed differential expression between ovary follicles of two divergent breeds of pigs. Genes involved with regulation of cellular process, regulation of biological process, in

  11. Visualizing spatiotemporal dynamics of apoptosis after G1 arrest by human T cell leukemia virus type 1 Tax and insights into gene expression changes using microarray-based gene expression analysis

    Directory of Open Access Journals (Sweden)

    Arainga Mariluz

    2012-06-01

    Full Text Available Abstract Background Human T cell leukemia virus type 1 (HTLV-1 Tax is a potent activator of viral and cellular gene expression that interacts with a number of cellular proteins. Many reports show that Tax is capable of regulating cell cycle progression and apoptosis both positively and negatively. However, it still remains to understand why the Tax oncoprotein induces cell cycle arrest and apoptosis, or whether Tax-induced apoptosis is dependent upon its ability to induce G1 arrest. The present study used time-lapse imaging to explore the spatiotemporal patterns of cell cycle dynamics in Tax-expressing HeLa cells containing the fluorescent ubiquitination-based cell cycle indicator, Fucci2. A large-scale host cell gene profiling approach was also used to identify the genes involved in Tax-mediated cell signaling events related to cellular proliferation and apoptosis. Results Tax-expressing apoptotic cells showed a rounded morphology and detached from the culture dish after cell cycle arrest at the G1 phase. Thus, it appears that Tax induces apoptosis through pathways identical to those involved in G1 arrest. To elucidate the mechanism(s by which Tax induces cell cycle arrest and apoptosis, regulation of host cellular genes by Tax was analyzed using a microarray containing approximately 18,400 human mRNA transcripts. Seventeen genes related to cell cycle regulation were identified as being up or downregulated > 2.0-fold in Tax-expressing cells. Several genes, including SMAD3, JUN, GADD45B, DUSP1 and IL8, were involved in cellular proliferation, responses to cellular stress and DNA damage, or inflammation and immune responses. Additionally, 23 pro- and anti-apoptotic genes were deregulated by Tax, including TNFAIP3, TNFRS9, BIRC3 and IL6. Furthermore, the kinetics of IL8, SMAD3, CDKN1A, GADD45A, GADD45B and IL6 expression were altered following the induction of Tax, and correlated closely with the morphological changes observed by time

  12. Sundew plant, a potential source of anti-inflammatory agents, selectively induces G2/M arrest and apoptosis in MCF-7 cells through upregulation of p53 and Bax/Bcl-2 ratio.

    Science.gov (United States)

    Ghate, N B; Das, A; Chaudhuri, D; Panja, S; Mandal, N

    2016-01-01

    The worldwide cancer incidences are remarkable despite the advancement in cancer drug discovery field, highlighting the need for new therapies focusing on cancer cell and its microenvironment, including inflammation. Several species of Drosera (family: Droseraceae) are used in various traditional as well as homeopathic systems of medicine. Drosera burmannii Vahl. is also enlisted in French Pharmacopoeia in 1965 for the treatment of inflammatory diseases, including chronic bronchitis, asthma and whooping cough. The present study is designed to substantiate the potential of D. burmannii in in vitro anticancer activity and its relation with anti-inflammatory property. In vitro anticancer study revealed that DBME is inhibiting the proliferation of MCF-7 cells without affecting the viability of other malignant and non-malignant cells. DBME induced G2/M phase arrest and apoptosis in MCF-7 cells by suppressing the expression of cyclin A1, cyclin B1 and Cdk-1 and increasing the expression of p53, Bax/Bcl-2 ratio leading to activation of caspases and PARP degradation. Presence of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors alone did prevent the apoptosis partially while apoptosis prevention was significantly observed when used in combination, suggesting vital role of caspases in DBME-induced apoptosis in MCF-7 cells. DBME also downregulated LPS-induced increased expression of iNOS, COX-2 and TNF-α along with suppression on intracellular ROS production that confirms the potential of DBME as anti-inflammatory extract. GCMS analysis revealed the presence of four major compounds hexadecanoic acid, tetradecanoic acid, hexadecen-1-ol, trans-9 and 1-tetradecanol along with some other fatty acid derivatives and carotenoids (Beta-doradecin) in DBME. These findings confirmed the anti-inflammatory activity of DBME, which is already listed in French Pharmacopeia in 1965. Here we have additionally reported the anti-breast cancer activity of DBME and its relation to the

  13. The expression of Bax, Bcl-2 and NF-κB in the early stage of liver regeneration%Bax、Bcl-2和NF-κB在肝再生早期中的表达

    Institute of Scientific and Technical Information of China (English)

    杜赵康; 杨开明

    2014-01-01

    目的 研究在大鼠大部肝切除(partial hepatectomy,PH)术后细胞凋亡调节基因(Bcl-2 associated X protein Bax)、Bcl-2(B-cell lymphoma-2)及NF-κB(nuclear factor-kappa B)三者的分布和表达,探讨三者在肝再生早期中的调节机制及其相互调控作用.方法 采用SD大鼠35只分7组,每组5只构建大鼠肝脏再生模型,并在显微镜下观察肝大部切除后早期(0.5、1、4、6、8、12、24 h)肝组织的形态学变化,采用免疫组织化学SABC法检测Bax、Bcl-2、NF-κB在正常肝组织中的表达,并研究在肝再生早期中的分布及表达变化.结果 Bax、Bcl-2、NF-κB在正常肝组织未见表达,但在PH后30 min,Bax、Bcl-2及NF-κB即在肝细胞和胆管上皮细胞和肝血窦内皮内开始出现表达,PH后6h表达达到高峰,之后其表达逐渐下降,而Bcl-2的表达一直保持在较高水平.NF-κB于PH后6h表达出现高峰后其表达逐渐下调,24h时NF-κB表达上调,出现另一表达高峰.结论 肝大部切除后再生早期,存在着凋亡和抑制凋亡的分子调控机制,NF-κB的表达可能与激活Bcl-2、抑制肝细胞的凋亡从而促进肝细胞再生有关.

  14. Sundew plant, a potential source of anti-inflammatory agents, selectively induces G2/M arrest and apoptosis in MCF-7 cells through upregulation of p53 and Bax/Bcl-2 ratio

    Science.gov (United States)

    Ghate, NB; Das, A; Chaudhuri, D; Panja, S; Mandal, N

    2016-01-01

    The worldwide cancer incidences are remarkable despite the advancement in cancer drug discovery field, highlighting the need for new therapies focusing on cancer cell and its microenvironment, including inflammation. Several species of Drosera (family: Droseraceae) are used in various traditional as well as homeopathic systems of medicine. Drosera burmannii Vahl. is also enlisted in French Pharmacopoeia in 1965 for the treatment of inflammatory diseases, including chronic bronchitis, asthma and whooping cough. The present study is designed to substantiate the potential of D. burmannii in in vitro anticancer activity and its relation with anti-inflammatory property. In vitro anticancer study revealed that DBME is inhibiting the proliferation of MCF-7 cells without affecting the viability of other malignant and non-malignant cells. DBME induced G2/M phase arrest and apoptosis in MCF-7 cells by suppressing the expression of cyclin A1, cyclin B1 and Cdk-1 and increasing the expression of p53, Bax/Bcl-2 ratio leading to activation of caspases and PARP degradation. Presence of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors alone did prevent the apoptosis partially while apoptosis prevention was significantly observed when used in combination, suggesting vital role of caspases in DBME-induced apoptosis in MCF-7 cells. DBME also downregulated LPS-induced increased expression of iNOS, COX-2 and TNF-α along with suppression on intracellular ROS production that confirms the potential of DBME as anti-inflammatory extract. GCMS analysis revealed the presence of four major compounds hexadecanoic acid, tetradecanoic acid, hexadecen-1-ol, trans-9 and 1-tetradecanol along with some other fatty acid derivatives and carotenoids (Beta-doradecin) in DBME. These findings confirmed the anti-inflammatory activity of DBME, which is already listed in French Pharmacopeia in 1965. Here we have additionally reported the anti-breast cancer activity of DBME and its relation to the

  15. Circadian transitions in radiation dose-dependent augmentation of mRNA levels for DNA damage-induced genes elicited by accurate real-time RT-PCR quantification

    International Nuclear Information System (INIS)

    Molecular mechanisms of intracellular response after DNA-damage by exposure to ionizing radiation have been studied. In the case of cells isolated from living body of human and experimental animals, alteration of the responsiveness by physiological oscillation such as circadian rhythm must be considered. To examine the circadian variation in the response of p53-responsible genes p21, mdm2, bax, and puma, we established a method to quantitate their mRNA levels with high reproducibility and accuracy based on real-time reverse transcription polymerase chain reaction (RT-PCR) and compared the levels of responsiveness in mouse hemocytes after diurnal irradiation to that after nocturnal irradiation. Augmentations of p21 and mdm2 mRNA levels with growth-arrest and of puma mRNA before apoptosis were confirmed by time-course experiment in RAW264.7, and dose-dependent increases in the peak levels of all the RNA were shown. Similarly, the relative RNA levels of p21, mdm2, bax, and puma per glyceraldehyde-3-phosphate dehydrogenase (GAPDH) also increased dose-dependently in peripheral blood and bone marrow cells isolated from whole-body-irradiated mice. Induction levels of all messages reduced by half after nighttime irradiation as compared with daytime irradiation in blood cells. In marrow cells, nighttime irradiation enhanced the p21 and mdm2 mRNA levels than daytime irradiation. No significant difference in bax or puma mRNA levels was observed between nighttime and daytime irradiation in marrow cells. This suggests that early-stage cellular responsiveness in DNA damage-induced genes is modulated between diurnal and nocturnal irradiation. (author)

  16. Patterns of expression of cell cycle/apoptosis genes along the spectrum of thyroid carcinoma progression

    NARCIS (Netherlands)

    B. Saltman; B. Singh; C.V. Hedvat; V.B. Wreesmann; R. Ghossein

    2006-01-01

    Background. Genetic screening studies suggest that genetic changes underlie progression from well differentiated, to anoplastic thyroid cancers. The aim of this study is to determine to what extent cell cycle/apoptosis regulators contribute to cancer progression. Methods. Tissue microarrarys (TMAs)

  17. Bax、Bcl-2在局部晚期宫颈癌中的表达及意义%Studies on the Expression and Clinical Significance of Bax and Bcl-2 before neoadjuvant chemotherapy in Local y advanced cervical cancer

    Institute of Scientific and Technical Information of China (English)

    夏红; 易建华; 杨丽; 褚桂芬

    2014-01-01

    Objective To investigate the expression and clinical significance of Bax and Bcl-2 before neo-adjuvant chemotherapy (NACT) in local y advanced cervical cancer (LACC) . Methods 47 patients with stage ⅠB2-ⅡB of LACC treated with neoadjuvant chemotherapy(NACT) between January 2010 and December 2013 in our hospital were retrospectively analyzed. The expressions of Bax and Bcl-2 were determined by means of immunohistochemistry before NACT. Results: The total effective rate of NACT was 72.3%, including CR 14.9%. The Bax and Bcl-2 expression was 55.3% and 36.2% before NACT. In patients who react and react poorly to NACT, the expression of Bax was 64.7% and 38.5% (p0.05). The staining intensity for Bax was related to histopathological grade and lymphatic metastasis (p0.05). The staining intensity for Bcl-2 was not related to the above clinical and pathological factor (P>0.05). Conclusions The NACT of paclitaxel and cisplatin was effective to LACC. The expression of Bax could be related with biological behavior of the tumor and considered as the index to predict efficacy in NACT.%目的:探讨Bax、Bcl-2蛋白在局部晚期宫颈癌(LACC)中的表达及其临床意义。方法应用免疫组化检测47例LACC 组织中化疗前Bax、Bcl-2蛋白的表达情况。结果①47例LACC中,NACT有效率为72.3%,CR 为14.9%。②化疗前,Bax及Bcl-2蛋白阳性表达率分别为:55.3%、36.2%。临床有效及无效病例中,B a x阳性表达率分别为64.7%、38.5%,差异有统计学意义(P0.05)。③肿瘤低分化患者中B a x阳性表达率明显低于高中分化患者,有淋巴结转移者明显低于无淋巴结转移者(P0.05)。Bcl-2表达在各临床病理参数方面均无明显差异(P>0.05)。结论紫杉醇+顺铂静脉化疗对LACC有效,Bax蛋白异常表达与宫颈癌的生物学行为有关,可作为预测和判定NACT疗效的指标。

  18. Investigation of benazepril protection on diabetic myocardium by downregulating the expression of TGF-β1 and Bax%贝那普利下调TGF-β1及Bax表达对糖尿病大鼠心肌的保护作用

    Institute of Scientific and Technical Information of China (English)

    符丽娟; 杨育红; 包翠芬; 刘婉珠

    2011-01-01

    Objective To investigate the effect of benazepril (BZ) on the expression of TGF-β1 and Bax on diabetic myocardium and myocardial hypertrophy. Methods Diabetic animal models were induced by injecting streptozotocin. Model rats were divided into diabetes and BZ groups, control group was also established. BZ was given to treatment group at dose of 10 mg/kg each day by gavage. At 8 weeks, heart mass indexes (HMI) and left ventricular myocardium mass indexes (LVMI) were measured. The expressions of TGF-β1 and Bax in cardiomyocyte were detected and myocardium was studied by electronic microscope. Results Compared with control group, HMI and LVMI were significantly increased in diabetes group ( P < 0. 05 ). The expression of TGF-β1 and Bax were strengthened in diabetic myocardium. Electron microscopic morphometry of heart samples revealed typical diabetic alterations consisting in a focal loss, disorganization of myofihril, mitochondria swelling, collagen proliferation in extracellular matrix. After treatment with BZ for 8 weeks, HMI and LVMI were decreased ( P < 0. 01 ). The expression of TGF-β1 and Bax were weakened and myocardial ultrastructure was better than that of diabetes group. Conclusions TGF-β1 can induce myocardial apoptosis by stimulating the expression of Bax and promote diabetic cardiac hypertrophy, BZ can improve the cardiac function partly due to its effects of downregulating the expression of TGF-β1 and Bax.%目的 探讨贝那普利(BZ)对糖尿病(DM)大鼠心肌转化生长因子β1(TGF-β1)和Bax的表达及心肌肥厚指标的影响.方法 SD大鼠腹腔注射链脲佐菌素(STZ)制备DM模型.实验分为正常对照组(CON)、DM组和BZ治疗组(BZ).治疗组每日灌胃给予贝那普利(10 mg·kg-1·d-1).8 w后测定心脏质量指数(HMI)、左心室质量指数(LVMI),并取左心室心肌免疫组化法检测TGF-β1和 Bax在心肌组织中的表达,透射电镜观察心肌组织超微结构变化.结果 与CON组相比,DM组大

  19. C57/BL6小鼠海马发育和老化过程中Bcl-2及Bax的表达%Expression of Bcl-2 and Bax protein in the development and aging of C57/BL6 mouse hippocampus

    Institute of Scientific and Technical Information of China (English)

    李潮; 张敬坤; 张莉

    2011-01-01

    目的 观察C57/BL6小鼠海马发育老化过程中Bcl-2、Bax蛋白的表达变化.方法 取不同胚龄(embryonic day,E)、生后(postnatal day,P)日龄或月龄的小鼠海马,应用免疫组化及图像分析技术检测Bcl 2和Bax蛋白的表达.结果 E16 d~P7 d,海马CA区锥体细胞层和齿状回颗粒细胞层Bcl 2和Bax阳性细胞均逐渐增多,平均吸光度( average optical,AO)值逐渐增大;P14 d~P21 d阳性细胞逐渐减少,CA区AO值逐渐减小,齿状回AO值先增大后减小;P28 d后趋于稳定.CA区Bcl 2/Bax比值E16 d~E20 d增大,P1 d减小,P3 d~P5 d增大,P7 d~P21 d减小;以后趋于稳定.齿状回Bcl 2/Bax比值只在P1 d降低.结论 Bcl-2和Bax蛋白可能与小鼠海马发育和老化过程中的形态学变化密切相关.%Objective To observe the changes of expression of Bcl-2 and Bax protein in the development and aging of C57/BL6 mouse hippocampus. Methods Immunohistochemical and image analysis methods were used to detect the expression of Bcl-2 and Bax protein in the mouse hippocampus of different age. Results From embyronic (E) 16 day (d) to postnatal day (P) 7 d, Bcl-2 and Bax immunoreactive cells increased in the pyramidal layer and granular layer, and the average optical (AO) increased too. From P14 d to P21 d, immunoreactive cells decreased, and their AO decreased gradually in the CA area; AO increased first but then decreased in dentate gyrus. On P28 d, the immunoreactive cells and their AO were unchanged. Moreover, the ratio of Bcl-2/Bax rose on E16 d to E20 d, diminished on P1 d, increased on P3 d to P5 d, and decreased on P7 d to P21 d. Then the ratio of Bcl-2/ Bax remained unchanged after P28 d. Conclusion Bcl-2 and Bax protein may be correlated with the morphological changes in the development and aging of mouse hippocampus.

  20. Expression of Cell Apoptosis Protein-Bax in Locally Advanced Cervical Carcinoma after Neoadjuvant Chemotherapy as an Indicator of the Efficiency of the Therapy%局部晚期宫颈癌新辅助化疗后Bax蛋白作为治疗有效性的指标

    Institute of Scientific and Technical Information of China (English)

    游艳琴; 宋磊

    2006-01-01

    探讨Bax蛋白与宫颈癌新辅助化疗疗效之间的关系,用免疫组化方法检测23例局部晚期宫颈癌组织新辅助化疗前后Bax蛋白的表达.结果化疗后2例为临床完全缓解,17例为临床部分缓解,4例患者对新辅助化疗无反应,总有效率为82.61%.NACT前有反应者和无反应者宫颈癌组织的Bax表达无统计学差异(P>0.05).对化疗有反应者NACT后Bax WS值有显著性差异(P<0.05);无反应者NACT后Bax WS值无统计学差异(P>0.05).不同病理类型和临床分期患者NACT前后Bax变化值无显著性差异(P>0.05);不同病理分级患者NACT前后Bax变化值有统计学差异(P<0.05).说明Bax与化疗疗效有显著相关性.NACT前后Bax变化值与临床分期和病理类型无关,而与病理分级有显著相关性.Bax有可能作为判断化疗疗效的指标.

  1. Genes and Psoriasis

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    ... Diet Tips" to find out more! Email * Zipcode Genes and Psoriasis Genes hold the key to understanding ... is responsible for causing psoriatic disease. How do genes work? Genes control everything from height to eye ...

  2. Genes and Hearing Loss

    Science.gov (United States)

    ... Meeting Calendar Find an ENT Doctor Near You Genes and Hearing Loss Genes and Hearing Loss Patient ... mutation may only have dystopia canthorum. How Do Genes Work? Genes are a road map for the ...

  3. Effects of Zuogui Pill and Yougui Pill on Expression of Apoptosis Protein Fas and Bax of Brain Tissue in Rat with Experimental Autoimmune Encephalomyelitis%左归丸和右归丸对自身免疫性脑脊髓炎大鼠脑组织中凋亡蛋白Fas、 Bax表达的影响

    Institute of Scientific and Technical Information of China (English)

    寇爽; 王义周; 李明; 齐放; 郑琦; 赵晖; 王蕾

    2011-01-01

    Objective:To observe the effects of Zuogui Pill and Yougui Pill on the expression of apoptosis protein Fas and Bax of brain tissue in rat with experimental autoimmune encephalomyelitis ( EAE ) .Methods: The Lewis rat were immunized with myelin basic protein ( MBP ) 68-86 to be made EAE model. The animals were measured the body weight, temperature, volume of food and drink as well as graded daily for clinical disability. The rats were observed the incidence, incubation period, mortality rate and the change of disease course. Brain and spinal cord of the rats were harvested and the pathological changes were studied after dying by HE staining. The expression of Fas and Bax in brain and spinal cord of rats were detected by the method by immunohistochemisty. Results: Zuogui Pill and Yougui Pill can relieve the infiltrated inflammatory cells around focal zone, and inhibited the expression of Fas and Bax, similarly the hormone. Conclusion: Zuogui Pill and Yougui Pill have the effects on prevent and treatment the rats with EAE, the mechanism may be related with regulating expression of apoptosis protein Fas and Bax.%目的:观察左、右归丸对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE )大鼠脑组织中凋亡蛋白Fas、Bax表达的影响.方法:用髓鞘碱性蛋白(myelin basic proteir68-86,MBP68-86)免疫Lewis 大鼠建立EAE模型.观察各组大鼠体重体温变化,饮食和饮水变化,神经功能评分,发病率、死亡率、潜伏期及病程变化.取脑和脊髓,HE染色后进行病理观察;用免疫组化法检测大鼠脑组织中Fas、Bax的表达情况.结果:左归丸组和右归丸组明显减轻病灶区域的炎性细胞浸润,对Fas、Bax的表达均有一定的抑制作用,与激素组类似.结论:左、右归丸均有防治小鼠EAE的作用,其作用机制可能与调节细胞凋亡蛋白Fas、Bax的表达有关.

  4. Inhibitory effect and affect on Bcl-2 and Bax protein expression of renal cancer prescription No.1 in mice with renal cancer%解氏肾癌一号方对小鼠肾癌的抑制作用及对Bcl-2和Bax蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    朱成功; 崔佳; 赵莹莹; 解建国

    2015-01-01

    Objective To investigate inhibitory effect and affect on Bcl-2 and Bax protein expression of renal cancer prescription No.1 in mice with renal cancer. Methods The animal models of renal cancer were established and divided into saline control group,Chinese medicine control group,interleukin-2 group and renal cancer prescription NO.1 group. Inhibitory rate of tumor in four groups was calculated and the apoptosis-related protein Bcl-2 and Bax index were de-tected by immuno- histochemistry. Results The inhibitory rate of tumor in renal cancer prescription NO.1 group was higher than that in saline control group and interleukin-2 group respectively,and the weight of mice was increased.The expression of Bcl-2 in renal cancer prescription NO.1 group was lower,but expression of Bax in renal cancer prescrip-tion NO.1 group was higher. Conclusion Renal cancer prescription NO.1 can inhibit the expression of Bcl-2,raise the expression of Bax,and suppress tumor growth,improve the quality of life in mice.%目的:探讨解氏肾癌一号方对小鼠肾癌的抑制作用及对Bcl-2和Bax蛋白表达的影响。方法建立肾癌小鼠动物模型,分为生理盐水对照组、中药对照组、白介素-2组、解氏肾癌一号方组,计算各组抑瘤率以及采用免疫组化法检测凋亡相关蛋白Bcl-2和Bax的表达。结果解氏肾癌一号方组抑瘤率高于生理盐水对照组及白介素-2组,小鼠体重增加。解氏肾癌一号方组小鼠肿瘤组织Bcl-2表达下调,Bax表达上调。结论解氏肾癌一号方可以下调Bcl-2的表达,上调Bax的表达,从而抑制肿瘤生长,改善小鼠的生存质量。

  5. PEP-1-SOD1对离体缺血再灌注损伤大鼠心肌Bax、Bcl-2蛋白表达的影响%Effects of PEP-1-SOD1 on Expression of Bax,Bcl-2 of Myocardium in Rats Induced with Ischemia-Reperfusion Injury ex vivo

    Institute of Scientific and Technical Information of China (English)

    柯尊平; 王家宁; 王磊; 唐俊明; 杨建业; 黄永章; 张宏考

    2011-01-01

    Objective To investigate the effects of PEP-1 mediated human Cu,Zn superoxide dismutase on the myocardium apoptosis induced by ischemia-reperfusion injury(IRI) in rats ex vivo.Methods The MIRI rats model were prepared with Langendorff perfusion system ex vivo.The model rats were randomly divided into control group ,SOD1 pretreated group,PEP1-SOD1 pretreated with 25,50,100 μmol/L groups.The myocardium apoptosis and expression of Bax, Bcl-2 were determined with TUNEL method and immunofluorescencein method after reperfusion.Results Compared with control and SOD1 groups,the apoptosis indexes(AI) were significantly decreased,the expression of Bcl-2 were upregulated,the expression of Bax were downregulated in all PEP-1-SOD1 groups( all P < 0.01 ).Conclusion The fusion protein PEP-1-SOD1 could inhibit the apoptosis of myocardium in MIRI rats,which may be related to its upregulation of Bax expression and downregulation of Bcl-2 expression.%目的:研究细胞穿透肽PEP-1介导人铜锌-超氧化物歧化酶(cu,Zn-SOD,SOD1)对大鼠离体心肌缺血再灌注损伤(MIRI)细胞凋亡的影响.方法:采用Langendorff灌流系统对离体大鼠心脏进行停灌-复灌建立心肌缺血再灌注损伤模型.大鼠随机分为对照组、SODI蛋白预处理组,25、50、100μmol/L PEP-1-SOD1蛋白预处理组.复灌结束后,TUNEL法检测心肌细胞凋亡,免疫荧光法检测心肌组织Bax、Bel-2蛋白表达.结果:与对照组及SOD1组相比,各PEP-1-SOD1组心肌细胞凋亡指数(AI)显著下降,Bcl-2蛋白表达升高,Bax蛋白表达显著减少(P<0.01).结论:PEP-1-SOD1融合蛋白可抑制离体心脏缺血再灌注损伤大鼠心肌细胞凋亡,其机制可能与上调Bax表达及下调Bcl-2表达有关.

  6. Der Einfluss des α2-Adrenozeptor Agonisten Dexmedetomidin auf die Expression Apoptose-assoziierter Proteine nach inkompletter zerebraler Hemisphärenischämie bei der Ratte im protrahierten zeitlichen Verlauf

    OpenAIRE

    Zintner, Susanne

    2005-01-01

    Long-term effects of Dexmedetomidin on the expression of apoptosis-regulating proteins after incomplete cerebral ischemia in rats This study investigates the effect of the α2-adrenozeptor agonist Dexmedetomidine on the expression of the apoptosis-regulating proteins Bax, p53, Bcl-2 and Mdm-2 following incomplete cerebral ischemia in rats within a period of 28 days from the insult. 72 fasted male Sprague-Dawley rats (400 g) were randomly assigned to one of the following groups: Group 1...

  7. A combined regimen of gossypol plus methyltestosterone and ethinylestradiol as a contraceptive induces germ cell apoptosis and expression of its related genes in rats.

    Science.gov (United States)

    Cui, Guang-Hui; Xu, Zeng-Lu; Yang, Zhan-Jun; Xu, Yuan-Yuan; Xue, She-Pu

    2004-10-01

    Attempts to develop gossypol and steroidal hormones alone as a male contraceptive have been tested for many years; however, both caused undesirable side effects that have prevented their acceptance. In this study, we formulated a regimen of combined gossypol at a low dose of 12 mg/kg or a high dose of 50 mg/kg plus methyltestosterone 20 mg/kg and ethinylestradiol 100 g/kg daily (12 mg G+H and 50 mg G+H) administered for 6 weeks in adult rats. The possible roles of germ cell apoptosis and related genes expression were studied by techniques of TdT-mediated dUTP nick end-labeling (TUNEL), agarose gel electrophoresis of low-molecular-weight DNA, in situ hybridization and reverse transcription-polymerase chain reaction detection. Results showed that germ cell apoptosis and related genes expression were significantly induced after combined drug administration. The apoptosis index increased 3.86- and 9.65-fold in the 12-mg and 50-mg G+H-treated groups, respectively, as compared to the control group. DNA ladder formation on the agarose gel further validated the findings of TUNEL-stained apoptotic cells. The apoptosis-related genes fas mRNA expression levels increased 0.44- and 1.39-fold, bax mRNA 0.74- and 2.56-fold, caspase-3 mRNA 0.60- and 1.29-fold, and caspase-9 mRNA 2.50- and 4.08-fold, respectively, in the 12-mg and 50-mg G+H-treated groups vs. the control group. These results indicated that our drug regimen applied as a contraceptive could induce rat germ cell apoptosis. The apoptotic process involved fas system, bax and caspase family genes and the apoptotic extent and cell types were gossypol dose-dependent.

  8. Dendrosomal curcumin nanoformulation modulate apoptosis-related genes and protein expression in hepatocarcinoma cell lines.

    Science.gov (United States)

    Montazeri, Maryam; Sadeghizadeh, Majid; Pilehvar-Soltanahmadi, Yones; Zarghami, Faraz; Khodi, Samaneh; Mohaghegh, Mina; Sadeghzadeh, Hadi; Zarghami, Nosratollah

    2016-07-25

    The side-effects observed in conventional therapies have made them unpromising in curing Hepatocellular carcinoma; therefore, developing novel treatments can be an overwhelming significance. One of such novel agents is curcumin which can induce apoptosis in various cancerous cells, however, its poor solubility is restricted its application. To overcome this issue, this paper employed dendrosomal curcumin (DNC) was employed to in prevent hepatocarcinoma in both RNA and protein levels. Hepatocarcinoma cells, p53 wild-type HepG2 and p53 mutant Huh7, were treated with DNC and investigated for toxicity study using MTT assay. Cell cycle distribution and apoptosis were analyzed using Flow-cytometry and Annexin-V-FLUOS/PI staining. Real-time PCR and Western blot were employed to analyze p53, BAX, Bcl-2, p21 and Noxa in DNC-treated cells. DNC inhibited the growth in the form of time-dependent manner, while the carrier alone was not toxic to the cell. Flow-cytometry data showed the constant concentration of 20μM DNC during the time significantly increases cell population in SubG1 phase. Annexin-V-PI test showed curcumin-induced apoptosis was enhanced in Huh7 as well as HepG2, compared to untreated cells. Followed by treatment, mRNA expression of p21, BAX, and Noxa increased, while the expression of Bcl-2 decreased, and unlike HepG2, Huh7 showed down-regulation of p53. In summary, DNC-treated hepatocellular carcinoma cells undergo apoptosis by changing the expression of genes involved in the apoptosis and proliferation processes. These findings suggest that DNC, as a plant-originated therapeutic agent, could be applied in cancer treatment. PMID:27234697

  9. Effect of Thyrotropin Releasing Hormone (TRH on Gene Expressions in Rat Pancreas: Approach by Microarray Hybridization

    Directory of Open Access Journals (Sweden)

    Luo LG

    2004-07-01

    Full Text Available CONTEXT: Thyrotropin releasing hormone (TRH, originally identified as a hypothalamic hormone, expresses in the pancreas. The effects of TRH such as, inhibiting amylase secretion in rats through a direct effect on acinar cells, enhancing basal glucagon secretion from isolated perfused rat pancreas, and potentiating glucose-stimulated insulin secretion in perfused rat islets and insulin-secreting clonal beta-cell lines, suggest that TRH may play a role in pancreas. TRH also enlarged pancreas and increased pancreatic DNA content but deletion of TRH gene expression caused hyperglycemia in mice, suggesting that TRH may play a critical role in pancreatic development; however, the biological mechanisms of TRH in the adult pancreas remains unclear. OBJECTIVES: This study explored the effect of TRH on rat pancreas. SUBJECTS: Four male-Sprague-Dawley-rats (200-250 g were given 10 microg/kg BW of TRH intraperitoneally on 1st and 3rd day and sacrificed on 7th day. Four same-strain rats without TRH injection served as controls. MAIN OUTCOME MEASURES: Wet pancreatic weights were measured. Pancreatic tissues were homogenized and extracted. The insulin levels of the extracts were measured by ELISA. Total RNA from the pancreases were fluorescently labeled and hybridized to microarray with 1,081 spot genes. RESULTS: TRH increased pancreatic wet weight and insulin contents. About 75% of the 1,081 genes were detected in the pancreas. TRH regulated up 99 genes and down 76 genes. The administration of TRH induced various types of gene expressions, such as G-protein coupled receptors (GPCR and signal transduction related genes (GPCR kinase 4, transducin beta subunit 5, arrestin beta1MAPK3, MAPK5, c-Src kinase, PKCs, PI3 kinase, growth factors (PDGF-B, IGF-2, IL-18, IGF-1, IL-2, IL-6, endothelin-1 and apoptotic factors (Bcl2, BAD, Bax. CONCLUSION: Reprogramming of transcriptome may be a way for TRH-regulation of pancreatic cellular functions.

  10. Inhibition of hepatocelluar carcinoma MAT2A and MAT2beta gene expressions by single and dual small interfering RNA

    Directory of Open Access Journals (Sweden)

    Sun Quan

    2008-11-01

    Full Text Available Abstract RNA interference (RNAi has been successfully applied in suppression of hepatic cancer genes. In hepatocelluar carcinoma cell, one methionine adenosyltransferase (MAT isozyme, MATII was found to have two catalytic subunits which were encoded by MAT2A and MAT2β respectively. During tumorigeness of hepatocelluar carcinoma, expressions of the two genes were discovered to be increased combining with a switch of MAT (form MATI to MATII, To figure out the role played by MATII in hepatic cancer, In this study, for the first time we established a dual small interfering RNA (siRNA expression system, which could simultaneously express two different siRNA molecules specifically targeting two genes. To test the effectiveness of this system, we applied this approach to express simultaneously two different siRNA duplexes that specifically target MAT2A and MAT2β genes of hepatocelluar carcinoma respectively in HepG2 cell. Results indicated that dual siRNA could simultaneously inhibit the expression of MAT2A and MAT2β gene by 89.5% and 97.8% respectively, In addition, dual siRNA molecules were able to significantly suppress growth of hepatocelluar carcinoma cell in vitro as well as induce apoptosis which was involved in arrest cell cycle at the G1/S checkpoint and the expressions of p21, p27 and Bax.

  11. Validation of the Antiproliferative Effects of Organic Extracts from the Green Husk of Juglans regia L. on PC-3 Human Prostate Cancer Cells by Assessment of Apoptosis-Related Genes

    Directory of Open Access Journals (Sweden)

    Ali A. Alshatwi

    2012-01-01

    Full Text Available With the increased use of plant-based cancer chemotherapy, exploring the antiproliferative effects of phytochemicals for anticancer drug design has gained considerable attention worldwide. This study was undertaken to investigate the effect of walnut green husk extracts on cell proliferation and to determine the possible molecular mechanism of extract-induced cell death by quantifying the expression of Bcl-2, Bax, caspases-3, and Tp53. PC-3 human prostate cancer cells. In this study, we found that green husk extracts suppressed proliferation and induced apoptosis in a dose- and time-dependent manner by modulating expression of apoptosis-related genes. This involved DNA fragmentation (determined by TUNEL assay and significant changes in levels of mRNA and the expression of corresponding proteins. An increase in expressions of Bax, caspase-3, and tp53 genes and their corresponding proteins was detected using real-time PCR and western blot analysis in PC-3 cells treated with the green husk organic extracts. In contrast, Bcl2 expression was downregulated after exposure to the extracts. Our data suggest the presence of bioactive compound(s in walnut green husks that are capable of killing prostate carcinoma cells by inducing apoptosis and that the husks are a candidate source of anticancer drugs.

  12. In vitro effects of relaxin on gene expression in porcine cumulus-oocyte complexes and developing embryos

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    Willard Scott T

    2011-01-01

    Full Text Available Abstract Background Relaxin hormone peptide is found in porcine follicular and utero-tubal fluids, but its possible actions during early embryo development are still undetermined. Here, we investigated the effects of porcine relaxin during oocyte maturation and embryo development, and gene expression in the pig. Methods Immature cumulus-oocyte complexes (COCs were obtained from ovarian follicles of sows. In experiment 1, COCs were matured in the presence of 0, 20, or 40 ng relaxin/ml, or 10% (v/v porcine follicular fluid. In experiment 2, COCs were in vitro matured, fertilized and resulting embryos were cultured in the presence of 0, 20, or 40 ng relaxin/ml. In experiment 3, COCs were matured in the presence of 40 ng relaxin/ml, fertilized and zygotes were cultured as indicated in experiment 2. We evaluated the proportions of matured oocytes in experiment 1, cleaved and blastocysts on Day 2 and Day 7 post insemination in all experiments. The total cell number of blastocysts was also evaluated. In parallel, transcription levels of both relaxin and its receptors (RXFP1 and RXFP2, as well as a pro- (Bax and anti- (Bcl2-like 1 apoptotic-related genes were determined. All data were analyzed by ANOVA and significant differences were fixed for P Results In experiment 1, relaxin significantly increased the proportions of matured oocytes and cleaved embryos, as well as the expression level of RXFP2 mRNA compared to RXFP1 (P Conclusions Exogenous relaxin influences its own receptors expression, improves oocyte nuclear maturation. Its beneficial effect on total cell number of blastocysts appears to be through a Bcl2-like1/Bax-independent mechanism.

  13. Effect of single prolonged stress on behavior and expression of Bcl-2 and Bax in the rat hippocampus%单一连续应激对大鼠行为学和海马Bcl-2、Bax表达的影响

    Institute of Scientific and Technical Information of China (English)

    徐爱军; 耿菲; 王海涛

    2012-01-01

    Objective To observe the behavior change and the expression of Bcl-2 and Bax in the hippocampus of rats after treated with single-prolonged stress. Method Wistar rats were divided randomly into control group and stress group,and the stress rats were treated with single-prolonged stress. The behavior changes of rats were observed with open-field test, reject grip test and Morries water mace test. Hydrocortisone level in rats blood serum was determined with chemi-luminescence. Immunohistochemistry was used to detect the expressions of Bax and Bal-2 in the hippocampal neurons. . Results Morris water mace test showed that escape latency of control group was 5. 632s ± 1. 065s and stress group was 20. 762s ±3.236s(t = 8.934,P<0.01). Hydrocortisone level of control group was 1.25μg/dl ±0. 12μg/dl and model was 0. 58μg/dl ±0.09μg/dl(t =8.075 ,P <0.01). The ratio of Bcl-2/Bax in control group was 0.904 ±0.109, while the stress group was 1.440 ±0.091(t = 5.458,P <0. 01). Conclusion The neuronal apoptosis in hippocampus may play a role in posttraumatic stress disorder. The increasing of Bcl-2 and Bax protein and the change of Bcl-2/Bax ratio may be the mechanism during single-prolonged stress.%目的 检测单一连续应激(single prolonged stress,SPS)对大鼠行为学和海马神经细胞Bax、Bcl-2表达的影响.方法 将Wistar大鼠分为对照组和应激组.应激组给予SPS应激,采用旷场实验、拒俘反应实验、Morries水迷宫实验等观察大鼠情感行为改变,采用化学发光法测定血清皮质醇浓度,应用免疫组织化学方法检测海马神经元Bcl-2、Bax的表达.结果 对照组水迷宫实验逃避潜伏期为5.632s±1.065s,应激组为20.762s±3.236s(t=9.932,P<0.01);对照组血清皮质醇浓度为1.25 μg/dl ±0.12μg/dl,应激组为0.58μg/dl±0.09μg/dl(t=7.340,P<0.01).应激组海马Bcl-2、Bax表达升高,Bcl-2/Bax上调.结论 SPS应激能使大鼠表现出创伤后应激障碍行

  14. 30例自然流产患者绒毛滋养细胞凋亡及调控蛋白Bcl-2、Bax的表达研究%Study on apoptosis and expression of modulin Bcl-2, Bax in villus syncytiotrophoblast cells in 30 patients with spontaneous abortion

    Institute of Scientific and Technical Information of China (English)

    郭冬瑾; 林秀玲; 樊柳宜

    2012-01-01

    目的 通过观察绒毛合体滋养细胞凋亡以及凋亡调控蛋白Bcl-2和Bax在自然流产患者中的表达,探讨细胞凋亡在自然流产的发病机制.方法 采用原位末端标记法(TUNEL)和免疫组织化学法对30例自然流产患者绒毛合体滋养细胞中调亡指数及凋亡调控蛋白Bcl-2和Bax阳性表达率进行检测,并以30例人工终止早孕的健康妇女做对照(对照组),所有数据采用SPSS16.0进行统计学分析.结果 自然流产组绒毛滋养细胞中凋亡指数为(32.45±5.87)%,明显高于对照组的(19.38±4.16)%,差异有统计学意义(P<0.01).Bax和Bcl-2在两组合体滋养细胞中的阳性率比较显示,自然流产组中Bax阳性率增高,Bcl-2阳性率下降,Bcl-2/Bax比值升高,差异均有统计学意义(P<0.01).结论 绒毛合体滋养细胞凋亡明显增加、Bax阳性率增高和Bcl-2阳性率下降在自然流产中产生重要作用.%Objective To observe the apoptosis and expression of modulin Bcl-2, Bax in villus syncytiotrophoblast cells in patients with spontaneous abortion, and to investigate the pathogenesis of apoptosis in spontaneous abortion. Methods TdT-mediated dUTP nick end labeling (TUNEL) and immunohistochemistry were respectively used in 30 patients with spontaneous abortion (the study group) and 30 individuals with artificial abortion (the control group). The apoptosis index, the positive expression rates of Bax and Bcl-2 in villus syncytiotrophoblast cells were detected. All the data were analyzed by SPSS 16.0. Results The apoptosis index of syncytiotrophoblast cells in the study group was (32.45±5.87)%, significantly higher than (19.38±4.16)% in the control group (P<0.01). The positive expression rate of Bax in the study group was significantly higher than in the control group, while that of Bcl-2 was significantly lower and Bcl-2/Bax ratio was significantly higher (P<0.01). Conclusion Intensive apoptosis, the increasing positive expression rate of Bax, and

  15. Gene gymnastics

    Science.gov (United States)

    Vijayachandran, Lakshmi S; Thimiri Govinda Raj, Deepak B; Edelweiss, Evelina; Gupta, Kapil; Maier, Josef; Gordeliy, Valentin; Fitzgerald, Daniel J; Berger, Imre

    2013-01-01

    Most essential activities in eukaryotic cells are catalyzed by large multiprotein assemblies containing up to ten or more interlocking subunits. The vast majority of these protein complexes are not easily accessible for high resolution studies aimed at unlocking their mechanisms, due to their low cellular abundance and high heterogeneity. Recombinant overproduction can resolve this bottleneck and baculovirus expression vector systems (BEVS) have emerged as particularly powerful tools for the provision of eukaryotic multiprotein complexes in high quality and quantity. Recently, synthetic biology approaches have begun to make their mark in improving existing BEVS reagents by de novo design of streamlined transfer plasmids and by engineering the baculovirus genome. Here we present OmniBac, comprising new custom designed reagents that further facilitate the integration of heterologous genes into the baculovirus genome for multiprotein expression. Based on comparative genome analysis and data mining, we herein present a blueprint to custom design and engineer the entire baculovirus genome for optimized production properties using a bottom-up synthetic biology approach. PMID:23328086

  16. Conserved regulation of p53 network dosage by microRNA-125b occurs through evolving miRNA-target gene pairs.

    Directory of Open Access Journals (Sweden)

    Minh T N Le

    2011-09-01

    Full Text Available MicroRNAs regulate networks of genes to orchestrate cellular functions. MiR-125b, the vertebrate homologue of the Caenorhabditis elegans microRNA lin-4, has been implicated in the regulation of neural and hematopoietic stem cell homeostasis, analogous to how lin-4 regulates stem cells in C. elegans. Depending on the cell context, miR-125b has been proposed to regulate both apoptosis and proliferation. Because the p53 network is a central regulator of both apoptosis and proliferation, the dual roles of miR-125b raise the question of what genes in the p53 network might be regulated by miR-125b. By using a gain- and loss-of-function screen for miR-125b targets in humans, mice, and zebrafish and by validating these targets with the luciferase assay and a novel miRNA pull-down assay, we demonstrate that miR-125b directly represses 20 novel targets in the p53 network. These targets include both apoptosis regulators like Bak1, Igfbp3, Itch, Puma, Prkra, Tp53inp1, Tp53, Zac1, and also cell-cycle regulators like cyclin C, Cdc25c, Cdkn2c, Edn1, Ppp1ca, Sel1l, in the p53 network. We found that, although each miRNA-target pair was seldom conserved, miR-125b regulation of the p53 pathway is conserved at the network level. Our results lead us to propose that miR-125b buffers and fine-tunes p53 network activity by regulating the dose of both proliferative and apoptotic regulators, with implications for tissue stem cell homeostasis and oncogenesis.

  17. Principles of gene therapy

    OpenAIRE

    Mammen Biju; Ramakrishnan T; Sudhakar Uma; Vijayalakshmi

    2007-01-01

    Genes are specific sequences of bases that encode instructions to make proteins. When genes are altered so that encoded proteins are unable to carry out their normal functions, genetic disorders can result. Gene therapy is designed to introduce genetic material into cells to compensate for abnormal genes or to make a beneficial protein. This article reviews the fundamentals in gene therapy and its various modes of administration with an insight into the role of gene therapy in Periodontics an...

  18. 灯盏花注射液抗新生鼠缺氧缺血性脑损伤的作用及对Bcl-2、Bax蛋白表达的影响%The anti-injury effect of Breviscapine injection on the hypoxic ischemic brain damage of neonatal rats and the expression of Bcl-2 and Bax

    Institute of Scientific and Technical Information of China (English)

    张明艳; 范淑娟; 李利平; 武变瑛; 王宇

    2011-01-01

    目的:观察灯盏花注射液对新生大鼠缺氧缺血脑损伤(HIBD)的保护作用及对Bcl-2、Bax蛋白表达的影响.方法:采用新生7日龄SD大鼠缺氧缺血性脑损伤模型,设立假手术组、缺氧缺血脑损伤模型组、灯盏花注射液治疗大、中、小剂量组、无菌注射用水对照组.采用硫堇染色、免疫组织化学染色的方法测定各组大鼠海马CA1区神经元密度、组织学分级及凋亡相关基因Bcl-2、Bax蛋白表达情况,并计数各时间点Bcl-2、Bax免疫阳性细胞数目及测定积分光密度值.结果:假手术组,大鼠海马CA1区无锥体细胞缺失,未见明显免疫阳性细胞.与假手术组比较,缺氧缺血脑损伤模型组、无菌注射用水对照组Bcl-2、Bax蛋白表达均于3 d时达到高峰(与其余各时间点比较差别有显著意义P<0.05),神经元密度明显降低,组织学分级显著增高,积分光密度值增加.灯盏花治疗组,与无菌注射用水对照组比较,Bcl-2蛋白表达进一步增加,积分光密度值增加;而Bax蛋白表达则减少,积分光密度值降低;神经元密度显著高于对照组,组织学分级明显降低.结论:灯盏花注射液可能是通过上调Bcl-2表达,抑制Bax表达,减轻缺氧缺血引起的神经元凋亡及迟发性神经元死亡.%Objective: To observe the protective effect of Breviscapine injection on the hypoxic ischemic brain damage of neonatal rats, and the expression of Bcl-2 and Bax. Methods: In this experiment 7-day-old newborn rat with hypoxic-ischemic brain damage model was used and divided into four groups: sham group, model group, control with sterile water for injection group and Breviscapine injection group. Breviscapine injection group was divided into large, medium, and small doses. Used thionin staining and immunohistochemical staining to assay the neuronal density, histological grade, and the expresssion of Bcl-2 and Bax protein in the CA1 hippocampus of each group , the number of

  19. The regulation of thermal stress induced apoptosis in corals reveals high similarities in gene expression and function to higher animals

    Science.gov (United States)

    Kvitt, Hagit; Rosenfeld, Hanna; Tchernov, Dan

    2016-01-01

    Recent studies suggest that controlled apoptotic response provides an essential mechanism, enabling corals to respond to global warming and ocean acidification. However, the molecules involved and their functions are still unclear. To better characterize the apoptotic response in basal metazoans, we studied the expression profiles of selected genes that encode for putative pro- and anti-apoptotic mediators in the coral Stylophora pistillata under thermal stress and bleaching conditions. Upon thermal stress, as attested by the elevation of the heat-shock protein gene HSP70’s mRNA levels, the expression of all studied genes, including caspase, Bcl-2, Bax, APAF-1 and BI-1, peaked at 6–24 h of thermal stress (hts) and declined at 72 hts. Adversely, the expression levels of the survivin gene showed a shifted pattern, with elevation at 48–72 hts and a return to basal levels at 168 hts. Overall, we show the quantitative anti-apoptotic traits of the coral Bcl-2 protein, which resemble those of its mammalian counterpart. Altogether, our results highlight the similarities between apoptotic networks operating in simple metazoans and in higher animals and clearly demonstrate the activation of pro-cell survival regulators at early stages of the apoptotic response, contributing to the decline of apoptosis and the acclimation to chronic stress. PMID:27460544

  20. The regulation of thermal stress induced apoptosis in corals reveals high similarities in gene expression and function to higher animals

    Science.gov (United States)

    Kvitt, Hagit; Rosenfeld, Hanna; Tchernov, Dan

    2016-07-01

    Recent studies suggest that controlled apoptotic response provides an essential mechanism, enabling corals to respond to global warming and ocean acidification. However, the molecules involved and their functions are still unclear. To better characterize the apoptotic response in basal metazoans, we studied the expression profiles of selected genes that encode for putative pro- and anti-apoptotic mediators in the coral Stylophora pistillata under thermal stress and bleaching conditions. Upon thermal stress, as attested by the elevation of the heat-shock protein gene HSP70’s mRNA levels, the expression of all studied genes, including caspase, Bcl-2, Bax, APAF-1 and BI-1, peaked at 6–24 h of thermal stress (hts) and declined at 72 hts. Adversely, the expression levels of the survivin gene showed a shifted pattern, with elevation at 48–72 hts and a return to basal levels at 168 hts. Overall, we show the quantitative anti-apoptotic traits of the coral Bcl-2 protein, which resemble those of its mammalian counterpart. Altogether, our results highlight the similarities between apoptotic networks operating in simple metazoans and in higher animals and clearly demonstrate the activation of pro-cell survival regulators at early stages of the apoptotic response, contributing to the decline of apoptosis and the acclimation to chronic stress.

  1. The regulation of thermal stress induced apoptosis in corals reveals high similarities in gene expression and function to higher animals.

    Science.gov (United States)

    Kvitt, Hagit; Rosenfeld, Hanna; Tchernov, Dan

    2016-01-01

    Recent studies suggest that controlled apoptotic response provides an essential mechanism, enabling corals to respond to global warming and ocean acidification. However, the molecules involved and their functions are still unclear. To better characterize the apoptotic response in basal metazoans, we studied the expression profiles of selected genes that encode for putative pro- and anti-apoptotic mediators in the coral Stylophora pistillata under thermal stress and bleaching conditions. Upon thermal stress, as attested by the elevation of the heat-shock protein gene HSP70's mRNA levels, the expression of all studied genes, including caspase, Bcl-2, Bax, APAF-1 and BI-1, peaked at 6-24 h of thermal stress (hts) and declined at 72 hts. Adversely, the expression levels of the survivin gene showed a shifted pattern, with elevation at 48-72 hts and a return to basal levels at 168 hts. Overall, we show the quantitative anti-apoptotic traits of the coral Bcl-2 protein, which resemble those of its mammalian counterpart. Altogether, our results highlight the similarities between apoptotic networks operating in simple metazoans and in higher animals and clearly demonstrate the activation of pro-cell survival regulators at early stages of the apoptotic response, contributing to the decline of apoptosis and the acclimation to chronic stress. PMID:27460544

  2. Organization of immunoglobulin genes.

    Science.gov (United States)

    Tonegawa, S; Brack, C; Hozumi, N; Pirrotta, V

    1978-01-01

    The nucleotide-sequence determination of a cloned, embryonic Vlambda gene directly demonstrated that V genes are separate from a corresponding C gene in embryonic cells. Analysis by restriction enzymes of total cellular DNA from various sources strongly suggested that the two separate immunoglobulin genes become continuous during differentiation of B lymphocytes. There seems to be a strict correlation between the joining event and activation of the joined genes. Cloning of more immunoglobulin genes from embryo and plasma cells will not only provide direct demonstration of such a gene-joining event but also help in the elucidation of a possible relationship of the event to gene activation mechanisms.

  3. Imaging gene expression in gene therapy

    International Nuclear Information System (INIS)

    Full text. Gene therapy can be used to introduce new genes, or to supplement the function of indigenous genes. At the present time, however, there is non-invasive test to demonstrate efficacy of the gene transfer and expression processes. It has been postulated that scintigraphic imaging can offer unique information on both the site at which the transferred gene is expressed, and the degree of expression, both of which are critical issue for safety and clinical efficacy. Many current studies are based on 'suicide gene therapy' of cancer. Cells modified to express these genes commit metabolic suicide in the presence of an enzyme encoded by the transferred gene and a specifically-convertible pro drug. Pro drug metabolism can lead to selective metabolic trapping, required for scintigraphy. Herpes simplex virus type-1 thymidine kinase (H S V-1 t k+) has been use for 'suicide' in vivo tumor gene therapy. It has been proposed that radiolabelled nucleosides can be used as radiopharmaceuticals to detect H S V-1 t k+ gene expression where the H S V-1 t k+ gene serves a reporter or therapeutic function. Animal gene therapy models have been studied using purine-([18 F]F H P G; [18 F]-A C V), and pyrimidine- ([123/131 I]I V R F U; [124/131I]) antiviral nucleosides. Principles of gene therapy and gene therapy imaging will be reviewed and experimental data for [123/131I]I V R F U imaging with the H S V-1 t k+ reporter gene will be presented

  4. Imaging gene expression in gene therapy

    Energy Technology Data Exchange (ETDEWEB)

    Wiebe, Leonard I. [Alberta Univ., Edmonton (Canada). Noujaim Institute for Pharmaceutical Oncology Research

    1997-12-31

    Full text. Gene therapy can be used to introduce new genes, or to supplement the function of indigenous genes. At the present time, however, there is non-invasive test to demonstrate efficacy of the gene transfer and expression processes. It has been postulated that scintigraphic imaging can offer unique information on both the site at which the transferred gene is expressed, and the degree of expression, both of which are critical issue for safety and clinical efficacy. Many current studies are based on `suicide gene therapy` of cancer. Cells modified to express these genes commit metabolic suicide in the presence of an enzyme encoded by the transferred gene and a specifically-convertible pro drug. Pro drug metabolism can lead to selective metabolic trapping, required for scintigraphy. Herpes simplex virus type-1 thymidine kinase (H S V-1 t k{sup +}) has been use for `suicide` in vivo tumor gene therapy. It has been proposed that radiolabelled nucleosides can be used as radiopharmaceuticals to detect H S V-1 t k{sup +} gene expression where the H S V-1 t k{sup +} gene serves a reporter or therapeutic function. Animal gene therapy models have been studied using purine-([{sup 18} F]F H P G; [{sup 18} F]-A C V), and pyrimidine- ([{sup 123}/{sup 131} I]I V R F U; [{sup 124}/{sup 131I}]) antiviral nucleosides. Principles of gene therapy and gene therapy imaging will be reviewed and experimental data for [{sup 123}/{sup 131I}]I V R F U imaging with the H S V-1 t k{sup +} reporter gene will be presented

  5. Altered expression of apoptotic genes in response to OCT4B1 suppression in human tumor cell lines.

    Science.gov (United States)

    Mirzaei, Mohammad Reza; Najafi, Ali; Arababadi, Mohammad Kazemi; Asadi, Malek Hosein; Mowla, Seyed Javad

    2014-10-01

    OCT4B1 is a newly discovered spliced variant of OCT4 which is primarily expressed in pluripotent and tumor cells. Based on our previous studies, OCT4B1 is significantly overexpressed in tumors, where it endows an anti-apoptotic property to tumor cells. However, the mechanism by which OCT4B1 regulates the apoptotic pathway is not yet elucidated. Here, we investigated the effects of OCT4B1 suppression on the expression alteration of 84 genes involved in apoptotic pathway. The AGS (gastric adenocarcinoma), 5637 (bladder tumor), and U-87MG (brain tumor) cell lines were transfected with OCT4B1 or irrelevant siRNAs. The expression level of apoptotic genes was then quantified using a human apoptosis panel-PCR kit. Our data revealed an almost similar pattern of alteration in the expression profile of apoptotic genes in all three studied cell lines, following OCT4B1 suppression. In general, the expression of more than 54 apoptotic genes (64 % of arrayed genes) showed significant changes. Among these, some up-regulated (CIDEA, CIDEB, TNFRSF1A, TNFRSF21, TNFRSF11B, TNFRSF10B, and CASP7) and down-regulated (BCL2, BCL2L11, TP73, TP53, BAD, TRAF3, TRAF2, BRAF, BNIP3L, BFAR, and BAX) genes had on average more than tenfold gene expression alteration in all three examined cell lines. With some minor exceptions, suppression of OCT4B1 caused upregulation of pro-apoptotic and down-regulation of anti-apoptotic genes in transfected tumor cells. Uncovering OCT4B1 down-stream targets could further elucidate its part in tumorigenesis, and could lead to finding a new approach to combat cancer, based on targeting OCT4B1. PMID:25008565

  6. Identifying Gene Interaction Enrichment for Gene Expression Data

    OpenAIRE

    Jigang Zhang; Jian Li; Hong-Wen Deng

    2009-01-01

    Gene set analysis allows the inclusion of knowledge from established gene sets, such as gene pathways, and potentially improves the power of detecting differentially expressed genes. However, conventional methods of gene set analysis focus on gene marginal effects in a gene set, and ignore gene interactions which may contribute to complex human diseases. In this study, we propose a method of gene interaction enrichment analysis, which incorporates knowledge of predefined gene sets (e.g. gene ...

  7. The role' of Fas/FasL in radiation induced apoptosis in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sung Hee; Seong, Jin Sil [College of Medicine, Yonsei Univ., Seoul (Korea, Republic of)

    2003-09-01

    The interaction of the Fas: Fas ligand has been recognized to play an important role in radiation-induced apoptosis. The purpose of this study was to investigate the role of Fas and Fas ligand mutations, in radiation-induced apoptosis in vivo. Mice with a mutation in the Fas (C57BL6J-Fas{sup lpr}) and its normal control (C57BU 6J) and the Fas ligand (C3H/HeJ-Fas{sup gId}) and its normal control (C3H/HeJ), were used in this study. Eight-week old male mice were given whole body radiation. After irradiation, the mice were killed at various time intervals, and their spleens collected. Tissue sample was stained with hematoxylin-eosin, and the numbers of apoptotic cells scored. The regulating molecules of apoptosis including the p53, Bcl-2, Bax, Bcl-X{sup L} and Bcl-X{sup s} genes were also analyzed by Western blotting. With 2.5 Gy and 10 Gy of irradiation, the levels of apoptosis were lower in the C57BL/6J-Fas{sup lpr} and C3H/HeJ-Fas{sup gld} mice than in the control mice (p < 0.05). With the expression of apoptosis regulating molecules, the Bax was increased in both the C57BL/6J and C3H/HeJ mice in response to radiation; the peak levels of Bax in the C57BL/6J and C3H/HeJ were 3 and 3.3-fold higher after 8hr, respectively. However the Bax was not increased in either the C57BL/6J-Fas{sup lpr} or C3H/HeJ-Fas{sup gld} mice. The p53, Bcl-X{sup L}, Bcl-X{sub s} and Bcl-2 showed no significant changes in the C57BL/6J-Fas{sup lpr}, C3H/HeJ-Fas{sup gld}, C57BL/6J and C3H/HeJ mice. The levels of radiation-induced apoptosis were lower in the /pr and gld, than the control mice, which seemed to be related to the level of Bax activation due to the radiation in the /pr and gld mice, This result suggests that Fas/Fas L plays an important role in radiation-induced apoptosis in vivo.

  8. Expression of Inflammatory and Cell Death Program Genes and Comet DNA Damage Assay Induced by Escherichia coli in Layer Hens.

    Science.gov (United States)

    Mehaisen, Gamal M K; Eshak, Mariam G; El Sabry, M I; Abass, Ahmed O

    2016-01-01

    Modern methods of industrial poultry and egg production systems involve stressful practices that stimulate Escherichia coli (E. coli) activity causing endotoxic shock. This investigation was conducted to evaluate the expression of pro-inflammatory cytokines and cell death program genes and DNA damage induced by E. coli in the brain and liver tissues of laying hens. A total of two hundred and ten H&N brown layer hens with 20 week age, were used in this research. First, preliminary experiments were designed (60 hens in total) to establish the optimal exposure dose of E. coli and to determine the nearest time of notable response to be used in the remainder studies of this research. At 35-wk of age, 150 hens were randomly assigned into 2 groups with 3 replicates of 25 birds each; the first group was injected in the brachial wing vein with 107 E. coli colony/hen, while the second group was injected with saline and served as a control. The body temperature and plasma corticosterone concentration were measured 3 hr after injection. Specimens of liver and brain were obtained from each group and the gene expression of p38 mitogen-activated protein kinase, interlukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α), Bax, and caspase-3 genes were measured by quantitative real-time PCR. DNA damage in the brain and liver tissues were also measured by comet assay. Hens treated with E. coli showed significant (Phens injected with E. coli showed an increase in DNA damage in the brain and liver cells (P<0.05). These results were synchronized with activating cell death program since our data showed significant high expression of Bax gene by 2.8- and 2.7-fold and caspase-3 gene by 2.5- and 2.7-fold in the brain and liver tissues of infected chickens, respectively (P<0.05). In conclusion, the current study indicates that E. coli injection induces inflammatory physiological response and triggers cell death program in the brain and liver. Our results provide more understanding to

  9. Expression of Inflammatory and Cell Death Program Genes and Comet DNA Damage Assay Induced by Escherichia coli in Layer Hens

    Science.gov (United States)

    Mehaisen, Gamal M. K.; Eshak, Mariam G.; El Sabry, M. I.; Abass, Ahmed O.

    2016-01-01

    Modern methods of industrial poultry and egg production systems involve stressful practices that stimulate Escherichia coli (E. coli) activity causing endotoxic shock. This investigation was conducted to evaluate the expression of pro-inflammatory cytokines and cell death program genes and DNA damage induced by E. coli in the brain and liver tissues of laying hens. A total of two hundred and ten H&N brown layer hens with 20 week age, were used in this research. First, preliminary experiments were designed (60 hens in total) to establish the optimal exposure dose of E. coli and to determine the nearest time of notable response to be used in the remainder studies of this research. At 35-wk of age, 150 hens were randomly assigned into 2 groups with 3 replicates of 25 birds each; the first group was injected in the brachial wing vein with 107 E. coli colony/hen, while the second group was injected with saline and served as a control. The body temperature and plasma corticosterone concentration were measured 3 hr after injection. Specimens of liver and brain were obtained from each group and the gene expression of p38 mitogen-activated protein kinase, interlukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α), Bax, and caspase-3 genes were measured by quantitative real-time PCR. DNA damage in the brain and liver tissues were also measured by comet assay. Hens treated with E. coli showed significant (Phens injected with E. coli showed an increase in DNA damage in the brain and liver cells (P<0.05). These results were synchronized with activating cell death program since our data showed significant high expression of Bax gene by 2.8- and 2.7-fold and caspase-3 gene by 2.5- and 2.7-fold in the brain and liver tissues of infected chickens, respectively (P<0.05). In conclusion, the current study indicates that E. coli injection induces inflammatory physiological response and triggers cell death program in the brain and liver. Our results provide more understanding to

  10. Autism and Genes

    Science.gov (United States)

    National Institutes of Health, 2005

    2005-01-01

    This document defines and discusses autism and how genes play a role in the condition. Answers to the following questions are covered: (1) What are genes? (2) What is autism? (3) What causes autism? (4) Why study genes to learn about autism? (5) How do researchers look for the genes involved in autism? (screen the whole genome; conduct cytogenetic…

  11. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.;

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes we...

  12. FK506对大鼠面神经损伤后面运动神经元凋亡及bcl-2,bax表达的影响%Effects of FK506 on the Apoptosis and bcl-2, bax Expression of Rat Facial Motor Neurons after Facial Nerve Injury

    Institute of Scientific and Technical Information of China (English)

    惠莲; 刘凤啸; 袁婧; 姜学钧; 任重

    2013-01-01

    目的 研究FK506对大鼠面神经损伤后运动神经元凋亡及bcl-2和bax表达的影响.方法 将40只Wistar大鼠随机分为实验组和对照组,制作大鼠单侧面神经总干切断模型,实验组每日腹腔注射FK506注射液,对照组给予相同剂量的盐水,在术后各时间点,通过Nissl染色观测面神经核神经元的形态学变化;TUNEL检测细胞凋亡;免疫组化检测bcl-2和bax表达的变化.结果 面神经损伤后,FK506组与盐水组相比较,FK506组运动神经元凋亡明显减少,bcl-2表达增加,bax表达降低.结论 FK506有助于增加面神经损伤后运动神经元bcl-2的表达、降低bax表达,抑制细胞凋亡,为药物在神经损伤疾病的临床应用提供了实验依据.%Objective To investigate effects of FKS06 on the apoptosis and the expression of bcl-2 and bax of rat facial motor neurons after facial nerve injury.Methods A total of 40 Wistar rats were randomly divided into experimental and control group.Facial nerve injury model was established by transecting facial nerves at its stylomastoid foramen,and then the experimental group and the control group were treated with FK506(FK506-treated group) and normal saline(saline control group)by intraperitoneal injection,respectively.The morphology of facial neurons were observed under light microscope at different time points after injury;the apoptotic cells of injured facial motor neurons were detected by TUNEL staining;the expression of bcl-2 and bax were evaluated by immunohistochemistry method.Results After facial nerve transection,the apoptotic cells were significantly decreased in FK506-treated group compared with saline control group (P < 0.05).The expression level of bcl-2 in FKS06-treated group were higher than that in saline control group,and the expression level of bax in FK506-treated group were lower than the control group.Conclusion FK506 could increase the expression of bcl-2,and decrease the expression of bax,and inhibit the

  13. 光动力学调控miR143激活Bcl-2/Bax的信号路径对人宫颈癌的治疗机制%Regulation Effect of Photodynamic Therapy on Bcl-2/Bax Signal Path Activated by miR143 in Human Cervical Cancer

    Institute of Scientific and Technical Information of China (English)

    兰艳丽; 刘韵

    2016-01-01

    Objective To investigate the regulation effect of photodynamic therapy on Bcl-2/Bax signal path activated by miR143 in human cervical cancer ,and lay foundation for cervical cancer treated with photodynamic therapy along with miR 143. Methods Human HeLa cells received miR 143 interference ( miR143 group ) , photodynamic irradiation treatment ( PDT group ) and miR143 interference combined with photodynamic irradiation (PDT +miR143 group).The rate of inhibition,apoptosis and invasion were detected by CCK8,flow cytometry and Transwell model ,respectively.Meanwhile the expression levels of miR143, Bcl-2 and Bax before and after different treatments were detected by fluorescence quantitative PCR .Results The inhibition rates and apoptosis rates among the 3 groups were significantly different (P0.05).Conclusion In cervical cancer,photodynamic therapy upregulated the Bcl-2/Bax signal path activated by miR 143,and lay foundation for cervical cancer treated with photodynamic therapy along with miR 143.%目的 探讨在宫颈癌中光动力学调控对miR143激活Bcl-2/Bax的信号路径的影响,为光动力学联合miR143应用于宫颈癌的治疗机制奠定基础.方法 人宫颈癌HeLa细胞分别进行miR143干扰处理(miR143组)、光动力照射处理(PDT组)及miR143干扰联合光动力照射处理(PDT+miR143组),采用CCK8法、流式细胞术及Transwell小室侵袭模型对各组处理后HeLa细胞的抑制率、凋亡率及侵袭能力进行分析,同时采用荧光定量PCR检测不同处理前后各组细胞的miR143、Bcl-2和Bax的mRNA表达水平.结果 3组之间的细胞抑制率、凋亡率差异有统计学意义(P<0.05),PDT+miR143组细胞抑制率和凋亡率分别高于PDT组和miR143组,差异具有统计学意义(P<0.05),而PDT+miR143组穿膜细胞数显著高于miR143组和PDT组(P<0.05).处理后,3组miR143和Bcl-2 mRNA表达水平较处理前均显著升高(P<0.0001),且处理后PDT+miR143组miR143和Bcl-2 mRNA表达

  14. Expression level of bcl-2 and bax in endometrial hyperplasia glandular epithelia with mifepristone treatment%Bcl-2、Bax在米非司酮治疗前后子宫内膜单纯性增生组织中的表达

    Institute of Scientific and Technical Information of China (English)

    高燕; 章明放; 付稳; 季福水

    2011-01-01

    Objective: To explore the adjust mechanism of simple hyperplasia (SH) endometrium treated by mifepristone. Methods: Immunohistochemical SP method was used to measure the expression of bcl-2 and bax in 10 normal endometrium (5 cases proliferative endometrium and secretory endometrium respectively), 43 SH and 43 endometrium after treatment with mifepristone. Results: The expression of bcl-2 in proliferative endometrium (PE) was higher than that in secretory endometrium (SE)(P<0.05), and that in SH was higher than that in PE (P<0.05). Compared with SH, the expression of bcl-2 decreased after treatment with mifepristone (P<0.05). The expression of bax in SE was higher than that in PE (P<0.05), and that in SH was lower than in PE (P<0.05). Compared with SH, the expression of bax increased after treatment with mifepristone (P<0.05). Conclusion: Mifepristone induced bax levels, while suppressing bcl-2 levels. The ability of mifepristone probably promote cellular apoptosis, which was one of the reasons to reduce endometrial hyperplasia.%目的:探讨米非司酮对子宫内膜单纯性增生的调节机制.方法:正常子宫内膜组织10例(增生期和分泌期各5例),单纯性增生子宫内膜43例,同一患者口服3个月米非司酮(10 mg/d)治疗后的子宫内膜标本43例.采用链酶菌素-生物素(S-P)免疫组化染色方法测定bcl-2、bax的表达量采用SPSS11.5统计学软件包进行数据处理,以P<0.05为有显著性差异.结果:增生期子宫内膜bcl-2阳性信号明显高于分泌期(P<0.05),单纯性增生bcl-2表达明显高于正常增生期子宫内膜(P<0.05).与单纯性增生相比,米非司酮治疗后,子宫内膜bcl-2明显下降(P<0.05).分泌期bax表达明显高于增生期(P<0.05),单纯性增生bax表达明显低于正常增生期子宫内膜(P<0.05).与单纯性增生相比,米非司酮治疗后,子宫内膜bax明显增高(P<0.05).结论:RU486增加了子宫内膜bax的表达,减少了bcl-2的表达,可能是其

  15. Principles of gene therapy

    Directory of Open Access Journals (Sweden)

    Mammen Biju

    2007-01-01

    Full Text Available Genes are specific sequences of bases that encode instructions to make proteins. When genes are altered so that encoded proteins are unable to carry out their normal functions, genetic disorders can result. Gene therapy is designed to introduce genetic material into cells to compensate for abnormal genes or to make a beneficial protein. This article reviews the fundamentals in gene therapy and its various modes of administration with an insight into the role of gene therapy in Periodontics and future percepts and the technical and ethical issues of using gene therapy.

  16. Experimental research of RB94 gene transfection into retinoblastoma cells using ultrasound-targeted microbubble destruction.

    Science.gov (United States)

    Zheng, Min-Ming; Zhou, Xi-Yuan; Wang, Li-Ping; Wang, Zhi-Gang

    2012-06-01

    The purpose of this study was to explore the transfection of the recombinant expression plasmid pEGFP-C1/RB94 into human retinoblastoma cells (HXO-Rb44) using ultrasound-targeted microbubble destruction (UTMD). pEGFP-C1/RB94 was transfected into HXO-Rb44 in vitro by UTMD, with liposome as the positive control. After 24 to 72 h, the expression of the reporter gene enhanced green fluorescent protein (EGFP) was observed using fluorescent microscopy and flow cytometry. The cell viability of HXO-Rb44 was measured by a MTT assay. The mRNA and proteins of RB94, caspase-3 and Bax were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Moreover, the apoptosis rate and cell cycle progression of the cells were detected by flow cytometry. This study demonstrated that UTMD can enhance the transfection efficiency of RB94, which has an obvious impact on the inhibition of the growth process of retinoblastoma cells, suggesting that the combination of UTMD and RB94 compounds might be a useful tool for use in the gene therapy of retinoblastoma.

  17. Blastocyst recovery and multifactorial gene expression analysis in the wild guinea pig (Cavia aperea).

    Science.gov (United States)

    Hribal, Romy; Guenther, Anja; Rübensam, Kathrin; Jewgenow, Katarina

    2016-09-15

    The expression of specific developmentally important genes in preimplantation embryos is an accepted marker for unraveling the influence of single factors in studies that are mostly related to artificial reproduction techniques. Such studies, however, often reveal high levels of heterogeneity between single embryos, independently of the influence of factors of interest. A possible explanation for this variation could be the large variety of physiological and environmental factors to which early embryos are exposed and their ability to react to them. Here, we investigated several potentially important parameters of development at the same time, in blastocysts of the wild guinea pig (Cavia aperea) generated in vivo after natural mating. The optimal time for flushing fully developed blastocysts was between 123 and 126 hours after mating. The abundance of POU5F1 (P = 0.042), BAX (P sire. In addition, the identity of the father had a significant influence on POU5F1 (P = 0.049) and STAT3 (P < 0.001) mRNA abundances. These data report that the expression of specific genes in early embryos reflects the entire heterogeneity of their surroundings and that it is a plastic reaction toward a multifactorial environment. PMID:27264741

  18. Effects of Jiedu Quyu Ziyin Recipe on the apoptosis and expressions of bcl-2 and bax mRNA of peripheralblood lymphocyte in MRL/lpr mice%解毒祛瘀滋阴药对MRL/lpr小鼠外周血淋巴细胞凋亡及线粒体跨膜电位的影响

    Institute of Scientific and Technical Information of China (English)

    曹灵勇; 谢志军; 王新昌; 温成平; 范永升

    2010-01-01

    Objective To explore the effects of Jiedu Quyu Ziyin Recipe (JQZR) on the apoptosis and expressions of bcl-2 and bax mRNA of peripheral-blood lymphocyte in MRL/lpr mice. Methods 80 MRL/lpr mice were randomly divided into model group,TCM group, Western medicine group and TCM and Western medicine group,20 mice in each group, meanwhile,20 Kunming mice were selected as normal group, then intragastrically administered normal sodium, JQZR apozem, prednisone suspension and JQZR apozem and prednisone suspension, 0. 5ml every time,once daily for 12 weeks respectively. At the end of the 12th week, peripheral-blood lymphocytes of every mice purified by gradient centrifugation were cultivated for 48 hours ,then the apoptosis was detected by flow cytometry. Furthermore,the expressions of bcl-2 and bax mRNA of peripheral-blood lymphocyte were detected by RT-PCR. Results At Oh or 48h,the apoptosis ratios of PBLC in normal group, TCM group,Western medicine group and TCM and Western medicine group are higher than model group and the differences are significant(P 0. 05) ,even if which are significant between TCM group and TCM and Western medicine group or between model group and TCM and Western medicine group( P 0.05),但中药组和模型组与中西药组比较,差异有显著性(P0.05);西药组与中西药组之间差异无显著性(P>0.05).结论 解毒祛瘀滋阴药能增加MRL/lpr小鼠的PBLC凋亡率并下调其线粒体跨膜电位水平.

  19. Effect of fluorescent whitening agent on the transcription of cell damage-related genes in zebrafish embryos.

    Science.gov (United States)

    Jung, Hyun; Seok, Seung-Hyeok; Han, Ju-Hee; Abdelkader, Tamer Said; Kim, Tae-Hyoun; Chang, Seo-Na; Ko, Ae-Sun; Choi, Seung-Kyu; Lee, Cho-Rong; Seo, Ji-Eun; Byun, Soo-Hyun; Kim, Jung-A; Park, Jae-Hak

    2012-09-01

    7-Diethylamino-4-methylcoumarin (DEMC) is a fluorescent whitening agent (FWAs). There have been some studies on DEMC's protective effects against biological activity but there are few papers about the in vivo toxicity of DEMC. In this study, we used wild-type zebrafish embryos 3 days post fertilization (dpf). Test solutions with DEMC concentrations were negative control (without vehicle), 0 (with vehicle, 0.01% v/v ethanol), 0.25, 0.5, 0.75, 1.0, 1.25, 1.5 and 2 ppm. Embryos and larvae were counted for survival rate and hatching rate. Heart rates were also counted at 2.5 and 3.0 dpf. At 3.0 dpf, quantitative RT-PCR was performed with some samples (0, 0.25, 0.75 and 1.25 ppm) to determine the toxic effect to DEMC by detecting the expression levels of toxic-responsive genes. We used 11 genes, which included oxidative stress-related genes [sod(Mn), sod(Cu,Zn) and hsp70], mitochondrial metabolism-related genes (coxI, pyc, cyt and cyclinG1) and apoptosis-related genes (c-jun, bcl2, bax and p53). High-concentration DEMC-treated groups showed significant different survival rate, hatching rate and heart rate compared with low-concentration DEMC-treated groups. The LC50 of this chemical, 0.959 ppm, was calculated. We also confirmed that some genes in the DEMC exposure groups showed significantly up-regulations in expression levels compared with control groups. We concluded that the fluorescence agent, DEMC, has possible developmental toxicities and alteration effect of gene expression, which are related to oxidative stress, mitochondrial metabolism and apoptosis in zebrafish embryos.

  20. 新生鼠坏死性小肠结肠炎肠上皮细胞Bax表达与细胞凋亡的初步研究%Expression of Bax and apoptosis in enterocytes of neonatal rats with necrotizing enterocolitis

    Institute of Scientific and Technical Information of China (English)

    张勇; 蒋迎佳; 廖志; 叶飘; 郝静梅; 余绍兰; 胡语航

    2014-01-01

    目的 探讨新生鼠坏死性小肠结肠炎(necrotizing enterocolitis,NEC)时肠上皮细胞Bax表达与凋亡相关性.方法 出生当日48只清洁级SD新生鼠采用奇、偶数方法随机分为对照组和NEC模型组,每组24只.对照组由母鼠喂养.模型组与母鼠分开,通过代乳品人工喂养+缺氧-复氧-冷刺激建立新生鼠NEC动物模型.分别于实验开始后24h、48 h、72 h留取大鼠回盲部近段回肠组织行免疫组织化学法检查肠上皮细胞Bax表达,原位末端标记法检测肠上皮细胞凋亡率.结果 与对照组[(666.55±15.77)IOD; (4.73±0.04)%]相比,NEC模型组新生鼠肠上皮细胞Bax表达和细胞凋亡率在实验开始后24 h开始升高[(1 005.06±11.96) IOD; (15.04 ±0.24)%](P<0.01),72 h后达到高峰[(3 340.66±68.72) IOD; (35.65±0.61)%](P<0.01).NEC模型组新生鼠肠上皮细胞凋亡率与肠上皮细胞Bax表达量呈正相关(r=0.94,P<0.01).结论 新生鼠NEC肠上皮细胞凋亡与Bax表达有明确相关性.新生鼠NEC肠上皮细胞可能通过高表达Bax导致肠上皮细胞凋亡.%Objective To investigate the correlation of Bax and apoptosis in enterocytes of neonatal rats with necrotizing enterocolitis (NEC).Methods Forty-eight neonatal rats (1 day) were randomly divided into control group (n =24) and NEC model group (n =24) by use of odd and even.The rats in control group were maternal breast-fed.The rats in NEC model group were separated from their mothers.To be given formula feeding,cold exposure after hypoxic-reoxygenation treatment.The intestinal tissue located at the boundary of ileum and caecum of two groups were gained on the 24 h,48 h and 72 h with which that all rats were sacrificed by cutting neck.Section of intestinal tissue were stained with immunohistochemistry to detect the expression of Bax and were stained with TdT mediated dUTP nick end labeling(TUNEL) to evaluate the apoptosis in each group.Results The integrate optical density (IOD) value of

  1. Effect of gemcitabine heat perfusion chemotherapy combined with carboplatin chemotherapy embolization on serum indexes in patients with hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Wei Zhou; Xing-Yuan Wang; Kun Zhou

    2015-01-01

    Objective:To study the effects of Gemcitabine heat perfusion chemotherapy combined with carboplatin chemotherapy embolization on serum indexes in patients with hepatocellular carcinoma.Methods:90 cases of hepatocellular carcinoma patients were enrolled and randomly divided into two groups. Observation group received gemcitabine heat perfusion chemotherapy combined with carboplatin chemotherapy embolization, control group received gemcitabine conventional perfusion chemotherapy combined with carboplatin chemotherapy embolization. Malignant biological indicators of serum and liver tissue apoptosis regulation of gene expression of the two groups were compared.Results: (1) Serum malignant biological indicators: serum DKK1, TK1, HIF-1 alpha mRNA and protein content of the observation group were lower than that of the control group; (2) Promoting apoptosis gene: MTS1 in liver tissue, Caspase 3 and Bax mRNA and protein contents of the observation group was higher than that of the control group; (3) Apoptosis suppressor genes: liver cancer tissues Plk1, Bcl - 2 and Survivn mRNA and protein contents of the observation group was higher than that of the control group.Conclusion:Gemcitabine hot perfusion chemotherapy plus carboplatin chemotherapy embolism helps to inhibit tumor biological behavior, induce liver cancer cells apoptosis, and it is an ideal treatment for primary liver cancer.

  2. Cytotoxicity, genotoxicity and mechanism of action (vi