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  1. Survivin 2α: a novel Survivin splice variant expressed in human malignancies

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    Honsey Laura E

    2005-03-01

    Full Text Available Abstract Background Survivin and its alternative splice forms are involved in critical cellular processes, including cell division and programmed cell death. Survivin is expressed in the majority of human cancers, but minimally in differentiated normal tissues. Expression levels correlate with tumor aggressiveness and resistance to therapy. Results In the present study, we identify and characterize a novel survivin isoform that we designate survivin 2α. Structurally, the transcript consists of 2 exons: exon 1 and exon 2, as well as a 3' 197 bp region of intron 2. Acquisition of a new in-frame stop codon within intron 2 results in an open reading frame of 225 nucleotides, predicting a truncated 74 amino acid protein. Survivin 2α is expressed at high levels in several malignant cell lines and primary tumors. Functional assays show that survivin 2α attenuates the anti-apoptotic activity of survivin. Subcellular localization and immunoprecipitation of survivin 2α suggests a physical interaction with survivin. Conclusion We characterized a novel survivin splice variant that we designated survivin 2α. We hypothesize that survivin 2α can alter the anti-apoptotic functions of survivin in malignant cells. Thus survivin 2α may be useful as a therapeutic tool in sensitizing chemoresistant tumor cells to chemotherapy.

  2. Immunohistochemical expression of the oncogenic molecules active Stat3 and survivin in benign and malignant salivary gland tumors

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    Nikitakis, Nikolaos G.; Scheper, Mark A.; Papanicolaou, Vasileios S.; Sklavounou, Alexandra; Sauk, John J.

    2009-01-01

    Objective Signal transducer and activator of transcription 3 (Stat3) and survivin have been shown to exert oncogenic effects in various human neoplasms. The purpose of this study was to evaluate the expression of the tyrosine phosphorylated (active) Stat3 and survivin in various benign and malignant salivary gland tumors (SGTs). Study design Eighty-six SGTs (65 malignant and 21 benign tumors of various histopathologic subtypes) were immunohistochemically stained with anti-survivin or anti-phosphorylated tyrosine-705 (p-tyr) Stat3 antibodies. Immunohistochemical reactivity was graded in a semi-quantitative manner; a combined score of immunohistochemical positivity (0–6) was calculated for each tumor by adding the individual scores for percentage of tumor cells (0–3) and intensity of staining (0–3). Results Survivin was immunohistochemically detected in all studied benign and malignant SGTs; p-tyr Stat3 was also detected in the majority (91%) of SGTs. The average combined scores for survivin and p-tyr Stat3 immunohistochemical expression in the studied malignant SGTs was 4.40 and 3.35, respectively; the corresponding combined scores for survivin and p-tyr Stat3 in the studied benign SGTs were 4.37 and 3.22, respectively. No statistically significant differences (p>0.05) in p-tyr Stat3 or survivin expression were detected between the benign and malignant groups, or among the various examined histopathological subtypes of SGTs. In contrast, normal salivary gland elements in the vicinity of the studied tumors revealed only weak and focal survivin or p-tyr Stat3 immunoreactivity, mainly localized to ductal and mucous cells. Conclusions Our data indicate an almost universal expression of activated Stat3 and survivin in benign and malignant SGTs. Considering the well-established proliferative and anti-apoptotic properties of these molecules and their functional interrelationship, selective targeting techniques against Stat3 and/or survivin may represent promising

  3. Inhibition of fibroblast growth factor 2-induced apoptosis involves survivin expression, protein kinase Cα activation and subcellular translocation of Smac in human small cell lung cancer cells

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    Desheng Xiao; Kuansong Wang; Jianhua Zhou; Huiqiu Cao; Zhenghao Deng; Yongbin Hu; Xiahui Qu; Jifang Wen

    2008-01-01

    To investigate the mechanism by which fibroblast growth factor 2 (FGF-2) inhibits apoptosis in the human small cell lung cancer cell line H446 subjected to serum starvation,apoptosis was evaluated by flow cytometry, Hoechst 33258 staining, caspase-3 activity, and DNA fragmentation.Survivin expression induced by FGF-2 and protein kinase Cα (PKCα) translocation was detected by subcellular fractionation and Western blot analysis. In addition, FGF-2-induced release of Smac from mitochondria to the cytoplasm was analyzed by Western blotting and immunofluorescence.FGF-2 reduced apoptosis induced by serum starvation and up-regulated survivin expression in H446 cells in a dosedependent and time-dependent manner, and inhibited caspase-3 activity. FGF-2 also inhibited the release of Smac from mitochondria to the cytoplasm induced by serum starvation and increased PKCα translocation from the cytoplasm to the cell membrane. In addition, PKC inhibitor inhibited the expression of survivin. FGF-2 up-regulates the expression of survivin protein in H446 cells and blocks the release of Smac from mitochondria to the cytoplasm. PKCα regulated FGF-2-induced survivin expression. Thus, survivin, Smac,and PKCα might play important roles in the inhibition of apoptosis by FGF-2 in human small cell lung cancer cells.

  4. Survivin expression and its clinical significance in pancreatic cancer

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    Lee Kyung Shik

    2005-10-01

    Full Text Available Abstract Background Survivin, an inhibitor of apoptosis is expressed in several human cancers. Its expression is known to be associated with poor clinical outcome, but not widely studied in pancreatic cancer. We performed this study to determine the survivin expression in pancreatic cancer and its clinical significance as a prognostic factor. Methods We performed immunohistochemical staining for survivin, p53, and Bax in formalin-fixed, paraffin-embedded block from forty-nine pancreatic tissues. To determine the association with clinical course, we reviewed the patients' clinical record. Results Of the 49 cases of pancreatic cancer, 46 cases (93.9% were positive for survivin expression. There was no significant association between survivin expression and p53 or bax. For clinicopathological parameters, perineural invasion was more common in survivin positive and venous invasion was more common in survivin negative (p = 0.041 and 0.040, respectively. Responsiveness to chemotherapy appeared to be slightly better in patients with low survivin expression. Conclusion Survivin expression may be associated with venous or perineural invasion, indicating metastatic route, and seems to have a potential as a predictive marker for chemotherapy. Further study of large scale is required to determine the clinical significance of survivin expression in pancreatic cancer.

  5. EFFECT OF SURVIVIN-siRNA-MEDIATED GENE SILENCING ON SURVIVIN EXPRESSION IN OSTEOSARCOMA CELL LINE MG-63

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    WANG Jing-wei; CAO Lei; WANG Yu; FU Jun; TIAN Hai-mei; LIU Yi; ZHANG Wei

    2006-01-01

    Objective: To study the inhibition of expression of Survivin gene by synthesized short Survivin-siRNA in osteosarcoma cell line MG-63. Methods: Chemically synthesized short Surviving-siRNA was transfected into osteosarcoma cell line MG-63. The Survivin mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM). The biological morphology and growth inhibition of MG-63 were observed with light microscopy and MTT assay, respectively. Results: Short siRNA targeting Survivin down-regulated the transcription and the protein level of Survivin oncogene. The proliferation of osteosarcoma cell line MG-63 was inhibited after transfection. Conclusion: Chemically synthesized short Survivin-siRNA can effectively inhibit Survivin expression and cell proliferation inosteosarcoma cell line MG-63 Survivin-siRNA mediated Survivin gene silencing may be a useful therapeutic strategy for osteosarcoma.

  6. Delivery of a survivin promoter-driven antisense survivin-expressing plasmid DNA as a cancer therapeutic: a proof-of-concept study

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    Lin KY

    2016-05-01

    Full Text Available Kun-Yuan Lin,1 Siao Muk Cheng,2 Shing-Ling Tsai,2 Ju-Ya Tsai,1 Chun-Hui Lin,1 Chun Hei Antonio Cheung1,2 1Department of Pharmacology, College of Medicine, National Cheng Kung University, Tainan, Taiwan, ROC; 2Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan, ROC Abstract: Survivin is a member of the inhibitor-of-apoptosis proteins family. It is overexpressed in many different cancer types but not in the differentiated normal tissue. In addition, overexpression of survivin promotes cancer cell survival and induces chemotherapeutic drug resistance, making it an attractive target for new anticancer interventions. Despite survivin being a promising molecular target for anticancer treatment, it is widely accepted that survivin is only a “semi-druggable” target. Therefore, it is important to develop a new strategy to target survivin for anticancer treatment. In this study, we constructed a novel survivin promoter-driven full-length antisense survivin (pSur/AS-Sur expression plasmid DNA. Promoter activity assay revealed that the activity of the survivin promoter of pSur/AS-Sur correlated with the endogenous expression of survivin at the transcriptional level in the transfected A549, MDA-MB-231, and PANC-1 cancer cells. Western blot analysis showed that liposomal delivery of pSur/AS-Sur successfully downregulated the expression of survivin in A549, MBA-MB-231, and PANC-1 cells in vitro. In addition, delivery of pSur/AS-Sur induced autophagy, caspase-dependent apoptosis, and caspase-independent apoptosis as indicated by the increased LC3B-II conversion, autophagosome formation, caspase-9/-3 and poly(ADP-ribose polymerase-1 cleavage, and apoptosis-inducing factor nuclear translocation in A549, MBA-MB-231, and PANC-1 cells. Importantly, liposomal delivery of pSur/AS-Sur was also capable of decreasing the proliferation of the survivin/MDR1 coexpressing multidrug-resistant KB-TAX50 cancer cells and

  7. The Survivin −31 Snp in Human Colorectal Cancer Correlates with Survivin Splice Variant Expression and Improved Overall Survival

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    Anna G. Antonacopoulou

    2010-01-01

    Full Text Available Background: Survivin is involved in the regulation of cell division and survival, two key processes in cancer. The majority of studies on survivin in colorectal cancer (CRC have focused on protein expression and less is known about the expression of survivin splicing variants or survivin gene polymorphisms in CRC. In the present study, the mRNA levels of the five known isoforms of survivin as well as survivin protein were assessed in matched normal and neoplastic colorectal tissue. Moreover, the 9386C/T and −31G/C polymorphisms were investigated.

  8. The expression and clinical significance of survivin gene in leukemia

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    王艳

    2006-01-01

    Objective To investigate the expression of survivin in leukemia and the prognostic significance in acute leukemia(AL). Methods The expression of survivin mRNA was measured in 105 AL and 21 chronic myelogenous leukemia (CML) patients with semi-quantity reverse transcription (RT)-PCR.15 adults were tested as normal

  9. Expression of Survivin in pancreatic cancer and its correlation to expression of Bcl-2

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    Jian-Guo Qiao; Yu-Qing Zhang; Yu-Chun Yin; Zui Tan

    2004-01-01

    AIM: To investigate the expression of Survivin in pancreatic cancer and its correlation to the expression of Bcl-2.METHODS: Survivin and Bcl-2 expressions were examined by immunohistochemistry in 42 tissue samples from pancreatic cancer and 10 from normal pancrease. RESULTS: No survivin expression was detected in the tissue samples from normal pancrease, while it was detected in 34 of 42 tissue samples from pancreatic cancer (81.95%).There was a correlation between survivin expression and differentiation and stages of pancreatic cancer. Survivin positive cases were strongly correlated to Bcl-2 expression (28/30 vs 6/12, P<0.05).CONCLUSION: Overexpression of survivin plays an important role in the development and progression of pancreatic cancer, and correlates to the expression of Bcl-2. Survivin expression can be used as a prognostic factor.

  10. Expressions of MGMT and Survivin in Colorectal Carcinoma

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    QI Xiao-li; WANG Fa-liang; BO Ai-hua; HOU Jin-chao; NIU Shu-lei

    2008-01-01

    Objective:To investigate the expressions of O6-methyguanine-DNA methytransferase(MGMT) and Survivin in colorectal carcinoma and their clinical significance.Methods:Formalin-fixed,paraffin-embedded specimens from polypus and colorectal carcinoma were examined with streptavidin-biotin peroxidase(S-P)immunohistochemical technique for the expressions of MGMT and Survivin.Results:We found that there were significant differences in MGMT and Survivin between polypus and colorectal carcinoma.Expression of MGMT was correlated with ages and lymph node metastasis while Survivin was associated with lymph node metastasis only.Meanwhile,the expression of MGMT was correlated with Survivin (P<0.01,r=0.65).But there was no significant difference between male and female and the different depth of infiltration.Conclusion:It is concluded that the abnormal expressions of MGMT and Survivin were associated with the degree of malignancy of colorectal tumor.They possibly could be useful indexes for the primary screening and prognosis of colorectal carcinoma.ExaminatiOn of them may have an important guiding significance in the chemotherapy strategy.

  11. Counteraction of the Antiapoptotic Protein Survivin by Diverting Expression to its Proapoptotic Splice Variant Survivin-2B

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    2010-01-01

    negatively regulated by low glucose. [17] Further, glucose restriction activates the longevity- associated histone and protein deacetylase, SIRT1 ...pattern of decreasing with low glucose. We also studied SIRT1 because it was already reported to epigenetically silence survivin transcription and...furthermore to be upregulated during glucose restriction. Indeed, SIRT1 increased with glucose restriction, in opposite manner as survivin and survivin-2B

  12. PROGNOSTIC SIGNIFICANCE OF EXPRESSION OF SURVIVIN IN ACUTE LEUKEMIA

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    王晓娟; 戴国仪; 曹利民; 王国华; 朱慧芬; 张悦; 沈关心

    2002-01-01

    Objective: To investigate the expression of survivin gene and its significance in acute leukemia. Methods: The expression of surviving in 134 acute leukemia patients and 4 leukemia cell lines was detected by RT-PCR and immunofluorescence analysis. Results: We detected survivin expression in 78 of 134 acute leukemia patients and all the cell lines but not in normal controls and anemia patients. Survivin gene expression correlated with a lower white blood cell count, which was 11×109/L and 48×109/L in the positive and negative group respectively (P<0.01 by the Mann-Whitney test). In 55 cases of FAB M1/M2/M3, it was associated with leukemic cell maturation(P<0.01 by the Fisher test). Survivin expression was strongly related to survival time of acute leukemia patients (P<0.05). Conclusion: These data suggest that survivin expression may be considered as a new unfavorable prognostic factor for acute leukemia due to its important role in apoptosis inhibition that influences disease outcome.

  13. Expression of Survivin Gene and Its Relationship with Clinical Multidrug Resistance in Osteosarcoma

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    NIETao; DAIMin; ZONGShizhang

    2005-01-01

    Objective: To study the expression of survivin and its relationship with clinical multidrug resistance in osteosarcoma. Methods: By using immunohistochemistry (S-P) method, the expression of Survivin in osteosarcoma, osteochondroma and normal osseous tissue, and the expression of P-glycoprotein in osteosarcoma was detected. Results: Survivin positive expression rate was 65.71% in osteosarcoma, but no expression of Survivin was detectable in osteochondroma and normal osseous tissue. The positive expression rate of Survivin was significantly associated with Enneking clinical stages and histological typing (WHO), but no relationship was found among Survivin expression and age, sex and tumor location. The positive expression rate of P-glycoprotein was 45.71%. There was a significant correlation between Survivin and p-glycoprotein. Conclusion: Survivin overexpression was significantly associated with clinical multidrug resistance in osteosarcoma. It could be a potential target for treatment of osteosarcoma.

  14. Survivin promotes the invasion of human colon carcinoma cells by regulating the expression of MMP‑7.

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    Gao, Fei; Zhang, Yuqin; Yang, Feng; Wang, Peng; Wang, Wenjun; Su, Yan; Luo, Weiren

    2014-03-01

    Increased expression levels of survivin are crucial for invasion activity in several types of human cancer, including colon carcinoma. However, the molecular mechanisms whereby survivin regulates cancer invasion have not been completely elucidated. To the best of our knowledge, this study is the first to investigate the role of matrix metalloprotease‑7 (MMP‑7) in cell invasion that is induced by survivin by using in vitro assays, including western blot, immunofluorescence and qPCR analyses. The results demonstrated that the ectopic expression of survivin significantly promoted the invasive activity of colon carcinoma cells (SW620 and HCT‑116) and resulted in increased levels of MMP‑7 activation. By contrast, the small interfering RNA (siRNA)‑based knockdown of survivin markedly reduced cell migration and led to a dose‑dependent decrease in MMP‑7 expression levels. Compared with the controls, knockdown of MMP‑7 by siRNA in colon carcinoma cells led to reduced invasion ability, whereas no obvious changes were observed when MMP‑7 expression was silenced in survivin‑overexpressing colon carcinoma cells. These findings demonstrate that MMP‑7 is crucial for survivin‑mediated invasiveness, suggesting that the survivin‑mediated MMP‑7 signaling pathway is a potential therapeutic target for the treatment of colon carcinoma.

  15. A PRELIMINARY STUDY ON SURVIVIN AND BCL-2 EXPRESSION IN CERVICAL CARCINOMAS

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    2001-01-01

    Objective To study the expression of a novel inhibitor of apptosis and survivin in cervical carcinoma and its relationship to the expression of Bcl-2.Methods Using SP immunohistochemical technique, we examined the expression of survivin and Bcl-2 in 59 cervical invasive squamous cell carcinomas.Results Survivin was expressed in 41 of 59 cases(69.5%) of cervical carcinomas. In contrast, no expression of survivin in normal cervical tissues was observed. Overexpression of survivin was related to the tumor grade and clinical stage. Survivin positive cases were strongly associated with Bcl-2 expression(80% versus 35.7%;P<0.005).Conclusion Apoptosis inhibition by survivin abnormal expression, alone or in cooperation with Bcl-2, may participate in the onset and progression of cervical carcinoma. Survivin is a new diagnostic/therapeutic target in cervical cancer.

  16. Transcriptional inhibition of p21{sup WAF1/CIP1} gene (CDKN1) expression by survivin is at least partially p53-dependent: Evidence for survivin acting as a transcription factor or co-factor

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    Tang, Lei [Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, NY (United States); Pre-Doctoral Chinese Fellowship Student, Second West China Hospital, Sichuan University, Sichuan (China); Ling, Xiang; Liu, Wensheng; Das, Gokul M. [Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, NY (United States); Li, Fengzhi, E-mail: fengzhi.li@roswellpark.org [Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, NY (United States)

    2012-05-04

    Highlights: Black-Right-Pointing-Pointer Survivin inhibits the expression of p21 protein, mRNA and promoter activity. Black-Right-Pointing-Pointer Survivin neutralizes p53-induced p21 expression and promoter activity. Black-Right-Pointing-Pointer Survivin physically interacts with p53 in cancer cells. Black-Right-Pointing-Pointer Genetic silencing of endogenous survivin upregulates p21 in p53 wild type cancer cells. Black-Right-Pointing-Pointer Both p53 and survivin interacts on the two p53-binding sites in the p21 promoter. -- Abstract: Growing evidence suggests a role for the antiapoptotic protein survivin in promotion of cancer cell G1/S transition and proliferation. However, the underlying mechanism is unclear. Further, although upregulation of p21{sup WAF1/CIP1} by p53 plays an important role in p53-mediated cell G1 arrests in response to various distresses, it is unknown whether survivin plays a role in the regulation of p21{sup WAF1/CIP1} expression. Here, we report that exogenous expression of survivin in p53-wild type MCF-7 breast cancer cells inhibits the expression of p21{sup WAF1/CIP1} protein, mRNA and promoter activity, while the survivin C84A mutant and antisense failed to do so. Cotransfection experiments in the p53 mutant H1650 lung cancer cell line showed that survivin neutralizes p53-induced p21{sup WAF1/CIP1} expression and promoter activity. Importantly, genetically silencing of endogenous survivin using lentiviral survivin shRNA also enhances endogenous p21 in p53 wild type cancer cells, suggesting the physiological relevance of the fining. We further demonstrated that both p53 and survivin interacts on the two p53-binding sites in the p21{sup WAF1/CIP1} promoter (-2313 to -2212; -1452 to -1310), and survivin physically interacts with p53 in cancer cells. Together, we propose that survivin may act as a transcription factor or cofactor to interact with p53 on the p21{sup WAF1/CIP1} promoter leading to the inhibition of p21{sup WAF1/CIP1

  17. Expression of survivin and p53 modulates honokiol-induced apoptosis in colorectal cancer cells.

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    Lai, Ying-Jiun; Lin, Chien-I; Wang, Chia-Lin; Chao, Jui-I

    2014-11-01

    Honokiol is a small biphenolic compound, which exerts antitumor activities; however, the precise mechanism of honokiol-induced apoptosis in the human colorectal cancer cells remains unclear. Here, we show that survivin and p53 display the opposite role on the regulation of honokiol-induced apoptosis in the human colorectal cancer cells. Honokiol induced the cell death and apoptosis in various colorectal cancer cell lines. Moreover, honokiol elicited the extrinsic death receptor pathway of DR5 and caspase 8 and the intrinsic pathway of caspase 9. The common intrinsic and extrinsic downstream targets of activated caspase 3 and PARP protein cleavage were induced by honokiol. Interestingly, honokiol reduced anti-apoptotic survivin protein and gene expression. Transfection with a green fluorescent protein (GFP)-survivin-expressed vector increased the colorectal cancer cell viability and resisted the honokiol-induced apoptosis. Meantime, honokiol increased total p53 and the phosphorylated p53 proteins at Ser15 and Ser46. The p53-wild type colorectal cancer cells were exhibited greater cytotoxicity, apoptosis and survivin reduction than the p53-null cancer cells after treatment with honokiol. Together, these findings demonstrate that the existence of survivin and p53 can modulate the honokiol-induced apoptosis in the human colorectal cancer cells.

  18. RELATIONSHIP BETWEEN THE EXPRESSIONS OF SURVIVIN AND THE PROGNOSTIC RELATED FACTORS IN BREAST CANCER

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    SHEN Jing-hua; WANG Xiao-juan; SU He-ba-te; ZHAO Xiao-xia; TAO Ge-si

    2005-01-01

    Objective: To study the relationship between the Survivin expression and the histological grade, status of ER,expression of PS2 and the prognosis of patients with primary invasive breast cancer. Method: By using LSAB and SP immunohistochemical method, the expression of Survivin, PS2 and ER in 95 cases of invasive breast cancer were detected.Results: the positive rate of Survivin was 70.5% (67/95) and the expression of Survivin was positively related to the histological grade and status of PS2 and ER. The survival time after operation of patients without expression of Survivin was longer than those with positive Survivin. Conclusion: These data suggest that Survivin expression may be considered as a new unfavorable prognostic factor of breast cancer.

  19. Vitamin D Receptor, Retinoid X Receptor, Ki-67, Survivin, and Ezrin Expression in Canine Osteosarcoma

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    John Davies

    2012-01-01

    Full Text Available Canine osteosarcoma (OS is an aggressive malignant bone tumor. Prognosis is primarily determined by clinical parameters. Vitamin D has been postulated as a novel therapeutic option for many malignancies. Upon activation, vitamin D receptors (VDRs combine with retinoid receptor (RXR forming a heterodimer initiating a cascade of events. Vitamin D's antineoplastic activity and its mechanism of action in OS remain to be clearly established. Expression of VDR, RXR, Ki-67, survivin, and ezrin was studied in 33 archived, canine OS specimens. VDR, RXR, survivin, and ezrin were expressed in the majority of cases. There was no statistically significant difference in VDR expression in relationship with tumor grade, type, or locations or animal breed, age, and/or sex. No significant association (p=0.316 between tumor grade and Ki-67 expression was found; in particular, no difference in Ki-67 expression between grades 2 and 3 OSs was found, while a negative correlation was noted between Ki-67 and VDR expression (ρ=−0.466, a positive correlation between survivin and RXR expression was found (p=0.374. A significant relationship exists between VDR and RXR expression in OSs and proliferative/apoptosis markers. These results establish a foundation for elucidating mechanisms by which vitamin D induces antineoplastic activity in OS.

  20. Relationship between expression of Smac and Survivin and apoptosis of primary hepatocellular carcinoma

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    Shi-Ting Bao; Shui-Qing Gui; Mu-Sheng Lin

    2006-01-01

    BACKGROUND: The second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein with low pI (Smac/DIABLO) was recently identiifed as a protein that is released from mitochondria in response to apoptotic stimuli and promotes apoptosis by antagonizing inhibitor of apoptosis proteins. Furthermore, Smac/DIABLO plays an important regulatory role in the sensitization of cancer cells to both immune-and drug-induced apoptosis. However, little is known about the clinical signiifcance of Smac/DIABLO in various cancers including hepatocellular carcinoma (HCC). This study was undertaken to investigate the expression of Smac and Survivin and their relationship with the apoptosis in primary HCC. METHODS:The expression of Smac and Survivin proteins was evaluated by immunohistochemistry. The mRNA expression of Smac and Survivin was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) in HCC tissues of 50 patients, para-carcinoma tissues of 20 patients, and normal liver tissues of 15 patients. RESULTS: Smac mRNA was detected by RT-PCR in HCC tissues of 21 (42.0%) of the 50 patients, para-carcinoma tissues of 19 (95.0%) of the 20 patients, and normal liver tissues of 15 (100%) of the 15 patients. Survivin mRNA was found in HCC tissues of 46 of the 50 patients, para-carcinoma tissues of 2 of the 20 patients, and normal liver tissues of 0 of 15 patients. Immunohistochemistry revealed Smac protein in HCC tissues of 20 patients (40.0%), in para-carcinoma tissues of 18 patients (90.0%), and normal liver tissues of 15 patients (100.0%). The expression of Smac was signiifcantly different in HCC tissues and non-HCC tissues. Survivin protein was found in HCC tissues in 45 patients, para-carcinoma tissues in 2 patients, and normal liver tissues in none of the patients. The expression of Survivin was signiifcantly different in HCC tissues and non-HCC tissues. CONCLUSION: Smac inhibits apoptosis of HCC cells by suppression of Survivin, and the

  1. Increased expression of survivin in gastric cancer patients and in first degree relatives

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    J. Yu; Leung, W K; Ebert, M P A; Ng, E K W; Go, M Y Y; Wang, H.B.; Chung, S C S; Malfertheiner, P; Sung, J. J. Y.

    2002-01-01

    Survivin was recently described as an apoptosis inhibitor. Its pathogenic role in gastric cancer is largely unknown. Expression of survivin in gastric cancer and non-cancer first-degree relatives, and its association with apoptosis and cyclo-oxygenase-2 expression was investigated. Fifty gastric cancer, 30 non-cancer first-degree relatives, 20 normal controls and five gastric cancer cell lines were studied. Survivin and cyclo-oxygenase-2 were evaluated by reverse transcriptase-polymerase chai...

  2. Survivin expression in canine epidermis and in canine and human cutaneous squamous cell carcinomas.

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    Bongiovanni, Laura; Colombi, Isabella; Fortunato, Carmine; Della Salda, Leonardo

    2009-10-01

    Survivin, a member of the inhibitor of apoptosis protein (IAP) family, is ubiquitously expressed during tissue development, undetectable in most normal tissues, but re-expressed in most cancers, including skin malignancies. Expression of survivin was evaluated retrospectively in 19 canine cutaneous squamous cell carcinomas (SCCs; one in situ; 16 well differentiated; one invasive, one lymph node metastasis) and 19 well differentiated SCCs from human beings. Seven specimens of normal canine skin were included. Immunohistochemical expression of full-length survivin was determined using a commercially available antibody. In addition, apoptotic rate [Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labelling index (TUNEL) index] and mitotic index (MI), counting mitoses in 10 high power fields (HPF), were determined. Scattered survivin positive nuclei were identified in the epidermal basal cell layer of normal canine skin. Nuclear survivin expression was identified in 18 of 19 human and in all canine SCCs, mainly along the base of the tumour cell population. Cytoplasmic survivin expression was rarely observed in human SCCs and in 84.2% of canine SCCs. The TUNEL index ranged from 0.1 to 2.6 in human beings and from 7.5 to 69.4 in dogs, while MIs ranged from 0 to 4 in human beings and dogs. No correlation was found between survivin expression and apoptotic or mitotic rates. Canine and human tumours showed similar nuclear survivin expression, indicating similar functions of the molecule. We demonstrated survivin expression in normal adult canine epidermis. Increased nuclear survivin expression in pre-neoplastic and neoplastic lesions demonstrates a possible association of survivin with development of SCCs in human beings and dogs.

  3. Expression of Survivin, CDK4, Ki-67 and Clinical Significance in Pediatric Acute Leukemia

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    ZHANG Liuqing; LIU Jing; LIN Hanhua; HU Qun; LIU Aiguo; HU Ying

    2006-01-01

    The expression of Survivin, CDK4 and Ki-67 and the clinical significance in pediatric acute leukemia (AL) were investigated. The expression of Survivin, CDK4 and Ki-67 proteins was detected by using immunohistochemical assay in 37 children with AL and 10 children with normal bone marrow as controls. The positive expression rate of Survivin, CDK4 and Ki-67 was 45.9 %,56.8 %, and 40.5 % respectively in 37 AL children, which was significantly higher than in control group accordingly (P<0.05). The expression of Survivin was positively correlated with CDK4(P=0.007) and Ki-67 (P=0.008). In conclusion, all Survivin, CDK4 and Ki-67 proteins are over-expressed in pediatric AL and involved in the modulation of apoptosis and proliferation in pediatric AL.

  4. Carbenoxolone Induces Apoptosis and Inhibits Survivin and Survivin-ΔEx3 Genes Expression in Human Leukemia K562 Cells

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    Z. Sanaat

    2011-12-01

    Full Text Available Background and the purpose of the study: Leukemia is a malignant disorder of the blood progenitor/stem cells which is characterized by abnormal proliferation of white blood cells. Although anti-cancer drugs induce apoptosis in cancerous cells, drug resistance is the significant problem mainly due to over-expression of inhibitors of apoptosis proteins (IAPs such as survivin. In this content, it has been reported that an anti-inflammatory drug, Carbenoxolone (CBX, could induce apoptosis and growth inhibition in several types of cancerous cells. In the present study, effects of CBX on apoptosis and level of the expression of survivin gene and its ΔEx3 splicing variant have were evaluated in K562 cells.Methods: K562 cells were cultured and treated with different concentrations of CBX (50-300 μM at different time intervals (12-48 hrs. Trypan blue exclusion test was used to evaluate cell viability. Fluorescent microscopy (Acridine Orange/Ethidium Bromide double staining and DNA fragmentation assay were used to study apoptosis. The expression level of survivin and its ΔEx3 splice variant were studied by RT- PCR.Results and Major Conclusion: It was found that both growth inhibition and apoptosis occurred in K562 cells. In addition, down-regulation of survivin and survin-ΔEx3 were observed, after 2-4 hrs treatment with 150 μM of CBX. However, the expression level of survivin and its ΔEx3 splice variant increased in subsequent time (6-12 hrs nearly to the level of control cells. From the results of this study, it may be concluded that CBX can be considered as a candidate for further studies in CML treatment, especially in the case of drug- resistant leukemia cells.

  5. SURVIVIN AND TUMOR

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    宋文哲; 宋燕; 叶剑桥; 邱东涛

    2003-01-01

    As a new member of IAP (inhibitors of apoptosis protein) family, survivin has potent anti-apoptotic activities, and involves in the mitosis and angiogenesis. Researches have demonstrated that surviving is a tumor-specific anti-apoptotic factor, expressed in fetal tissues, and common human cancers, while not in normal, terminally differentiated adult tissues. The overexpression of survivin in tumor tissues is correlated with poor prognosis of the patients. Survivin can be used as a prognostic factor and a new target in tumor targeting therapy.

  6. Expression of PLK1 and survivin in non-Hodgkin's lymphoma treated with CHOP

    Institute of Scientific and Technical Information of China (English)

    Lin LIU; Min ZHANG; Ping Z0U

    2008-01-01

    Aim:The present study was designed to investigate the expression of Polo-like kinase 1 (PLK1) and survivin in non-Hodgkin's lymphoma (NHL).Methods:The expression of PLKI and survivin were detected with immunohistochemical techniques.Results:The expression rate of PLKi and survivin were 63.6% (56/ 88) and 79.5% (70/88) in NHL,respectively.PLKI expression correlated with systemic symptoms,lactate dehydrogenase levels,and international prognostic index scores in B-NHL and T-NHL,while survivin did not.Conclusion:PLK 1 and survivin are both overexpressed in NHL.There is a significant relationship be-tween the overexpression of PLK1 and clinical features.

  7. Increased spontaneous apoptosis, but not survivin expression, is associated with histomorphologic response to neoadjuvant chemoradiation in rectal cancer.

    LENUS (Irish Health Repository)

    McDowell, Dermot T

    2009-11-01

    Survivin has been shown to be an important mediator of cellular radioresistance in vitro. This study aims to compare survivin expression and apoptosis to histomorphologic responses to neoadjuvant radiochemotherapy (RCT) in rectal cancer.

  8. TFF3 and survivin expressions associate with a lower survival rate in gastric cancer.

    Science.gov (United States)

    Meng, Jia-Rong; Tang, Hui-Zhong; Zhou, Kai-Zong; Shen, Wu-Hong; Guo, He-Yi

    2013-11-01

    Trefoil factor 3 (TFF3) and survivin with functions of inhibiting apoptosis are involved in the gastric cancer by overexpression. The purpose of this study is to examine the expression of TFF3 and survivin in patients' tissue samples with gastric cancer and analyze the relationship between the protein expression and the different clinical records. By studying the expressions of TFF3 and survivin in gastric cancer through immunohistochemical staining and examining the survival rate via Kaplan-Meier analysis for gastric cancer patients, we found that the TFF3 and survivin positive expressions have a significant relationship with the lower survival rate comparing to that of negative expressions in the analyzed patients (P TFF3 and survivin expressions have the lowest survival rate. TFF3 or survivin positive expression correlates with the lymph node metastasis, metastasis, and TNM stages of gastric cancer. Survival analysis indicates that survival rate has a close relationship with the age, tumor histology, tumor differentiation, degree of infiltration, lymph node metastasis, distant metastasis, and TNM stages (P TFF3 and survivin expressions play a vital role in gastric cancer development, and these two proteins are important markers for prognosis in gastric cancer. Patients with gastric cancer can increase the survival rate through an earlier diagnosis and appropriate treatment.

  9. Caspase-3 and survivin expression in pediatric neuroblastoma and their roles in apoptosis

    Institute of Scientific and Technical Information of China (English)

    王家祥; 郑树

    2004-01-01

    Background Neuroblastoma, one of the common tumors in children, possesses the feature of natural regression that might be related to apoptosis caspase-3 and survivin are believed to respectively induce and inhibit apoptosis. We investigated the expression of caspase-3 and survivin in pediatric neuroblastoma and the role that these genes played in apoptosis.Methods The expression of caspase-3 and survivin in pediatric neuroblastoma tissue samples was detected using in situ hybridization, ter mintuesal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and immunohistochemical staining. The role that these genes played in apoptosis was then evaluated.Results A converse correlation was observed between the expression of survivin and caspase-3. When survivin was expressed at high levels in neuroblastoma samples, caspase-3 expression was downregulated, and the apoptotic index decreased simultaneously.Conclusion There is a converse correlation between the expression of caspase-3 and the expression of survivin in neuroblastoma cells, indicating that caspase-3 might induce apoptosis, and survivin may inhibit this process.

  10. Expression of Survivin, p53 and its relationship with apoptosis, proliferation in hepatocellular carcinoma(HCC)

    Institute of Scientific and Technical Information of China (English)

    Wentao Hui; Ying Zan; Xijng Wang; Huafeng kang; Haitao Guan; Xiaobin Ma

    2008-01-01

    Objective:To investigate the expression of Survivinp53 and its relationship with apoptosis, proliferation in hepatocellular carcinoma (HCC).Methods:The expression of Survivin, p53 and the proliferation of tumor cells marked by proliferation cell nuclear antigen (PCNA) in 42 cases of HCC were assessed by immunohistochemical method.TUNEL was used to detect apoptosis.Results:Survivin protein was expressed in 30 of 42 cases of HCC(71.4%) and in 4 of 34 cases of adjacent cirrhosis tissues(11.8%).Expression of Survivin protein was negative in 10 cases of normal tissues.Survivin protein positive expression rate in HCC was significantly higher than adjacent cirrhosis tissues and normal tissues(P 0.05).Conclusion:There is a marked increased expression of Survivin in HCC, which may play an important role in breaking the balance of proliferation and apoptosis of HCC cells.The correlation between Survivin and p53 expression in HCC indicates that cooperation between Survivin and p53 plays a certain role in occurrence and/or development of HCC.

  11. Expression of survivin mRNA in peritoneal lavage fluid from patients with gastric carcinoma

    Institute of Scientific and Technical Information of China (English)

    王振宁; 徐惠绵; 姜莉; 周欣; 鲁翀; 张学

    2004-01-01

    Background Peritoneal dissemination is the most common pattern of metastasis in advanced gastric carcinoma with serosal invasion. In the present study, we reported the clinical relevance of a new diagnostic method involving RT-PCR, using survivin as the target gene, for the detection of free cancer cells in peritoneal washes.Methods Intraoperative peritoneal washes were obtained from 48 patients who underwent surgery for gastric cancer. RT-PCR analysis with primers specific for survivin and conventional cytological examinations were both performed.Results Survivin mRNA was not detected in any peritoneal wash samples from patients with benign disease, but was detected in 28 of 48 samples taken from patients with gastric cancer and in all metastastic nodules. Survivin expression in the peritoneal cavity significantly correlated with depth of cancer invasion, lymph node metastasis, and TNM stage. There were 92% of clinically evident peritoneal metastasis cases showed detectable survivin expression. The combination of survivin RT-PCR and cytological examination yielded positive results in 66.7% (32/48) of patients with gastric cancer, much higher than the results produced by cytological method alone. Conclusions Survivin mRNA detected in peritoneal lavage fluid might indicate the presence of free cancer cells in the peritoneal cavity. The high sensitivity of the RT-PCR-based survivin assay suggests that survivin serves as a molecular marker for detecting peritoneal micrometastasis. Its ubiquitous expression in peritoneal cancer cells and metastatic nodules also suggests a promising future therapeutic strategy based on survivin inhibition for cases of gastric cancer involving peritoneal metastasis.

  12. Differential localization and high expression of SURVIVIN splice variants in human embryonic stem cells but not in differentiated cells implicate a role for SURVIVIN in pluripotency

    Directory of Open Access Journals (Sweden)

    Amber N. Mull

    2014-03-01

    Full Text Available The BIRC5 gene encodes the oncofetal protein SURVIVIN, as well as four additional splice variants (ΔEx3, 2B, 3B and 2α. SURVIVIN, an inhibitor of apoptosis, is also a chromosomal passenger protein (CPP. Previous results have demonstrated that SURVIVIN is expressed at high levels in embryonic stem cells and inhibition of SURVIVIN function results in apoptosis, however these studies have not investigated the other four splice variants. In this study, we demonstrate that all variants are expressed at significantly higher levels in human embryonic stem (hES cells than in differentiated cells. We examined the subcellular localization of the three most highly expressed variants. SURVIVIN displayed canonical CPP localization in mitotic cells and cytoplasmic localization in interphase cells. In contrast, SURVIVIN–ΔEx3 and SURVIVIN–2B did not localize as a CPP; SURVIVIN–ΔEx3 was found constitutively in the nucleus while SURVIVIN–2B was distributed along the chromosomes during mitosis and also to the mitotic spindle poles. We used inducible shRNA against SURVIVIN to inhibit expression in a titratable fashion. Using this system, we reduced the mRNA levels of these three variants to approx. 40%, resulting in a concomitant reduction of OCT4 and NANOG mRNA, suggesting a role for the SURVIVIN variants in pluripotency.

  13. Expression of Yes-associated protein modulates Survivin expression in primary liver malignancies.

    Science.gov (United States)

    Bai, Haibo; Gayyed, Mariana F; Lam-Himlin, Dora M; Klein, Alison P; Nayar, Suresh K; Xu, Yang; Khan, Mehtab; Argani, Pedram; Pan, Duojia; Anders, Robert A

    2012-09-01

    Hepatocellular carcinoma and intrahepatic cholangiocarcinoma account for 95% of primary liver cancer. For each of these malignancies, the outcome is dismal; incidence is rapidly increasing, and mechanistic understanding is limited. We observed abnormal proliferation of both biliary epithelium and hepatocytes in mice after genetic manipulation of Yes-associated protein, a transcription coactivator. Here, we comprehensively documented Yes-associated protein expression in the human liver and primary liver cancers. We showed that nuclear Yes-associated protein expression is significantly increased in human intrahepatic cholangiocarcinoma and hepatocellular carcinoma. We found that increased Yes-associated protein levels in hepatocellular carcinoma are due to multiple mechanisms including gene amplification and transcriptional and posttranscriptional regulation. Survivin, a member of the inhibitors-of-apoptosis protein family, has been reported as an independent prognostic factor for poor survival in both hepatocellular carcinoma and intrahepatic cholangiocarcinoma. We found that nuclear Yes-associated protein expression correlates significantly with nuclear Survivin expression for both intrahepatic cholangiocarcinoma and hepatocellular carcinoma. Furthermore, using mice engineered to conditionally overexpress Yes-associated protein in the liver, we found that Survivin messenger RNA expression depends upon Yes-associated protein levels. Our findings suggested that Yes-associated protein contributes to primary liver tumorigenesis and likely mediates its oncogenic effects through modulating Survivin expression.

  14. Expression of survivin in Human Non-Hodgkin Lymphoma and Its Correlation with Proliferation and Angiogenesis

    Institute of Scientific and Technical Information of China (English)

    LI Jiansha; WU Huanming

    2006-01-01

    In order to investigate the expression change of survivin in non-Hodgkin lymphoma (NHL) and its possible effects on NHL development, the expression of survivin, Ki-67, caspase3 and FⅧRAg in reactive lymphoid hyperplasia (RH) and NHL was detected by immunohistochemical assay, and apoptosis index (AI) in RH and NHL by TUNEL analysis. The results showed that the expression of survivin is significantly higher in aggressive NHL than in indolent NHL (P<0.01), while there was no statistically significant difference between RH and indolent NHL (P>0.05). The expression of survivin had a significantly positive correlation with the expression of Ki-67 and FⅧRAg (r=0.6495, 0.6635, respectively, both P<0.01), and a negative correlation with the expression of caspase3 and AI (r=-0.5820, -0.6013, respectively, P<0.01). It was suggested that survivin may contribute to the progression of NHL by playing an important role in promoting cell proliferation, inhibiting cell apoptosis and enlisting angiogenesis. Survivin expression is closely related to malignant grade and therefore may be considered an important prognostic factor of NHL.

  15. Effects of AFP gene silencing on Survivin mRNA expression inhibition in HepG2 cells.

    Science.gov (United States)

    Fang, Z L; Fang, N; Han, X N; Huang, G; Fu, X J; Xie, G S; Wang, N R; Xiong, J P

    2015-04-10

    We investigated the effects of alpha-fetoprotein (AFP) gene silencing on Survivin expression in HepG2 cells. Small interfering RNA technology was used to downregulate AFP expression in HepG2 cells. An enzyme-linked immunosorbent assay was used to measure AFP concentration in the supernatant before and after transfection. An MTT assay was used to detect cell proliferation activity before and after transfection. We performed flow cytometric analysis to detect the cell apoptosis rate, and reverse transcription-polymerase chain reaction to detect Survivin mRNA levels before and after transfection. Forty-eight hours after transfection, AFP concentration in the supernatant of the experimental group significantly decreased, hepatocellular carcinoma cell growth was inhibited by 43.1%, and the apoptosis rate increased by 24.3%. Survivin mRNA expression was reduced by 78.0% in HepG2 cells. These indicators in the control group and in the blank group did not change significantly. Silencing of AFP expression in HepG2 cells can effectively inhibit the growth of hepatoma cells and promote apoptosis, which may be useful for reducing intracellular Survivin mRNA levels.

  16. Expression of survivin, a novel apoptosis inhibitor and cell cycle regulatory protein, in human gliomas

    Institute of Scientific and Technical Information of China (English)

    焦保华; 姚志刚; 耿少梅; 左书浩

    2004-01-01

    @@ Recently, a novel anti-apoptosis gene, named survivin,was identified as a structurally unique member of the inhibitor of apoptosis protein (lAP) family. The gene is located on chromosome 17q25. Survivin is a 16.5 kDa protein that is expressed in vivo in common human cancers, but not in normal adjacent tissue,1 during the G2/M phase of the cell cycle. Survivin expression is turned off during fetal development and not found in nonneoplastic adult human tissue, and it is turned on in most common human cancers. We investigated the expression of survivin in 50 patients with human gliomas, and determined its association with cell apoptosis and cell proliferation, and its impact on tumor progression and prognosis.

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

    Institute of Scientific and Technical Information of China (English)

    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.

  18. C-reactive protein inhibits survivin expression via Akt/mTOR pathway downregulation by PTEN expression in cardiac myocytes.

    Directory of Open Access Journals (Sweden)

    Beom Seob Lee

    Full Text Available C-reactive protein (CRP is one of the most important biomarkers for arteriosclerosis and cardiovascular disease. Recent studies have shown that CRP affects cell cycle and inflammatory process in cardiac myocytes. Survivin is also involved in cardiac myocytes replication and apoptosis. Reduction of survivin expression is associated with less favorable cardiac remodeling in animal models. However, the effect of CRP on survivin expression and its cellular mechanism has not yet been studied. We demonstrated that treatment of CRP resulted in a significant decrease of survivin protein expression in a concentration-dependent manner in cardiac myocytes. The upstream signaling proteins of survivin, such as Akt, mTOR and p70S6K, were also downregulated by CRP treatment. In addition, CRP increased the protein and mRNA levels of PTEN. The siRNA transfection or specific inhibitor treatment for PTEN restored the CRP-induced downregulation of Akt/mTOR/p70S6K pathway and survivin protein expression. Moreover, pretreatment with a specific p53 inhibitor decreased the CRP-induced PTEN expression. ERK-specific inhibitor also blocked the p53 phosphorylation and PTEN expression induced by CRP. Our study provides a novel insight into CRP-induced downregulation of survivin protein expression in cardiac myocytes through mechanisms that involved in downregulation of Akt/mTOR/p70S6K pathway by expression of PTEN.

  19. EXPRESSION AND SIGNIFICANCE OF SURVIVIN mRNA IN LUNG CANCER TISSUE MICROARRAY DETECTED BY FISH

    Institute of Scientific and Technical Information of China (English)

    Xin-yun Wang; Xing-ye Wu; Zhi Yao; Yan Li; Ting Liu; Hai-yan Zheng; Cong-zhong Zhu; Cui-yun Sun; Ai-xiang Wang; Min Zhao

    2005-01-01

    Objective To investigate the expression of Survivin mRNA in lung cancer tissue microarray (TMA) by fluorescence in situ hybridization (FISH) method, and determine the role and significance of it in lung cancer genesis and progress. Methods The expression of Survivin mRNA was detected by FISH method and TMA technology. Fifty-four cases of lung cancer and 10 cases of normal lung tissue were examined. Results Survivin mRNA was expressed in 66.7% (36/54) of lung cancer; the positive ratio of lung cancer was significantly higher than that of normal lung tissue (0/10; x2= 15.238, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in poor differentiated cancer (20/24, 83.3%) than moderate and well differentiated cancer (16/30, 53.3%; x2= 5.40, P <0.05). The positive ratio of Survivin mRNA was significantly higher in group with lymph node metastasis (27/32, 84.4%) than without lymph node metastasis (9/22, 40.9%; x2= 11.084, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in stage Ⅲ-Ⅳ(12/13, 92.3%) than stage Ⅰ - Ⅱ (24/41, 58.5%; x2= 5.066, P < 0.05). Conclusion Survivin mRNA highly expresses in lung cancer, which is related to the progress and malignant behavior. Survivin may play a promoting role in lung cancer genesis and progress and provide a basis for estimating prognosis and treatment.

  20. Tunicamycin promotes apoptosis in leukemia cells through ROS generation and downregulation of survivin expression.

    Science.gov (United States)

    Lim, Eun Jin; Heo, Jeonghoon; Kim, Young-Ho

    2015-08-01

    Tunicamycin (TN), one of the endoplasmic reticulum stress inducers, has been reported to inhibit tumor cell growth and exhibit anticarcinogenic activity. However, the mechanism by which TN initiates apoptosis remains poorly understood. In the present study, we investigated the effect of TN on the apoptotic pathway in U937 cells. We show that TN induces apoptosis in association with caspase-3 activation, generation of reactive oxygen species (ROS), and downregulation of survivin expression. P38 MAPK (mitogen-activated protein kinase) and the generation of ROS signaling pathway play crucial roles in TN-induced apoptosis in U937 cells. We hypothesized that TN-induced activation of p38 MAPK signaling pathway is responsible for cell death. To test this hypothesis, we selectively inhibited MAPK during treatment with TN. Our data demonstrated that inhibitor of p38 (SB), but not ERK (PD) or JNK (SP), partially maintained apoptosis during treatment with TN. Pre-treatment with NAC and GSH markedly prevented cell death, suggesting a role for ROS in this process. Ectopic expression of survivin in U937 cells attenuated TN-induced apoptosis by suppression of caspase-3 cleavage, mitochondrial membrane potential, and cytochrome c release in U937 cells. Taken together, our results show that TN modulates multiple components of the apoptotic response of human leukemia cells and raise the possibility of a novel therapeutic strategy for hematological malignancies.

  1. Survivin expression and prognostic significance in pediatric malignant peripheral nerve sheath tumors (MPNST.

    Directory of Open Access Journals (Sweden)

    Rita Alaggio

    Full Text Available Malignant peripheral nerve sheath tumors (MPNST are very aggressive malignancies comprising approximately 5-10% of all soft tissue sarcomas. In this study, we focused on pediatric MPNST arising in the first 2 decades of life, as they represent one the most frequent non-rhabdomyosarcomatous soft tissue sarcomas in children. In MPNST, several genetic alterations affect the chromosomal region 17q encompassing the BIRC5/SURVIVIN gene. As cancer-specific expression of survivin has been found to be an effective marker for cancer detection and outcome prediction, we analyzed survivin expression in 35 tumor samples derived from young patients affected by sporadic and neurofibromatosis type 1-associated MPNST. Survivin mRNA and protein expression were assessed by Real-Time PCR and immunohistochemical staining, respectively, while gene amplification was analyzed by FISH. Data were correlated with the clinicopathological characteristics of patients. Survivin mRNA was overexpressed in pediatric MPNST and associated to a copy number gain of BIRC5; furthermore, increased levels of transcripts correlated with a higher FNCLCC tumor grade (grade 1 and 2 vs. 3, p = 0.0067, and with a lower survival probability (Log-rank test, p = 0.0038. Overall, these data support the concept that survivin can be regarded as a useful prognostic marker for pediatric MPNST and a promising target for therapeutic interventions.

  2. Survivin expression and prognostic significance in pediatric malignant peripheral nerve sheath tumors (MPNST).

    Science.gov (United States)

    Alaggio, Rita; Turrini, Riccardo; Boldrin, Daniela; Merlo, Anna; Gambini, Claudio; Ferrari, Andrea; Dall'igna, Patrizia; Coffin, Cheryl M; Martines, Annalisa; Bonaldi, Laura; De Salvo, Gian Luca; Zanovello, Paola; Rosato, Antonio

    2013-01-01

    Malignant peripheral nerve sheath tumors (MPNST) are very aggressive malignancies comprising approximately 5-10% of all soft tissue sarcomas. In this study, we focused on pediatric MPNST arising in the first 2 decades of life, as they represent one the most frequent non-rhabdomyosarcomatous soft tissue sarcomas in children. In MPNST, several genetic alterations affect the chromosomal region 17q encompassing the BIRC5/SURVIVIN gene. As cancer-specific expression of survivin has been found to be an effective marker for cancer detection and outcome prediction, we analyzed survivin expression in 35 tumor samples derived from young patients affected by sporadic and neurofibromatosis type 1-associated MPNST. Survivin mRNA and protein expression were assessed by Real-Time PCR and immunohistochemical staining, respectively, while gene amplification was analyzed by FISH. Data were correlated with the clinicopathological characteristics of patients. Survivin mRNA was overexpressed in pediatric MPNST and associated to a copy number gain of BIRC5; furthermore, increased levels of transcripts correlated with a higher FNCLCC tumor grade (grade 1 and 2 vs. 3, p = 0.0067), and with a lower survival probability (Log-rank test, p = 0.0038). Overall, these data support the concept that survivin can be regarded as a useful prognostic marker for pediatric MPNST and a promising target for therapeutic interventions.

  3. Targeting Hsp90 with small molecule inhibitors induces the over-expression of the anti-apoptotic molecule, survivin, in human A549, HONE-1 and HT-29 cancer cells

    Directory of Open Access Journals (Sweden)

    Lyu Kevin W

    2010-04-01

    Full Text Available Abstract Background Survivin is a dual functioning protein. It inhibits the apoptosis of cancer cells by inhibiting caspases, and also promotes cancer cell growth by stabilizing microtubules during mitosis. Since the molecular chaperone Hsp90 binds and stabilizes survivin, it is widely believed that down-regulation of survivin is one of the important therapeutic functions of Hsp90 inhibitors such as the phase III clinically trialed compound 17-AAG. However, Hsp90 interferes with a number of molecules that up-regulate the intracellular level of survivin, raising the question that clinical use of Hsp90 inhibitors may indirectly induce survivin expression and subsequently enhance cancer anti-drug responses. The purpose of this study is to determine whether targeting Hsp90 can alter survivin expression differently in different cancer cell lines and to explore possible mechanisms that cause the alteration in survivin expression. Results Here, we demonstrated that Hsp90 inhibitors, geldanamycin and 17-AAG, induced the over-expression of survivin in three different human cancer cell lines as shown by Western blotting. Increased survivin mRNA transcripts were observed in 17-AAG and geldanamycin-treated HT-29 and HONE-1 cancer cells. Interestingly, real-time PCR and translation inhibition studies revealed that survivin was over-expressed partially through the up-regulation of protein translation instead of gene transcription in A549 cancer cells. In addition, 17-AAG-treated A549, HONE-1 and HT-29 cells showed reduced proteasomal activity while inhibition of 26S proteasome activity further increased the amount of survivin protein in cells. At the functional level, down-regulation of survivin by siRNA further increased the drug sensitivity to 17-AAG in the tested cancer cell lines. Conclusions We showed for the first time that down-regulation of survivin is not a definite therapeutic function of Hsp90 inhibitors. Instead, targeting Hsp90 with small

  4. Construction and Expression of Human Survivin and Preparation of Its Polyclonal Antibody

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hai-hong; ZHANG Xi-zhen; ZHAO Dong-hai; SHI He-liang; YU Yong-hui; WU Yong-ge; YU Xiang-hui; KONG Wei

    2008-01-01

    Survivin,a novel member of inhibitor ofapoptosis(IAP) protein family,is aberrantly expressed in cancer but undetectable in normal,differentiated adult tissues.The cancer-specific expression of survivin,coupled with its importance in inhibiting cell death and in regulating cell division makes it a useful diagnostic marker of cancer and a potential target for cancer treatment.Survivin cDNA amplified from the total RNA of 293 cells through RT-PCR was cloned into prokaryotic expression vector pRSET-B.The recombinant plasmid pRSET-B-Surv was expressed in E.coli BL21,and the relative molecule mass(Mr) of expressed fusion protein was approximately 21000.The recombinantprotein was purified through Ni2~ affinity chromatography column and characterized by SDS-PAGE and Western blot.The purified recombinant protein was then injected into rabbits,and antisurvivin polyclonal antibody with a high titer was obtained.

  5. Survivin is a guardian of the intestinal stem cell niche and its expression is regulated by TGF-β.

    Science.gov (United States)

    Martini, Eva; Schneider, Evelyn; Neufert, Clemens; Neurath, Markus F; Becker, Christoph

    2016-11-01

    As an inhibitor of apoptosis (IAP) family member, Survivin is known for its role during regulation of apoptosis. More recently its function as a cell cycle regulator has become evident. Survivin was shown to play a pivotal role during embryonic development and is highly expressed in regenerative tissue as well as in many cancer types. We examined the function of Survivin during mouse intestinal organogenesis and in gut pathophysiology. We found high expression of Survivin in experimentally induced colon cancer in mice but also in colon tumors of humans. Moreover, Survivin was regulated by TGF-β and was found to be highly expressed during mucosal healing following intestinal inflammation. We identified that expression of Survivin is essential early on in life, as specific deletion of Survivin in Villin expressing cells led to embryonic death around day 12 post coitum. Together with our recent study on the role of Survivin in the gut of adult mice our data demonstrate that Survivin is an essential guardian of embryonic gut development and adult gut homeostasis protecting the epithelium from cell death promoting the proliferation of intestinal stem and progenitor cells.

  6. EXPRESSION OF SURVIVIN AND E-CADHERIN IN BREAST CANCER

    Institute of Scientific and Technical Information of China (English)

    TIAN Xiao-feng; LIU Ji-hong; WANG Li-fen; FENG XIAO-Mei; YAO Ji-hong

    2005-01-01

    Objective: Survivin is a member of the inhibitor of apoptosis (IAP) family, and is involved in the regulation of cell division. E-cadherin functionally belongs to transmembrane glycoproteins family, it is responsible for intercellular junction mechanism that is crucial for the mutual association of vertebrate cells. These genes are thought to be associated with cancer aggression. This study was to investigate the relationship between surviving gene, E-cadherin expression and invasion clinicopathological features of breast cancer. Methods: The expression of surviving gene and E-cadherin were detected by SP immunohistochemical technique in tissues of 66 breast cancer, 20 breast fibroadenoma and 20 adjacent breast tissue. Results: The positive rate of surviving gene expression in breast cancer was 42.2%, significantly higher (P=0.025) than those in breast fibroadenoma (35.0%), and adjacent breast tissue (10.0%). The positive rate of E-cadherin in the groups of adjacent breast tissue, breast fibroadenoma and breast cancer were 100%, 100% and 42.4%, there was significant difference between the group of benign and malignant tumor (P=0.005). The positive rate of surviving in breast cancer with local lymph node metastasis was significant higher than that in breast cancer without lymph node metastasis (P=0.01), and E-cadherin in breast cancer with local lymph node metastasis was significant lower than that without lymph node metastasis (P=o.o1). There was no significant difference among the groups of pathological types and TNM stages in the expression of surviving (P=0.966 & P=0.856), but there was significant difference in the expression of E-cadherin among these groups (P=0.01 & P=0.023). Conclusion: The loss or decrease of E-cadherin expression may promote the exfoliation of cancerous cells from original tissues, and surviving gene may promote the viability of the exfoliated cancer cells and the formation of new metastasis focus. These 2 factors cooperate with each other

  7. Expression of the Apoptosis Inhibitor Survivin and its correlation with Thymidine Kinase and Axillary Lymph Node Metastasis in Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    1 Introduction Many molecular factors have been demonstrated to interfere with cellular proliferation and programmed cell death.One of these factors is a recently discovered member of the “inhibitor of apoptosis protein(IAP)” called survivin. Survivin is abundantly expressed in most solid and hematologic malignancies, but undetectable in normal adult tissues. Interference with survivin function induces pleiotropic cell-division defects and apoptosis. Cytosolic thymidine kinase (TK) is a marker for prolifera...

  8. Arsenic Trioxide Inhibits Proliferation in K562 Cells by Changing Cell Cycle and Survivin Expression

    Institute of Scientific and Technical Information of China (English)

    伍晓菲; 陈智超; 刘仲萍; 周浩; 游泳; 黎纬明; 邹萍

    2004-01-01

    To study the mechanisms involved in the inhibition of chronic myeloid leukemic cells (K562) proliferation induced by arsenic trioxide (As2O3) and to explore the potential role of Survivin, an inhibitor of apoptosis protein, in the regulation of As2O3 induced cell apoptosis, K562 cells were cultured with As2O3 of different concentrations. Cells were collected for proliferation analysis by MTT assay. Cell cycle distribution and cell apoptosis were analyzed by flow cytometry.Expression of Survivin protein and mRNA were detected by flow cytometry and RT-PCR, respectively. Our results showed that As2O3 (2-10 μmol/L) inhibited K562 cells growth effectively, but it did not induce cells apoptosis significantly. The percentage of K562 cells at G2/M phase increased in proportion to As2O3 concentrations, and the expression of Survivin mRNA and content of Survivin protein was up-regulated accordingly. It is concluded that As2 O3 inhibited K562 cells growth by inducing cell cycle arrest mainly at G2/M phase. Over-expression of Survivin gene and protein might be one of the possible mechanisms contributing to K562 cells' resistance to As2O3-induced apoptosis.

  9. Changes of Survivin mRNA and Protein Expression during Paclitaxel Treatment in Breast Cancer Cells

    Institute of Scientific and Technical Information of China (English)

    XIONG Huihua; YU Shiying; ZHUANG Liang; XIONG Hua

    2007-01-01

    In order to investigate the role of antiapoptosis gene, survivin in the resistance to palcitaxel, the expression of survivin mRNA and protein in the process of paclitaxel treatment in breast cancer cell line MCF-7 was detected. MCF-7 cells were incubated with paclitaxel at different concentrations. The growth inhibition rate of MCF-7 was investigated by tetrazolium bromide (MTT) colorimetry. The change of apoptosis was detected by Annexin-V/PI methods. The changes in the expression of survivin mRNA and protein were studied by reverse transcription polymerase chain reaction (RT-PCR) and Western-blot assay respectively. The growth inhibition rate of MCF-7 was increased in a concentration- and time-dependent manner. Paclitaxel of higher concentration could effectively induce apoptosis in MCF-7 cells after 48 h, while the expression of survivin was increased at early time (within 6 h) and decreased after 24 h regardless of treatment concentrations of paclitaxel. It suggested that tumor cells might evade the paclitaxel-induced cell cycle arrest and apoptosis by increasing the level of survivin at early treatment time.

  10. 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.

  11. Immunohistochemical study of p16 INK4A and survivin expressions in cervical squamous neoplasm

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    Tan Geok

    2010-01-01

    Full Text Available Introduction:Cervical cancer is the second most common cancer affecting Malaysian women. Despite the implementation of pap smear screening, many women are still diagnosed only in the advanced stage of cervical cancer. This could partly be due to failure of detection of its precursor lesions; hence the need to search for novel biomarkers to assist in the screening and diagnosis of cervical neoplasia. This study aims to determine the expression of p16INK4A and survivin as possible predictive biomarkers in cervical squamous neoplasm. Material and Methods: This is a retrospective study on 201 cases of cervical neoplasm comprising of 129 cervical intraepithelial neoplasia (CIN and 72 squamous cell carcinoma (SCC. All samples were evaluated by two independent observers using p16INK4A and survivin monoclonal antibodies. The p16 INK4A expression was graded as negative, focal and diffuse positivity. The intensity for survivin expression was graded as weak, moderate and intense. Results: It is seen that p16 INK4A expression in CIN 1, CIN 2 and CIN 3 were 25.4%, 42.9% and 95.9% respectively. Majority of SCC (98.6% showed p16 INK4A expression. Survivin expressions in CIN 1, CIN 2, CIN 3 and SCC were 56.7%, 33.4%, 87.5% and 98.6%. There was a linear relationship between increasing grade of CIN and p16 INK4A expressions. Conclusion: Our study showed that p16 INK4A expressions correlate well with the increasing grade of CIN. Although survivin does not correlate well to the increasing grade of CIN, it could be useful in differentiating CIN 3 from SCC.

  12. Evaluation of tissue and urinary survivin expression in non-muscle-invasive bladder cancer

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    S. Sharaf

    2012-12-01

    Conclusion: Urinary survivin is a useful marker for non-invasive detection of non-muscle-invasive bladder cancer recurrence. Its detection is better using ELISA technique than WB and there is no correlation between its expression in tissue and urine.

  13. Testicular expression of survivin and human telomerase reverse transcriptase (hTERT) associated with spermatogenic function in infertile patients

    Institute of Scientific and Technical Information of China (English)

    Steffen Weikert; Frank Christoph; Wolfgang Schulze; Hans Krause; Carsten Kempkensteffen; Martin Schostak; Kurt Miller; Mark Schrader

    2006-01-01

    Aim: To characterize the coexpression of survivin, an inhibitor of apoptosis (IAF), and human telomerase reverse transcriptase (hTERT) in human testes with varying spermatogenic function. Methods: Transcript levels of survivin mRNA and hTERT mRNA were determined in normal testes (n = 11) and testes with defective spermatogenesis (n = 28) using real-time reverse-transcription polymerase chain reaction (RT-PCR). The histological work-up was performed according to a modified Johnsen score. Results: Expressions of both survivin and hTERT were highest at median levels of 96.8 and 709 in normal spermatogenesis and dropped to 53.3 and 534 in testes with postmeiotic spermatogenic arrest (n = 10). In severe spermatogenic failure (n = 18), survivin expression was lacking in most specimens (n = 16), whereas at least low levels of testicular hTERT expression were largely detectable with a normalized expression of 73 in premeiotic spermatogenic arrest (n = 7) and 45 in patients with Sertoli cell-only syndrome (SCOS)(n = 3). Both survivin and hTERT expressions increased with a progressing Johnsen score (P for trend = 0.001).Conclusion: Although both survivin and hTERT are correlated with spermatogenic function, they show different expression patterns in testes of infertile patients. These findings substantiate results from studies in the rodent testis suggesting a predominant expression of survivin in meiotically dividing germ cells.

  14. High density lipoprotein (HDL)-associated sphingosine 1-phosphate (S1P) inhibits macrophage apoptosis by stimulating STAT3 activity and survivin expression

    DEFF Research Database (Denmark)

    Feuerborn, Renata; Becker, Susen; Potì, Francesco;

    2016-01-01

    reticulum-dependent apoptosis was induced by exposure of macrophages to etoposide or thapsigargin/fukoidan, respectively. RESULTS: Cell death induced by these compounds was inhibited by S1P as inferred from reduced annexin V binding, TUNEL staining, and caspase 3, 9 and 12 activities. S1P induced expression...

  15. Establishment of a Tumor-bearing Mouse Model Stably Expressing Human Tumor Antigens Survivin and MUC1 VNTRs

    Institute of Scientific and Technical Information of China (English)

    ZHANG Li-xing; DU Jian-shi; WANG Yu-qian; LIU Chen-lu; XIA Qiu; ZHANG Xi-zhen; CONG Xian-ling; ZHANG Hai-hong

    2012-01-01

    The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1(MUC1)(VNTRs),namely,VR1012-S and VR1012-VNTR(VNTR=variable number of tandem repeat),were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line(B16)stably expressing survivin and MUC1 VNTRs(MS+B16)was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS+B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs(MS)expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS+B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×105 MS+B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline(PBS),a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs.

  16. Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721

    Institute of Scientific and Technical Information of China (English)

    Sheng-Quan Cheng; Wen-Liang Wang; Wei Yan; Qing-Long Li; Li Wang; Wen-Yong Wang

    2005-01-01

    AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells.METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SW), were constructed by ligating into the vector,pSupperssorNeo (pSuNeo) digested with restriction enzymes Xba I and Sa/I and the designed double-chain RNAi primers. A cell model of SMMC-7721 after treatment with RNAi was prepared by transfecting SMMC-7721 cells with the lipofectin transfection method. Strept-avidinbiotin-complex (SABC) immunohistochemical staining and RT-PCR were used to detect survivin gene expressions in SMMC-7721 cells. Flow cytometry was used for the cell cycle analysis. Transmission electron microscopy was performed to determine whether RNAi induced cell apoptosis, and the method of measuring the cell growth curve was utilized to study the growth of SMMC-7721 cells before and after treatment with RNAi.RESULTS: The eukaryotic expression vector of survivin gene RNAi and pSuNeo-SW, were constructed successfully. The expression level of survivin gene in SMMC-7721 cells was observed. After the treatment of RNAi, the expression of survivin gene in SMMC-7721 cells was almost absent,apoptosis index was increased by 15.6%, and the number of cells was decreased in G2/M phase and the cell growth was inhibited.CONCLUSION: RNAi can exert a knockdown of survivin gene expression in SMMC-7721 cells, and induce apoptosis and inhibit the growth of carcinoma cells.

  17. SF002-96-1, a new drimane sesquiterpene lactone from an Aspergillus species, inhibits survivin expression

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    Silke Felix

    2013-12-01

    Full Text Available Survivin, a member of the IAP (inhibitor of apoptosis gene family, is overexpressed in virtually all human cancers and is functionally involved in the inhibition of apoptosis, regulation of cell proliferation, metastasis and resistance to therapy. Because of its upregulation in malignancy, survivin has currently attracting considerable interest as a new target for anticancer therapy. In a screening of approximately 200 strains of imperfect fungi for the production of inhibitors of survivin promoter activity, a new drimane sesquiterpene lactone, SF002-96-1, was isolated from fermentations of an Aspergillus species. The compound inhibited survivin promoter activity in transiently transfected Colo 320 cells in a dose dependent manner with IC50 values of 3.42 µM (1.3 µg/mL. Moreover, it also reduced mRNA levels and protein synthesis of survivin and triggered apoptosis.

  18. REAL-TIME DETECTION OF SURVIVIN mRNA EXPRESSION IN CERVICAL CANCER CELL LINES USING MOLECULAR BEACON IMAGING

    Institute of Scientific and Technical Information of China (English)

    An Ruifang; He Dalin; Xue Yan; Wang Shu; Xie Li; Zhao Jun; Wang Xinyang; Yang Lili

    2006-01-01

    Objective To detect the expression of survivin mRNA in cervical cancer cell lines using molecular beacon imaging technology. Methods Human cervical cancer cells (HeLa and SiHa) and human fetal lung fibroblast HFL-I were cultured in vitro. After adding 100 nmol/L survivin mRNA molecular beacon, the fluorescent signals were observed under fluorescent microscope. The expressions of survivin in cervical cancer cells and HFL-I cell were examined by immunocytochemical streptravidin-biothin peroxidase (SP) assay at the same time. Results Two kinds of survivin mRNA molecular beacon, with different color fluorescence, had strong fluorescent signal in cervical cancer cell lines, and the signal in SiHa cell line was stronger, but these signals were not found in HFL-I ; Immunocytochemical staining of positive survivin was located in the cytoplasm of cervical cancer cell lines HeLa and SiHa, whereas, no expression of survivin was detected in HFL-I cell line. Conclusion The technology of molecular beacon imaging can be used to detect the expression of survivin mRNA in viable cells successfully, and may provide a new approach to the diagnosis of early stage cervical cancer and the following-up in the clinic.

  19. Expression of survivin and matrix metalloproteinases in adenocarcinoma and squamous cell carcinoma of the uterine cervix.

    Science.gov (United States)

    Yoshida, Hiroyuki; Sumi, Toshiyuki; Hyun, Yooji; Nakagawa, Eri; Hattori, Kanae; Yasui, Tomoyo; Morimura, Mina; Honda, Ken-Ichi; Nakatani, Tatsuya; Ishiko, Osamu

    2003-01-01

    Cervical cancer can be classified into two histological types: squamous cell carcinoma (SCA) and adenocarcinoma (ACA). Reportedly ACA has poorer prognoses, metastasizes more easily to lymph nodes, and is more resistant to radiotherapy than SCA. To clarify the cause of characteristic differences between these histological types, we examined the expressions of apoptosis inhibiting and tumor-invasion related factors in both histological types. We reviewed the 34 cases of cervical cancer (17 ACA, 17 SCA) that had surgery as their initial treatment at Osaka City University Medical School Hospital between 1996 and 2001. The differences of survivin, and matrix metalloproteinase (MMP-2, and MMP-7) expressions between both histological types were immunohistochemically assayed, and the correlation between the expression of each protein and clinicopathological characteristics was analyzed. Survivin was expressed significantly stronger in ACA cases (p=0.035). The number of patients who expressed MMP-2 and MMP-7 simultaneously was significantly higher in SCA cases (p=0.039). MMP-2 and MMP-7 had tendencies to be expressed stronger in SCA (p=0.057 and p=0.084, respectively). These results suggest that the differences of the expression of survivin (an apoptosis inhibiting factor), MMP-2, and MMP-7 (tumor-invasion related factors) between ACA and SCA were causes of the characteristic differences between the two histological types.

  20. Chemoresistance of CD133(+) colon cancer may be related with increased survivin expression.

    Science.gov (United States)

    Lee, Mi-Ra; Ji, Sun-Young; Mia-Jan, Khalilullah; Cho, Mee-Yon

    2015-07-31

    CD133, putative cancer stem cell marker, deemed to aid chemoresistance. However, this claim has been challenged recently and we previously reported that patients with CD133(+) colon cancer have benefit from 5-fluorouracil (5-FU) chemotherapy incontrast to no benefit in patients with CD133(-) cancer. To elucidate the role of CD133 expression in chemoresistance, we silenced the CD133 expression in a colon cancer cell line and determined its effect on the biological characteristics downstream. We comparatively analyzed the sequential changes of MDR1, ABCG2, AKT1 and survivin expression and the result of proliferation assay (WST-1 assay) with 5-FU treatment in CD133(+) and siRNA-induced CD133(-) cells, derived from Caco-2 colon cancer cell line. 5-FU treatment induced significantly increase of the mRNA expression of MDR1, ABCG2 and AKT1genes, but not protein level. CD133 had little to no effect on the mRNA and protein expression of these genes. However, survivin expression at mRNA and protein level were significantly increased in CD133(+) cells compared with siRNA-induced CD133-cells and Mock (not sorted CD133(+) cells) at 96 h after siRNA transfection. The cytotoxicity assay demonstrated notable increase of chemoresistance to 5-FU treatment (10 μM) in CD133(+) cells at 96 h after siRNA transfection. From this study, we conclude that CD133(+) cells may have chemoresistance to 5-FU through the mechanism which is related with survivin expression, instead of MDR1, ABCG2 and AKT1 expression. Therefore a survivin inhibitor can be a new target for effective treatment of CD133(+) colon cancer.

  1. Expression of the Apoptosis Inhibitor Survivin and its correlation with Thymidine Kinase and Axillary Lymph Node Metastasis in Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    Jian-Ping WU; Yun-Feng ZHOU; Zhi-Guo LUO; Ming-Sheng ZHANG

    2005-01-01

    @@ 1 Introduction Many molecular factors have been demonstrated to interfere with cellular proliferation and programmed cell death. One of these factors is a recently discovered member of the "inhibitor of apoptosis protein(IAP)" called survivin. Survivin is abundantly expressed in most solid and hematologic malignancies, but undetectable in normal adult tissues. Interference with survivin function induces pleiotropic cell-division defects and apoptosis. Cytosolic thymidine kinase (TK) is a marker for proliferating cells and TK is one of several key enzymes involved in DNAmetabolism that phosphorylates thymidine to thymidine mono-phosphate. This study was aimed to detect the expression of suvivin and TK in breast cancer, and to explore a possible relationship between survivin expression and axillary lymph node metastasis.

  2. Chemoresistance of CD133{sup +} colon cancer may be related with increased survivin expression

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Mi-Ra; Ji, Sun-Young; Mia-Jan, Khalilullah [Department of Pathology, Yonsei University, Wonju College of Medicine, Wonju (Korea, Republic of); Cho, Mee-Yon, E-mail: meeyon@yonsei.ac.kr [Department of Pathology, Yonsei University, Wonju College of Medicine, Wonju (Korea, Republic of); Institute of Genomic Cohort, Yonsei University, Wonju College of Medicine, Wonju (Korea, Republic of)

    2015-07-31

    CD133, putative cancer stem cell marker, deemed to aid chemoresistance. However, this claim has been challenged recently and we previously reported that patients with CD133{sup +} colon cancer have benefit from 5-fluorouracil (5-FU) chemotherapy incontrast to no benefit in patients with CD133{sup −} cancer. To elucidate the role of CD133 expression in chemoresistance, we silenced the CD133 expression in a colon cancer cell line and determined its effect on the biological characteristics downstream. We comparatively analyzed the sequential changes of MDR1, ABCG2, AKT1 and survivin expression and the result of proliferation assay (WST-1 assay) with 5-FU treatment in CD133{sup +} and siRNA-induced CD133{sup −} cells, derived from Caco-2 colon cancer cell line. 5-FU treatment induced significantly increase of the mRNA expression of MDR1, ABCG2 and AKT1genes, but not protein level. CD133 had little to no effect on the mRNA and protein expression of these genes. However, survivin expression at mRNA and protein level were significantly increased in CD133{sup +} cells compared with siRNA-induced CD133-cells and Mock (not sorted CD133{sup +} cells) at 96 h after siRNA transfection. The cytotoxicity assay demonstrated notable increase of chemoresistance to 5-FU treatment (10 μM) in CD133{sup +} cells at 96 h after siRNA transfection. From this study, we conclude that CD133{sup +} cells may have chemoresistance to 5-FU through the mechanism which is related with survivin expression, instead of MDR1, ABCG2 and AKT1 expression. Therefore a survivin inhibitor can be a new target for effective treatment of CD133{sup +} colon cancer. - Highlights: • We evaluate the role of CD133 in chemoresistance of colon cancer. • We compared the chemoresistance of CD133{sup +} cells and siRNA-induced CD133{sup −} cells. • CD133 had little to no effect on MDR1, ABCG2 and AKT1 expression. • Survivin expression and chemoresistance were increased in CD133{sup +} colon cancer cells.

  3. Induction of apoptosis and inhibition of proliferation in Hep-2 by antisense survivin RNA in vitro

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective: To study induction of apoptosis and inhibition of proliferation in Hep-2 by antisense survivin RNA. Methods: Antisense survivin RNA expression vector was constructed and then was transfected to human laryngeal carcinoma cell line Hep-2 by lipofectamine. HpEGFP/survivin cells (transfected with the combinant of antisense survivin RNA) were obstained by using G418. The levels of survivin protein before and after transfection were determined by Western-blot. Proliferation activity was measured by MTT assay. The experiment of colony formation in soft agar was carried out for assessing ability of proliferation of Hep-2 cell. Apoptosis was assessed by flow cytometry and acrdine orange(AO).Results:After antisense survivin RNA plasmids were transfected, the level of survivin protein was inhibited in Hep-2. ComPared with control, proliferation of HpEGFP/survivin cells were suppressed significantly. The experiment of colony formation in soft agar showed the ability of colony formation decreased in HpEGFP/survivin cells compared to control (P<0.05). Apoptosis rate increased about 1.81 folds compared with control. Conclusion: The antisense survivin RNA can partly inhibit the level of survivin protein expression in Hep-2 and can induce apoptosis and inhibit the proliferation of Hep-2 by down-regulating the expression of endogenous survivin in vitro.

  4. Latency-Associated Nuclear Antigen of Kaposi's Sarcoma-Associated Herpesvirus (KSHV) Upregulates Survivin Expression in KSHV-Associated B-Lymphoma Cells and Contributes to Their Proliferation▿

    OpenAIRE

    Lu, Jie; Verma, Subhash C.; Murakami, Masanao; Cai, Qiliang; KUMAR, Pankaj; Xiao, Bingyi; Robertson, Erle S.

    2009-01-01

    Survivin is a master regulator of cell proliferation and cell viability and is highly expressed in most human tumors. The molecular network linked to survivin expression in tumors has not been completely elucidated. In this study, we show that latency-associated nuclear antigen (LANA), a multifunctional protein of Kaposi's sarcoma-associated herpesvirus (KSHV) that is found in Kaposi's sarcoma tumors, upregulates survivin expression and increases the proliferation of KSHV-infected B cells. An...

  5. IFN-γ upregulates survivin and Ifi202 expression to induce survival and proliferation of tumor-specific T cells.

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    Mary Zimmerman

    Full Text Available BACKGROUND: A common procedure in human cytotoxic T lymphocyte (CTL adoptive transfer immunotherapy is to expand tumor-specific CTLs ex vivo using CD3 mAb prior to transfer. One of the major obstacles of CTL adoptive immunotherapy is a lack of CTL persistence in the tumor-bearing host after transfer. The aim of this study is to elucidate the molecular mechanisms underlying the effects of stimulation conditions on proliferation and survival of tumor-specific CTLs. METHODOLOGY/PRINCIPAL FINDINGS: Tumor-specific CTLs were stimulated with either CD3 mAb or cognate Ag and analyzed for their proliferation and survival ex vivo and persistence in tumor-bearing mice. Although both Ag and CD3 mAb effectively induced the cytotoxic effecter molecules of the CTLs, we observed that Ag stimulation is essential for sustained CTL proliferation and survival. Further analysis revealed that Ag stimulation leads to greater proliferation rates and less apoptosis than CD3 mAb stimulation. Re-stimulation of the CD3 mAb-stimulated CTLs with Ag resulted in restored CTL proliferative potential, suggesting that CD3 mAb-induced loss of proliferative potential is reversible. Using DNA microarray technology, we identified that survivin and ifi202, two genes with known functions in T cell apoptosis and proliferation, are differentially induced between Ag- and CD3 mAb-stimulated CTLs. Analysis of the IFN-γ signaling pathway activation revealed that Ag stimulation resulted in rapid phosphorylation of STAT1 (pSTAT1, whereas CD3 mAb stimulation failed to activate STAT1. Chromatin immunoprecipitation revealed that pSTAT1 is associated with the promoters of both survivin and ifi202 in T cells and electrophoresis mobility shift assay indicated that pSTAT1 directly binds to the gamma activation sequence element in the survivin and ifi202 promoters. Finally, silencing ifi202 expression significantly decreased T cell proliferation. CONCLUSIONS/SIGNIFICANCE: Our findings delineate a new

  6. Down-regulation of Survivin by Antisense Oligonucleotides Increases Apoptosis, Inhibits Cytokinesis and Anchorage-Independent Growth

    Directory of Open Access Journals (Sweden)

    Jun Chen

    2000-05-01

    Full Text Available Survivin, a member of the inhibitor of apoptosis protein (IAP family, is detected in most common human cancers but not in adjacent normal cells. Previous studies suggest that survivin associates with the mitotic spindle and directly inhibits caspase activity. To further investigate the function of survivin, we used a survivin antisense (AS oligonucleotide to downregulate survivin expression in normal and cancer cells. We found that inhibition of survivin expression increased apoptosis and polyploidy while decreasing colony formation in soft agar. Immunohistochemistry showed that cells without survivin can initiate the cleavage furrow and contractile ring, but cannot complete cytokinesis, thus resulting in multinucleated cells. These findings indicate that survivin plays important roles in a late stage of cytokinesis, as well as in apoptosis.

  7. Cytoplasmic localization of wild-type survivin is associated with constitutive activation of the PI3K/Akt signaling pathway and represents a favorable prognostic factor in patients with acute myeloid leukemia.

    Science.gov (United States)

    Serrano-López, Juana; Serrano, Josefina; Figueroa, Vianihuini; Torres-Gomez, Antonio; Tabares, Salvador; Casaño, Javier; Fernandez-Escalada, Noemi; Sánchez-Garcia, Joaquín

    2013-12-01

    Survivin is over-expressed in most hematologic malignancies but the prognostic significance of the subcompartmental distribution of wild-type or splicing variants in acute myeloid leukemia has not been addressed yet. Using western blotting, we assessed the expression of wild-type survivin and survivin splice variants 2B and Delta-Ex3 in nuclear and cytoplasmic protein extracts in samples taken from 105 patients at the time of their diagnosis of acute myeloid leukemia. Given that survivin is a downstream effector of the PI3K/Akt signaling pathway, survivin expression was also correlated with pSer473-Akt. Wild-type survivin and the 2B splice variant were positive in 76.3% and 78.0% of samples in the nucleus, cytoplasm or both, whereas the Delta-Ex3 isoform was only positive in the nucleus in 37.7% of samples. Cytoplasmic localization of wild-type survivin was significantly associated with the presence of high levels of pSer473-Akt (P<0.001). Inhibition of the PI3K/Akt pathway with wortmannin and Ly294002 caused a significant reduction in the expression of cytoplasmic wild-type survivin. The presence of cytoplasmic wild-type survivin and pSer473-Akt was associated with a lower fraction of quiescent leukemia stem cells (P=0.02). The presence of cytoplasmic wild-type survivin and pSer473-Akt were favorable independent prognostic factors. Moreover, the activation of the PI3K/Akt pathway with expression of cytoplasmic wild-type survivin identified a subgroup of acute myeloid leukemia patients with an excellent outcome (overall survival rate of 60.0±21.9% and relapse-free survival of 63.0±13.5%). Our findings suggest that cytoplasmic wild-type survivin is a critical downstream effector of the PI3K/Akt pathway leading to more chemosensitive cells and a more favorable outcome in acute myeloid leukemia.

  8. Nuclear accumulation of Yes-Associated Protein (YAP) maintains the survival of doxorubicin-induced senescent cells by promoting survivin expression.

    Science.gov (United States)

    Ma, Kai; Xu, Qing; Wang, Shuren; Zhang, Weina; Liu, Mei; Liang, Shufang; Zhu, Hongxia; Xu, Ningzhi

    2016-05-28

    Although chemotherapeutic drugs can induce senescence to prohibit further division of tumor cells, senescence could also promote tumorigenesis mainly through a senescence-associated secretory phenotype. Therefore, senescent tumor cells should be eliminated immediately to prevent drug resistance and recurrence. Here, we used a doxorubicin-induced senescence model to explore the mechanism underlying the survival of therapy-induced senescent cells. After low-dose doxorubicin treatment, tumor cells turned on a senescence program and became large and flattened, increasing their contact area with the extracellular matrix (ECM). Furthermore, Yes-associated protein (YAP) accumulated in the nucleus and YAP activity was increased in doxorubicin-induced senescent cells. Knockdown of YAP increased the sensitivity of cells to low-dose doxorubicin treatment, causing apoptosis rather than senescence. Moreover, the anti-apoptotic gene survivin, a YAP target gene, was overexpressed in senescent cells. Inhibition of survivin could lead to selective elimination of senescent cells through apoptosis. Our study indicates that nuclear accumulation of YAP could promote the survival of senescent cells by increasing survivin expression. Therefore, targeting YAP or survivin might be a new strategy for clearing senescent cancer cells during drug treatment.

  9. Heavy Ion Beams Induce Survivin Expression in Human Hepatoma SMMC-7721 Cells More Effectively than X-rays

    Institute of Scientific and Technical Information of China (English)

    Li GONG; Xiaodong JIN; Qiang LI; Jiangtao LIU; Lizhe AN

    2007-01-01

    High linear energy transfer (LET) heavy ion radiation is more effective in inducing biological damage than low-LET X-rays or γ-rays. Heavy ion beam provides good dose localization (Bragg peak) in critical cancer tissue and gives higher relative biological effectiveness in cell killing across the dose peak, so high-LET heavy ion beam is superior to low-LET radiation in cancer treatment. Survivin, as a member of the inhibitor of apoptosis protein family, might help cancerous cells to overcome the G2/M apoptotic checkpoint and favor the aberrant progression of transformed cells through mitosis. Survivin expression in the human hepatoma SMMC-7721 cell line after exposure to low-LET X-ray and high-LET carbon ion irradiation was investigated in this study. Compared with X-ray irradiation, the carbon ion beam clearly caused G2/M arrest and promoted the expression of the survivin gene in a dose-dependent manner. Clonogenic survival assay showed that SMMC-7721 cells were more radiosensitive to the high-LET carbon ions than to the X-rays, and the radiosensitivity was promoted after treatment with specific survivin short interfering RNA. Differential survivin expression at both transcriptional and translational levels was found for SMMC-7721 cells following low- and high-LET irradiation. The overexpression of survivin in SMMC-7721 cells is probably an important reason why the cancerous cells have radioresistance to strong stimulus such as dense ionizing high-LET radiation. However, the direct killing effect on cancerous cells by high-LET radiation might be more significant than the apoptosis inhibition through the overexpression of survivin following heavy ion irradiation.

  10. EGFR signaling promotes β-cell proliferation and survivin expression during pregnancy.

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    Elina Hakonen

    Full Text Available Placental lactogen (PL induced serotonergic signaling is essential for gestational β-cell mass expansion. We have previously shown that intact Epidermal growth factor -receptor (EGFR function is a crucial component of this pathway. We now explored more specifically the link between EGFR and pregnancy-induced β-cell mass compensation. Islets were isolated from wild-type and β-cell-specific EGFR-dominant negative mice (E1-DN, stimulated with PL and analyzed for β-cell proliferation and expression of genes involved in gestational β-cell growth. β-cell mass dynamics were analyzed both with traditional morphometrical methods and three-dimensional optical projection tomography (OPT of whole-mount insulin-stained pancreata. Insulin-positive volume analyzed with OPT increased 1.4-fold at gestational day 18.5 (GD18.5 when compared to non-pregnant mice. Number of islets peaked by GD13.5 (680 vs 1134 islets per pancreas, non-pregnant vs. GD13.5. PL stimulated beta cell proliferation in the wild-type islets, whereas the proliferative response was absent in the E1-DN mouse islets. Serotonin synthesizing enzymes were upregulated similarly in both the wild-type and E1-DN mice. However, while survivin (Birc5 mRNA was upregulated 5.5-fold during pregnancy in the wild-type islets, no change was seen in the E1-DN pregnant islets. PL induced survivin expression also in isolated islets and this was blocked by EGFR inhibitor gefitinib, mTOR inhibitor rapamycin and MEK inhibitor PD0325901. Our 3D-volumetric analysis of β-cell mass expansion during murine pregnancy revealed that islet number increases during pregnancy. In addition, our results suggest that EGFR signaling is required for lactogen-induced survivin expression via MAPK and mTOR pathways.

  11. EFFECT OF STRESS ON THE PERCENT BODY WEIGHT CHANGE AND MRNA EXPRESSION OF IGF-1, SURVIVINE AND HSP-70 GENE IN THE HIERARCHIAL FOLLICLES OF JAPANESE QUAIL

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    N Shit

    2014-12-01

    Full Text Available The present study was carried out to explore the effect of stress on body weight and the mRNA expression of IGF-1, Survivine and HSP-70 gene in the hierarchial follicles of Japanese quail. A total 24 birds (10 weeks were taken and stress was induced by immobilization daily for 2hrs (between 9.00 - 11.00 AM throughout the study (10 days. Four birds were sacrificed on 1, 2, 4, 6, 8 and 10 days of the treatment. Hierarchial follicles (F1, F2 & F3 were aseptically collected to quantify the expression of IGF-1, Survivine and HSP-70 gene using real-time PCR technique. The percent body weight reduction increased and reached highest (21.30% on 10th day. The fold expression of IGF-1 gene was significantly ((P=0.05 down regulated in advance to the time of experiment. However, the fold expression of survivine gene was significantly (P=0.05 up regulated and the intensity was highest (17 fold in F-3 follicle on 4th day of experiment. No significant change in the mRNA expression of HSP-70 gene was evident in this study. From this study it may be concluded that stress brings physio-molecular change through HPA activation, which not only causes tissue regression also modifies the cellular mechanism.

  12. STAT1 and Survivin Expression in Full Lymph Node Examined Gastric Cancer by Using Tissue Microarray Technique

    Institute of Scientific and Technical Information of China (English)

    DENG Hao; WU Renliang; CHEN Ying; LIU Lijiang

    2006-01-01

    Objective: To characterize the relationship between STAT1 and Survivin expression, and the relationship between them and lymph node metastasis, depth of invasion and prognosis in full lymph node examined gastric cancer patients of China. Methods: Specimens of curative dissection between 1988 and 2003 were collected from the affiliated hospital of Jianghan University. All 140 patients had complete examination data. All lymph nodes were found by clearing fat method. The interrupted serial 4 μm sections, routine hematoxylin and eosin staining and immunohistochemical methods were used to detect the lymph node metastases. Gastric cancer tissue microarray was formed and the expression of survivin and STAT1 in gastric cancer was detected by immunohistochemical method. All data were processed using Spearman rank correlation analysis, Kaplan-Meyer Log-rank method and Cox multivariate analysis (SPSS12.0 software). Results: Among 140 gastric cancer tissue microarrays constructed, 110 could be used(utilization rate was 78.6%). 7079 lymph nodes were found in 110 cases (64.4/case). Metastases were found in 89 cases and 1679 lymph nodes. Positive expression rate of survivin and STAT1 was 52.7% (58/110)and 40% (44/110) respectively. There was a significant negative correlation between STAT1 expression and survivin expression (r=-0.19, P=-0.04). STAT1 expression had a negative correlation with depth of invasion(r=-0.21, P=0.04). Survivin expression had a negative correlation with UICC N stage (r=-0.24, P=0.01)and histological classification (r=-0.21, P=0.03) by Spearman rank correlation analysis. But survivin and STAT1 expression was not related with prognosis. A significant correlation between lymph node metastasis and prognosis was demonstrated by Cox multivariate analysis (χ2=4.85, P=0.028). Conclusion: STAT1 has a negative correlation with survivin expression in gastric cancer. Both of them have no correlation with prognosis in gastric cancer. STAT1 expression can be a

  13. The study of neurogenesis contributed by the expression of survivin in hippocampus after traumatic brain injury%脑创伤后海马区生存素蛋白促进神经再生的研究

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    张琳; 颜荣; 刘晓智; 赵玉军; 陈镭

    2015-01-01

    (42 003.15,P <0.05) and lasting to day 7,peaking at day 2 (90 403.34,P < 0.01).The number of survivin +,BrdU + and doublecortin (DCX +) cells in subgranular zone (SGZ) of ipsilateral hippocampus was significantly increased after TBI by immunofluorescence staining.The results also revealed that the majority of survivin + cells were BrdU + cells,and a part of survivin + cells were DCX + cells in SGZ of dentate gyrus (DG) in the hippocampus.Conclusion Survivin gene was activated follow brain injury.The expression of survivin protein were increased in the hippocampus after TBI.Increased survivin protein is expressed in neural stem cells and immature neurons in ipsilateral hippocampus after TBI,which correlate to the proliferation of neurogenic cell in SGZ of hippocampus.

  14. Study of the Expression of Survivin & Its Splice Variants; ΔEx3, 2b and 3b as Diagnostic Molecular Markers in Breast Cancer

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    E Babaei

    2009-07-01

    Full Text Available Introduction: Survivin is a new member of the Inhibitor Apotosis Protein family (IAP which plays an important role in the regulation of both cell cycle and apoptosis. Its distinct expression in tumor cells as compared to normal adult cells introduces Survivin as the fourth transcriptom demonstrated in tumors. Breast cancer is the most common malignancy among women and scientist`s efforts to classify it has lead to various molecular subtypes and controversial results. Because of the high prevalence of these tumors and lack of suitable molecular markers for diagnosis and prognosis, there are ongoing efforts to find molecular markers which can distinguish nontumoral from tumor tissues. In this study we evaluate the potential usefulness of Survivin and its splice variants ΔEx3, 2b and 3b as molecular markers in breast cancer. Methods: We studied 18 tumor and 17 non tumor adjacent tissues. Transcription levels were measured by Semiquantitative Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR and normalized by ß2m as an internal control. Results: 1Survivin and its splice variants; Δex3, 2b and 3b showed differentially higher expression levels in tumors than adjacent normal tissues. 2 The expression levels of Survivin, Survivin-ΔEx3 and Survivin-3b were significantly correlated with the type of tumors. 3 Survivin-2b was expressed in a few samples. 4 Survivin-3b was detected only in tumor samples. Also, our results showed that ΔEx3 variant can be introduced as a dominant expressed variant in breast cancer. Conclusion: Our data indicated that the expression of Survivin, Survivin ∆Ex3 and especially, Survivin-3b were correlated with cancerous nature of tumors and Survivin-∆Ex3 was the most common expressed variant in breast carcinomas. These results besides confirming the potential usefulness of Survivin and its splice variants as molecular markers in breast cancer, demonstrated the role of the gene and its splice variants, especially 3b

  15. Changes of activated circulating endothelial cells and survivin in patients with non-small cell lung cancer after antiangiogenesis therapy

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    WANG Jing; HUANG Chun; WEI Xi-yin; QI Da-liang; GONG Li-qun; MU Hai-yu; YAO Qiang; LI Kai

    2008-01-01

    Background Although antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect,there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy.This study was undertaken to investigate the changes of activated circulating endothelial cells (aCECs) and survMn after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy.Methods Patients of non-small cell lung cancer (NSCLC) treated with chemotherapy with or without Endostar were observed.The amount of activated CECs was detected by flow cytometry,and the expression of survivin mRNA was determined by real-time polymerase chain reaction (PCR).Results After treatment,the amount of activated CECs decreased significantly in clinical benefit cases (P=-0.021 in chemotherapy alone,P=0.001 in chemotherapy plus Endostar),increased in disease progressive cases (P=-0.015 in chemotherapy alone,but P=0.293 in chemotherapy with Endotatar).After therapy,the expression of survivin mRNA decreased in clinical benefit cases (P=0.001) and increased in disease progressive cases (P=0.018).A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy (P=0.001 and 0.021,respectively),but only in the chemotherapy with Endostar group pre-therapy (P=0.030) rather than post-therapy.A positive correlation was found between the decreased activated CECs after therapy and time to progression (TTP) (r=0.322,P=0.012);a negative correlation was found between the amount of survivin mRNA in serum post-therapy and TTP(r= -0.291,P=0.048).Conclusions Activated CECs and survMn may be ideal markers forecasting efficacy and prognosis of NSCLC.The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors.Their combination can evaluate more accurately the efficacy of antiangiogenic

  16. EM011 activates a survivin-dependent apoptotic program in human non-small cell lung cancer cells

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    Yates Clayton

    2009-10-01

    Full Text Available Abstract Background Lung cancer remains a leading cause of cancer death among both men and women in the United States. Treatment modalities available for this malignancy are inadequate and thus new drugs with improved pharmacological profiles and superior therapeutic indices are being continually explored. Noscapinoids constitute an emerging class of anticancer agents that bind tubulin but do not significantly alter the monomer/polymer ratio of tubulin. EM011, a rationally-designed member of this class of non-toxic agents, is more potent than the lead molecule, noscapine. Results Here we report that EM011 inhibited proliferation of a comprehensive panel of lung cancer cells with IC50's ranging from 4-50 μM. In A549 human non-small cell lung cancer cells, the antiproliferative activity was mediated through blockage of cell-cycle progression by induction of a transient but robust mitotic arrest accompanied by activation of the spindle assembly checkpoint. The mitotically-arrested A549 cells then override the activated mitotic checkpoint and aberrantly exit mitosis without cytokinesis resulting in pseudo G1-like multinucleated cells that either succumb directly to apoptosis or continue another round of the cell-cycle. The accumulated enormous DNA perhaps acts as genotoxic stress to trigger cell death. EM011-induced apoptotic cell death in A549 cells was associated with a decrease of the Bcl2/BAX ratio, activation of caspase-3 and cleavage of PARP. Furthermore, EM011 induced downregulation of survivin expression over time of treatment. Abrogation of survivin led to an increase of cell death whereas, overexpression caused decreased apoptosis. Conclusion These in vitro data suggest that EM011 mediates antiproliferative and proapoptotic activity in non-small cell A549 lung cancer cells by impeding cell-cycle progression and attenuating antiapoptotic signaling circuitries (viz. Bcl2, survivin. The study provides evidence for the potential usefulness of

  17. Deptor enhances triple-negative breast cancer metastasis and chemoresistance through coupling to survivin expression.

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    Parvani, Jenny G; Davuluri, Gangarao; Wendt, Michael K; Espinosa, Christine; Tian, Maozhen; Danielpour, David; Sossey-Alaoui, Khalid; Schiemann, William P

    2015-03-01

    Transforming growth factor-β (TGF-β) functions to suppress tumorigenesis in normal mammary tissues and early-stage breast cancers and, paradoxically, acts to promote the metastasis and chemoresistance in late-stage breast cancers, particularly triple-negative breast cancers (TNBCs). Precisely how TGF-β acquires oncogenic characteristics in late-stage breast cancers remains unknown, as does the role of the endogenous mammalian target of rapamycin (mTOR) inhibitor, Dep domain-containing mTOR-interacting protein (Deptor), in coupling TGF-β to TNBC development and metastatic progression. Here we demonstrate that Deptor expression was downregulated in basal-like/TNBCs relative to their luminal counterparts. Additionally, Deptor expression was 1) inversely correlated with the metastatic ability of human (MCF10A) and mouse (4T1) TNBC progression series and 2) robustly repressed by several inducers of epithelial-mesenchymal transition programs. Functional disruption of Deptor expression in 4T07 cells significantly inhibited their proliferation and organoid growth in vitro, as well as prevented their colonization and tumor formation in the lungs of mice. In stark contrast, elevated Deptor expression was significantly associated with poorer overall survival of patients harboring estrogen receptor α-negative breast cancers. Accordingly, enforced Deptor expression in MDA-MB-231 cells dramatically enhanced their 1) organoid growth in vitro, 2) pulmonary outgrowth in mice, and 3) resistance to chemotherapies, an event dependent on the coupling of Deptor to survivin expression. Collectively, our findings highlight the dichotomous functions of Deptor in modulating the proliferation and survival of TNBCs during metastasis; they also implicate Deptor and its stimulation of survivin as essential components of TNBC resistance to chemotherapies and apoptotic stimuli.

  18. Deptor Enhances Triple-Negative Breast Cancer Metastasis and Chemoresistance through Coupling to Survivin Expression

    Directory of Open Access Journals (Sweden)

    Jenny G. Parvani

    2015-03-01

    Full Text Available Transforming growth factor–β (TGF-β functions to suppress tumorigenesis in normal mammary tissues and early-stage breast cancers and, paradoxically, acts to promote the metastasis and chemoresistance in late-stage breast cancers, particularly triple-negative breast cancers (TNBCs. Precisely how TGF-β acquires oncogenic characteristics in late-stage breast cancers remains unknown, as does the role of the endogenous mammalian target of rapamycin (mTOR inhibitor, Dep domain–containing mTOR-interacting protein (Deptor, in coupling TGF-β to TNBC development and metastatic progression. Here we demonstrate that Deptor expression was downregulated in basal-like/TNBCs relative to their luminal counterparts. Additionally, Deptor expression was 1 inversely correlated with the metastatic ability of human (MCF10A and mouse (4T1 TNBC progression series and 2 robustly repressed by several inducers of epithelial-mesenchymal transition programs. Functional disruption of Deptor expression in 4T07 cells significantly inhibited their proliferation and organoid growth in vitro, as well as prevented their colonization and tumor formation in the lungs of mice. In stark contrast, elevated Deptor expression was significantly associated with poorer overall survival of patients harboring estrogen receptor α–negative breast cancers. Accordingly, enforced Deptor expression in MDA-MB-231 cells dramatically enhanced their 1 organoid growth in vitro, 2 pulmonary outgrowth in mice, and 3 resistance to chemotherapies, an event dependent on the coupling of Deptor to survivin expression. Collectively, our findings highlight the dichotomous functions of Deptor in modulating the proliferation and survival of TNBCs during metastasis; they also implicate Deptor and its stimulation of survivin as essential components of TNBC resistance to chemotherapies and apoptotic stimuli.

  19. Dynamic intracellular survivin in oral squamous cell carcinoma: underlying molecular mechanism and potential as an early prognostic marker.

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    Engels, K; Knauer, S K; Metzler, D; Simf, C; Struschka, O; Bier, C; Mann, W; Kovács, A F; Stauber, R H

    2007-04-01

    Survivin functions as an apoptosis inhibitor and a regulator of cell division in many tumours. The intracellular localization of survivin in tumours has been suggested as a prognostic marker. However, current reports are inconsistent and the underlying molecular mechanisms are not understood. The present study has examined the localization and prognostic value of nuclear and cytoplasmic survivin in the pre-therapeutic biopsies from 71 oral and oropharyngeal squamous carcinoma (OSCC) patients. Statistical analysis indicated that preferential nuclear versus cytoplasmic survivin correlated with favourable versus unfavourable disease outcome. Uni- and multi-variate analysis showed that in contrast to total survivin expression, the difference between nuclear and cytoplasmic survivin was a strong predictor for relapse-free survival (p=0.0003). As a potential underlying molecular mechanism, it is shown in OSCC cell lines that predominantly cytoplasmic survivin mediates protection against chemo- and radio-therapy-induced apoptosis. Importantly, the cytoplasmic localization of survivin is regulated by its nuclear export signal (NES), and export-deficient nuclear survivin is not cytoprotective. This study suggests that the difference between cytoplasmic and nuclear survivin is an indicator for survivin activity in tumour cells. Thus, this difference may serve as a predictive marker of outcome in OSCC patients undergoing multi-modality therapy. The pharmacogenetic interference with survivin's cytoplasmic localization is also to be pursued as a potential therapeutic strategy.

  20. High expression of nuclear survivin and Aurora B predicts poor overall survival in patients with head and neck squamous cell cancer

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    Erpolat, O.P.; Akmansu, M. [Medical School of Gazi Univ., Besevler-Ankara (Turkey). Dept. of Radiation Oncology; Gocun, P.U.; Karakus, E.; Akyol, G. [Medical School of Gazi Univ., Besevler-Ankara (Turkey). Dept. of Pathology

    2012-03-15

    Survivin is one of the apoptosis inhibitor proteins. Together with Aurora B, it also plays a role in regulating several aspects of mitosis. High expression of these markers is correlated with malignant behavior of various cancers and resistance to therapy. Our aim was to evaluate the prognostic role of these markers in head and neck cancers. We evaluated the expression of Aurora B and survivin in tissue specimens of 58 patients with head and neck squamous cell carcinoma using immunohistochemistry. Patients who showed high expression of cytoplasmic and nuclear survivin and Aurora B had significantly shorter overall survival (p = 0.036, p < 0.000, p = 0.032, respectively). In multivariate analysis, high expression of nuclear survivin was the only independent negative prognostic factor (p = 0.024). Moreover, it was found that high co-expression of nuclear survivin and Aurora B had a negative effect on survival in univariate (p < 0.000) and multivariate (p < 0.000) analyses. The negative prognostic values of high expression of Aurora B and high co-expression of nuclear survivin and Aurora B on survival were shown. These findings suggest that co-expression of nuclear survivin and Aurora B can be useful diagnostic markers and therapeutic targets for head and neck squamous cell carcinoma. However, further studies with a larger number of patients in a more homogeneous disease group are needed to confirm the conclusion.

  1. Effect of all-trans retinoic acid 0n drug sensitivity and expression of survivin in LoVo cells

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Background All-trans retinoic acid(ATRA)can influence the tumor cell proliferation cycle,and some chemotherapeutic drugs are cycle specific.In this study,we hypothesize that ATRA can enhance chemotherapeutic drug sensitivity by affecting the cell cycle of tumor cells.Methods The cell cycle of LoVo cells was evaluated using flow cytometry(FCM).Cell viability was analyzed using the MTT assay.The morphologic changes in the treated LoVo cells were measured with acridine orange (AO)/ethidium bromide(EB)staining.Expression of survivin in LoVo cells was analyzed by immunofluorescence assay.Results After LoVo cells were treated with ATRA,the G0/G1 ratio of the tumor cells increased and the cell ratio of Sand G2/M-phase decreased.Viability of the cells decreased significantly after combined treatment with ATRA and 5-fluorouracil(5-FU)or mitomycin c(MMC) and was evaluated by fluorescence microscopy.Expression level of survivin in the tumor cells decreased after ATRA combination treatment.Conclusions ATRA enhances drug sensitivity of the LoVo cell line to cell cycle-specific agents and inhibits the expression of survivin in LoVo cells.The combination of ATRA and 5-FU or MMC promoted cell apoptosis,and the mechanism involved in apoptosis may be related to inhibition of survivin gene expression.

  2. Expression of Survivin, CyclinD1, p21WAF1, Caspase-3 in Cervical Cancer and Its Relation with Prognosis

    Institute of Scientific and Technical Information of China (English)

    LU Shi; ZHANG Baohua; WANG Zehua

    2005-01-01

    The implications of Survivin, CyclinD1, p21WAF1, Caspase-3 in the development, progression and prognosis in cervical cancer were investigated. By using immunohistochemical SP method, the expression of Survivin, CyclinD1, p21WAF1 , Caspase-3 was detected in 41 cases of cervical cancer, 17 cases of cervical intraepithelial neoplasia (CIN) and 10 cases of normal tissues, and their relation with pathological grade, clinical stage, metastasis and survival time was analyzed.The results showed that the positive expression rate of Survivin, CyclinD1 in cervical cancer was significantly higher than in CIN group and normal control group (P<0.05). The median survival time in the patients with cervical cancer positive for Survivin and CyclinD1 was significantly shorter than in those with negative expression (P<0.05). The expression of both Survivin and CyclinD1 was not related with tumor grade, clinical stage and metastasis (P>0. 05). The positive expression rate of p21WAF1 , Caspase-3 in cervical ca rcer was significantly lower than in CIN group and normal control group (P<0.05), and had a close relation with tumor grade (P<0.05). The expression of Survivin in cervical cancer in cervical cancer was negatively associated with that of Caspase-3 (P<0.01), but positively with that of CyclinD1 (P<0.01). Cox Multivariate analysis revealed that Survivin was the independent prognostic indicator influencing the survival time of the patients with cervical cancer (P<0.05). It was suggested that the high expression of Survivin or CyclinD1, and low expression of p21WAF1 or Caspase-3 was closely correlated with the development of cervical cancer. Survivin and CyclinD1 could be used as a useful indicator to predict the prognosis of cervical cancer.

  3. Detection of visfatin, Xiap and Survivin expressions in placenta tissue of preeclampsia and its correlation with serum indexes

    Institute of Scientific and Technical Information of China (English)

    Wei Zhong

    2015-01-01

    Objective: To study the expressions of visfatin, Xiap and Survivin in placenta tissue of preeclampsia and its correlation with serum indexes. Methods: Preeclampsia patients who gave birth in our hospital and healthy volunteers during the same period were selected and enrolled in observation group and control group. Then placenta tissue was collected and mRNA contents of visfatin, Xiap and Survivin were detected; serum was collected and angiogenesis related factors, inflammatory cytokines were detected. Results: (1) Placenta index: compared with mRNA contents of target genes in placenta tissue of control group, mRNA content of Visfatin in placenta tissue of observation group was higher; mRNA contents of Xiap and Survivin were lower; (2) Serum angiogenesis related factors: compared with contents of serum angiogenesis cytokines of control group, serum PIGF and Glycodelin contents of observation group were lower; sEng, sFlt-1, PP13 and HtrA1 contents were higher; (3) Inflammatory cytokines: compared with serum inflammation related factor contents of control group, serum YKL-40, CXCL-10, Chemerin, IL-18, HMGB-1 and MIF contents of observation group were higher. Conclusion: Abnormal expressions of visfatin, Xiap and Survivin in placenta tissue are related to the occurrence of preeclampsia, and gene mRNA contents are related to the contents of serum angiogenesis related factors and inflammatory cytokines.

  4. Development of survivin and tumor research

    Institute of Scientific and Technical Information of China (English)

    JI Yu-bin; LIU Guang-da; YU Lei; LI Hai-jiao; YANG Hai-fan; PANG Lin-lin

    2008-01-01

    Survivin was firstly separated in hybridization of Effector Cell Protease Receptor-1(EPR-1) cDNA in human genome by Yale University's Ambrosini in 1997, which is member of the inhibitor of apoptosis proteins (IAPS). Unlike other IAP protein, found during embryonic and fetal development, survivin wascompletely down-regulated and undetectable in normal adult tissues, and became prominentlyre expressed in all of the most common cancers. It through includes the cysteine/histidine rod-shaped viral IAP repetition sequence baculoviral IAP repeats(BIRs) the structure territory directly or intervenes Caspases the function to display indirectly its anti-perishes weakly the function, simultaneously it also is in the cell division process the chromosome traveler protein (chromosome passenger protein). There are three approaches by which survivin inhibits the processing of apoptosis: (1)inhibits processing of down stream effector caspase-3, caspase-7and caspase-9 in cell receiving apoptotic stimuls; (2)with the Smac/DIABLO function, sends the XIAP activeness to increase, XIAP through directly affects and restrains its function with caspases, achieved restrains function which perishes weakly; (3) through restrains p53 the function to block perishes weakly the process. Survivin expressed specificity and its function multiplicity. Survivin only expresses in tumor tissues and cannot be found in normal terminally differentiated tissues. This kind of expression is been extremely low the cell cycle strict regulation in the G1 time expression, the S time is G1 time 6 times, the G2/M time advances to 40 times, demonstrated its expression has the G2/M time dependence specificity. It has bi-function of inhibiting apoptosis and involving in cell cycle control. Survivin has found in most of tumor ceils in recently researches. Survivin expresses generally in all tumor cell line in the American State-run Cancer Research institute anti-tumor medicine screening procedure 60 different tumor

  5. Brain targeted PLGA nanocarriers alleviating amyloid-Β expression and preserving basal survivin in degenerating mice model.

    Science.gov (United States)

    Sriramoju, Bhasker; Neerati, Prasad; Kanwar, Rupinder K; Kanwar, Jagat R

    2015-11-01

    The chronic systemic administration of d-Galactose in C57BL/6J mice showed a relatively high oxidative stress, amyloid-β expression and neuronal cell death. Enhanced expression of pyknotic nuclei, caspase-3 and reduced expression of neuronal integrity markers further confirmed the aforesaid insults. However, concomitant treatment with the recombinant protein (SurR9-C84A) and the anti-transferrin receptor antibody conjugated SurR9-C84A (SurR9+TFN) nanocarriers showed a significant improvement in the disease status and neuronal health. The beauty of this study is that the biodegradable Food and Drug Administration (FDA) approved poly(lactic-co-glycolic acid) (PLGA) nanocarriers enhanced the biological half-life and the efficacy of the treatments. The nanocarriers were effective in lowering the amyloid-β expression, enhancing the neuronal integrity markers and maintaining the basal levels of endogenous survivin that is essential for evading the caspase activation and apoptosis. The current study herein reports for the first time that the brain targeted SurR9-C84A nanocarriers alleviated the d-Galactose induced neuronal insults and has potential for future brain targeted nanomedicine application.

  6. Downregulation of survivin expression and concomitant induction of apoptosis by celecoxib and its non-cyclooxygenase-2-inhibitory analog, dimethyl-celecoxib (DMC, in tumor cells in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    Hofman Florence M

    2006-05-01

    Full Text Available Abstract Background 2,5-Dimethyl-celecoxib (DMC is a close structural analog of the selective cyclooxygenase-2 (COX-2 inhibitor celecoxib (Celebrex® that lacks COX-2-inhibitory function. However, despite its inability to block COX-2 activity, DMC is able to potently mimic the anti-tumor effects of celecoxib in vitro and in vivo, indicating that both of these drugs are able to involve targets other than COX-2 to exert their recognized cytotoxic effects. However, the molecular components that are involved in mediating these drugs' apoptosis-stimulatory consequences are incompletely understood. Results We present evidence that celecoxib and DMC are able to down-regulate the expression of survivin, an anti-apoptotic protein that is highly expressed in tumor cells and known to confer resistance of such cells to anti-cancer treatments. Suppression of survivin is specific to these two drugs, as other coxibs (valdecoxib, rofecoxib or traditional NSAIDs (flurbiprofen, indomethacin, sulindac do not affect survivin expression at similar concentrations. The extent of survivin down-regulation by celecoxib and DMC in different tumor cell lines is somewhat variable, but closely correlates with the degree of drug-induced growth inhibition and apoptosis. When combined with irinotecan, a widely used anticancer drug, celecoxib and DMC greatly enhance the cytotoxic effects of this drug, in keeping with a model that suppression of survivin may be beneficial to sensitize cancer cells to chemotherapy. Remarkably, these effects are not restricted to in vitro conditions, but also take place in tumors from drug-treated animals, where both drugs similarly repress survivin, induce apoptosis, and inhibit tumor growth in vivo. Conclusion In consideration of survivin's recognized role as a custodian of tumor cell survival, our results suggest that celecoxib and DMC might exert their cytotoxic anti-tumor effects at least in part via the down-regulation of survivin – in a

  7. SDF-1/CXCR4 Axis Regulates Cell Cycle Progression and Epithelial-Mesenchymal Transition via Up-regulation of Survivin in Glioblastoma.

    Science.gov (United States)

    Liao, Anyan; Shi, Ranran; Jiang, Yuliang; Tian, Suqing; Li, Panpan; Song, Fuxi; Qu, Yalan; Li, Jinna; Yun, Haiqin; Yang, Xiangshan

    2016-01-01

    Stromal cell-derived factor 1 (SDF-1)/CXCR4 ligand-receptor axis is widely recommended as an attractive target for cancer therapy. Meanwhile, epithelial-mesenchymal transition (EMT) process is linked to disease pathophysiology. As one of inhibitors of apoptosis proteins, survivin is implicated in the onset and development of cancer. In the present study, we tried to determine the cause-effect associations between SDF-1/CXCR4 axis and survivin expression in glioblastoma U-251 cell line. Survivin activation and inhibition were induced with exogenous SDF-1 and survivin small interfering RNA (survivin siRNA), respectively. Western blot was used to detect relevant proteins in SDF-1/CXCR4 axis. Western blot analysis revealed that survivin expression in U-251 increased in a dose- and time-dependent manner in response to SDF-1 treatment. However, the interference with MEK/ERK and PI3K/AKT pathway prohibited SDF-1-induced survivin up-regulation. Importantly, survivin knockdown abrogated cell cycle progression and the expression of snail and N-cadherin, compared with non-transfectants. In conclusion, the present study shows that SDF-1 up-regulates survivin via MEK/ERK and PI3K/AKT pathway, leading to cell cycle progression and EMT occurrence dependent on survivin. The blockade of survivin will allow for the treatment of glioblastoma.

  8. Efficacious gene silencing in serum and significant apoptotic activity induction by survivin downregulation mediated by new cationic gemini tocopheryl lipids.

    Science.gov (United States)

    Kumar, Krishan; Maiti, Bappa; Kondaiah, Paturu; Bhattacharya, Santanu

    2015-02-02

    Nonviral gene delivery offers cationic liposomes as promising instruments for the delivery of double-stranded RNA (ds RNA) molecules for successful sequence-specific gene silencing (RNA interference). The efficient delivery of siRNA (small interfering RNA) to cells while avoiding unexpected side effects is an important prerequisite for the exploitation of the power of this excellent tool. We present here six new tocopherol based cationic gemini lipids, which induce substantial gene knockdown without any obvious cytotoxicity. All the efficient coliposomal formulations derived from each of these geminis and a helper lipid, dioleoylphosphatidylethanolamine (DOPE), were well characterized using physical methods such as atomic force microscopy (AFM) and dynamic light scattering (DLS). Zeta potential measurements were conducted to estimate the surface charge of these formulations. Flow cytometric analysis showed that the optimized coliposomal formulations could transfect anti-GFP siRNA efficiently in three different GFP expressing cell lines, viz., HEK 293T, HeLa, and Caco-2, significantly better than a potent commercial standard Lipofectamine 2000 (L2K) both in the absence and in the presence of serum (FBS). Notably, the knockdown activity of coliposomes of gemini lipids was not affected even in the presence of serum (10% and 50% FBS) while it dropped down for L2K significantly. Observations under a fluorescence microscope, RT-PCR, and Western blot analysis substantiated the flow cytometry results. The efficient cellular entry of labeled siRNA in GFP expressing cells as evidenced from confocal microscopy put forward these gemini lipids among the potent lipidic carriers for siRNA. The efficient transfection capabilities were also profiled in a more relevant fashion while performing siRNA transfections against survivin (an anti-apoptotic protein) which induced substantial apoptosis. Furthermore, the survivin downregulation improved the therapeutic efficacy levels of an

  9. Arctigenin enhances chemosensitivity to cisplatin in human nonsmall lung cancer H460 cells through downregulation of survivin expression.

    Science.gov (United States)

    Wang, Huan-qin; Jin, Jian-jun; Wang, Jing

    2014-01-01

    Arctigenin, a dibenzylbutyrolactone lignan, enhances cisplatin-mediated cell apoptosis in cancer cells. Here, we sought to investigate the effects of arctigenin on cisplatin-treated non-small-cell lung cancer (NSCLC) H460 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and annexin-V/propidium iodide staining were performed to analyze the proliferation and apoptosis of H460 cells. Arctigenin dose-dependently suppressed cell proliferation and potentiated cell apoptosis, coupled with increased cleavage of caspase-3 and poly(ADP-ribose) polymerase. Moreover, arctigenin sensitized H460 cells to cisplatin-induced proliferation inhibition and apoptosis. Arctigenin alone or in combination with cisplatin had a significantly lower amount of survivin. Ectopic expression of survivin decreased cell apoptosis induced by arctigenin (P arctigenin (P arctigenin has a therapeutic potential in combina-tion with chemotherapeutic agents for NSLC.

  10. Ran GTPase protein promotes human pancreatic cancer proliferation by deregulating the expression of Survivin and cell cycle proteins

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    Deng, Lin [State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi 710032 (China); Department of Oncology, Tangdu Hospital, Fourth Military Medical University, Xi’an, Shaanxi 710038 (China); Lu, Yuanyuan; Zhao, Xiaodi; Sun, Yi; Shi, Yongquan; Fan, Hongwei; Liu, Changhao; Zhou, Jinfeng; Nie, Yongzhan; Wu, Kaichun [State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi 710032 (China); Fan, Daiming, E-mail: daimingfan@fmmu.edu.cn [State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi 710032 (China); Guo, Xuegang, E-mail: xuegangguo@126.com [State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi 710032 (China)

    2013-10-18

    Highlights: •Overexpression of Ran in pancreatic cancer was correlated with histological grade. •Downregulation of Ran could induce cell apoptosis and inhibit cell proliferation. •The effects were mediated by cell cycle proteins, Survivin and cleaved Caspase-3. -- Abstract: Ran, a member of the Ras GTPase family, has important roles in nucleocytoplasmic transport. Herein, we detected Ran expression in pancreatic cancer and explored its potential role on tumour progression. Overexpressed Ran in pancreatic cancer tissues was found highly correlated with the histological grade. Downregulation of Ran led to significant suppression of cell proliferation, cell cycle arrest at the G1/S phase and induction of apoptosis. In vivo studies also validated that result. Further studies revealed that those effects were at least partly mediated by the downregulation of Cyclin A, Cyclin D1, Cyclin E, CDK2, CDK4, phospho-Rb and Survivin proteins and up regulation of cleaved Caspase-3.

  11. Low-dose radiation reverses cisplatin resistance in ovarian cancer cells by changing Survivin and Caspase-3 expression

    Institute of Scientific and Technical Information of China (English)

    Qing Dong; Tao Jiang; Donghai Liang; Xiaoran Liu; Hongsheng Yu

    2016-01-01

    Objective Cisplatin (DDP) is the main chemotherapy drug for ovarian cancer. However, ovarian cancer cel s tend to develop cisplatin resistance in the clinical setting. Tumor cel s are sensitive to low-dose radia-tion (LDR). LDR therapy can improve the ef ects of chemotherapy drugs on ovarian cancer, but the underly-ing mechanisms are not clear. In this study, we explored the impact of low-dose radiation on Survivin and Caspase-3 levels in SKOV3/DDP ovarian cancer cel s that are resistant to cisplatin. Methods Cel viability was examined by cel counting kit-8 (CCK-8) assay, and quantitative PCR was used to detect Caspase-3 and Survivin transcript levels. Flow cytometry was used to detect and quantify apoptotic cel s. Results Cel viability was lower when cel s were treated with LDR and cisplatin than when cel s were treated with conventional radiation and cisplatin, or cisplatin alone (P Conclusion LDR reverses cisplatin resistance in SKOV3/DDP cel s, and may do so by suppressing Survivin expression and increasing Caspase-3 expression.

  12. 人肺腺癌细胞株A549中HIF-1α对Survivin的表达调控%Regulation of survivin expression by hypoxia-inducible factor-1α in non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    李伟; 陈余清; 孙艳; 赵成岭; 王效静

    2011-01-01

    Background and purpose: Survivin gene is a unique member of the inhibitor of apoptosis protein (LAP) family. It plays an important role, not only in regulating mitosis but also in inhibiting apoptosis. It is highly expressed in almost all types of human tumors and fetal tissues but rarely detectable in normal adult tissues. High levels of survivin expression have been associated with tumor progression, resistance to radiation and drug treatments and poor survival rates in cancer patients. The current literature contains few reports on the transcriptional regulation of survivin expression in lung cancer. Previous studies have found that there are also 2 putative binding sites for hypoxia-inducible factor- la(HIF- la) in the core promoter region of survivin gene. Survivin promoter-luciferase reporter vectors Pgl3-SVP230-luc have been constructed early. The purpose of this study was to investigate the mechanism of (HIF-la)on transcriptional regulation of survivin in A549 cells by hypoxia. Methods: (l)Double labeling immunofluorescence method was used to detect co-expression of survivin/HIF-lα protein; (2)RT-polymerase chain reaction (RT-PCR) and Western blot was used to examine the level of survivin Mrna and protein in A549 cells transfected by HIF-lα expression plasmid and HIF-lα siRNA; (3)Luciferase activity was detected in A549 cells following cotransfection with Pgl3-SVP230-luc as well as HIF-la expression plasmid or HIF-lα siRNA to value the transcriptional activity of survivin. (4)Electrophoretic mobility shift assay (EMS A) was performed to test the nuclear extract of the A549 cells binding to the r-32P labeled probes containing survivin promoter squences. Results: (l)Survivin/HIF-lα proteins co-expressed in A549 cell; (2)Compared with control groups, the level of survivin Mrna and protein is markedly increased in A549 cells transfected with HIF-lα expression plasmid, but decreased in the HIF-lα siRNA group(P<0.01); (3)The relative activity of Pgl3-SVP

  13. REAL-TIME DETECTION OF SURVIVIN mRNA EXPRESSION IN CERVICAL CANCER CELL LINES USING MOLECULAR BEACON IMAGING

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The initiated growth of human cancer cells of-ten mostly come fromthe abnor mal expression ofgenes.Survivinis anapotosis inhibitor of IAPfami-ly,cloned by Ambrosini in1997usingthe cDNAofeffector cell protease receptor-1(EPR-1),and is thekey gene for the development and advancement oftumor.Inthe present study,the feasibility of detec-ting the expression of survivin mRNA was exam-inedincervical cancer cell lines using molecular bea-coni maging technology.MATERIALS AND METHODS1Cervical cancer cell lines and ce...

  14. Influence of Ginkgo biloba extract on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma of lacrimal gland

    Institute of Scientific and Technical Information of China (English)

    Li-Xiao Zhou; Yu Zhu

    2012-01-01

    Objective: To explore the influence of extract of Ginkgo biloba (EGB) on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma (ACC) of lacrimal gland. Methods:ACC-2 cell in human with ACC of lacrimal gland was in vitro cultured. MTT method was used for cell proliferation detection. Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle. Survivin gene expression was analyzed by RT-PCR and Western blotting. Results: EGB had inhibitory effect on the proliferation of ACC-2 cell with significant dose-effect relationship, and there was statistical difference when compared with the control group (P<0.01). The inhibitory concentration 50 % (IC50) is 88 mg/L. The flow cytometer test indicated that EGB can gradually increase ACC-2 cell in G0-G1 stage and decrease it in G2-M and S stage. With the increase of dose, the apoptosis rate of ACC-2 cell was obviously increased (P<0.05 or P<0.01). EGB had certain inhibitory effect on Survivin gene expression of ACC-2 cell, and Survivin gene expression was decreased with the increasing of the EGB concentration (P<0.01). Conclusions:EGB can effectively inhibit Survivin gene expression of ACC-2 cell in human with ACC of lacrimal gland, induce the apoptosis of ACC-2 cell and inhibit tumor cell proliferation.

  15. Expression and biology significance of Survivin and VEGF in cervical carcinoma tissues%宫颈癌组织Survivin与 VEGF表达及其生物学意义的探讨

    Institute of Scientific and Technical Information of China (English)

    李淼; 钱智; 韩晓宇

    2011-01-01

    OBJECTIVE: To study the relationship between the expressions of Survivin and VEGF in cervical carcinoma and the clinical feature of cervical carcinoma, and to observe the correlation between Survivin and VEGF in cervical carcinoma. METHODS: Using streptavidin-biotin peroxidase (SP) method, the expressions of Survivin and VEGF were examined in 73 cases of cervical carcinoma, 25 cases of cervical intraepithelial neoplasia (CIN) and 20 cases of chronic cervicitis. RESULTS: Survivin and VEGF positive expression rate in cervical carcinoma group respectively were 75. 3% and 78. 1%, chronic cervical inflammation group were 5. 0% and 10. 0%, CIN group were 48. 0% and 52. 0%. The expression of Survivin and VEGF present a growing trend from chronic cervicitis to cervical carcinoma. The expression of Survivin and VEGF were related to the clinical stages and lymph node metastasis of cervical carcinoma. A positive correlation existed among Survivin and VEGF ( r = 0. 738, P<0. 01). CONCLUSION: The expressions of Survivin and VEGF in cervical carcinoma reflect the local invasion activity and metastasizing proficiency of cervical carcinoma, which are closely related to metastasis, clinical stages and prognosis of cervical carcinoma, and have synergistic effect in incidence and development of cervical carcinoma.%目的:探讨宫颈癌组织存活素(Survivin)与血管内皮生长因子(VEGF)的表达与宫颈癌临床特征的关系以及两者的相关性.方法:采用免疫组化SP法检测73例宫颈癌,25例宫颈上皮内瘤样病变(CIN)和20例慢性宫颈炎组织Survivin和VEGF的表达水平.结果:宫颈癌组织Survivin和VEGF阳性表达率分别为75.3%和78.1%,慢性宫颈炎组分别为5.0%和10.0%,CIN组分别为48.0%和52.0%.从慢性宫颈炎组到宫颈癌组,Survivin和VEGF表达率逐渐升高,P<0.05.Survivin和VEGF的表达与宫颈癌临床分期及淋巴结转移相关.Survivin表达与VEGF表达呈正相关,r=0.738,P<0.01.结论:Survivin

  16. Cyclin B1和survivin在非小细胞肺癌中的表达和意义%The expression and significance of cyclin B1 and survivin in human non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    Guosheng Li; Xuhan Liu; Daorong Zhang; Dong Liu; Zhiyong Li

    2011-01-01

    Objective: We studied the expression of cyclin B1 and survivin in human non-small cell lung cancer (NSCLC), and the relationship between such expression and clinicopathological features of NSCLC. Methods: One hundred cases of tissue specimen including NSCLC, neighboring noncancerous tissue and normal lung tissue were collected at random. These specimens were detected by immunohistochemical methods. Results: The expression of cyclin B1 and survivin showed significant difference (P 0.05) in NSCLC. Statistical significance was marked between different clinical stages of NSCLC and the expression of cyclin B1 and survivin (P < 0.05). Conclusion: The overexpression of cyclin B1 and survivin was found in NSCLC. The expression of cyclin B1 and survivin might be up-regulated during an early step of tumorigenesis and during the development of NSCLC. The progression of cell cycle could be efficiently connected with the control of apoptosis by the interrelations between the overexpression of cyclin B1 and that of survivin in NSCLC during the G2/M phase. The overexpression of cyclin B1 and survivin might be used as marker in showing the dividing and proliferating ability, and the inhibiting apoptosis ability (lengthening cell lifespan) of NSCLC. Moreover, the overexpression of cyclin B1 and survivin was associated with the clinic stages of NSCLC.

  17. Survivin knockdown increased anti-cancer effects of (-)-epigallocatechin-3-gallate in human malignant neuroblastoma SK-N-BE2 and SH-SY5Y cells

    Energy Technology Data Exchange (ETDEWEB)

    Hossain, Md. Motarab [Department of Pathology, Microbiology, and Immunology, University of South Carolina School of Medicine, Columbia, SC (United States); Banik, Naren L. [Department of Neurosciences, Medical University of South Carolina, Charleston, SC (United States); Ray, Swapan K., E-mail: swapan.ray@uscmed.sc.edu [Department of Pathology, Microbiology, and Immunology, University of South Carolina School of Medicine, Columbia, SC (United States)

    2012-08-01

    Neuroblastoma is a solid tumor that mostly occurs in children. Malignant neuroblastomas have poor prognosis because conventional chemotherapeutic agents are hardly effective. Survivin, which is highly expressed in some malignant neuroblastomas, plays a significant role in inhibiting differentiation and apoptosis and promoting cell proliferation, invasion, and angiogenesis. We examined consequences of survivin knockdown by survivin short hairpin RNA (shRNA) plasmid and then treatment with (-)-epigallocatechin-3-gallate (EGCG), a green tea flavonoid, in malignant neuroblastoma cells. Our Western blotting and laser scanning confocal immunofluorescence microscopy showed that survivin was highly expressed in malignant neuroblastoma SK-N-BE2 and SH-SY5Y cell lines and slightly in SK-N-DZ cell line. Expression of survivin was very faint in malignant neuroblastoma IMR32 cell line. We transfected SK-N-BE2 and SH-SY-5Y cells with survivin shRNA, treated with EGCG, and confirmed knockdown of survivin at mRNA and protein levels. Survivin knockdown induced morphological features of neuronal differentiation, as we observed following in situ methylene blue staining. Combination of survivin shRNA and EGCG promoted neuronal differentiation biochemically by increases in the expression of NFP, NSE, and e-cadherin and also decreases in the expression of Notch-1, ID2, hTERT, and PCNA. Our in situ Wright staining and Annexin V-FITC/PI staining showed that combination therapy was highly effective in inducing, respectively, morphological and biochemical features of apoptosis. Apoptosis occurred with activation of caspase-8 and cleavage of Bid to tBid, increase in Bax:Bcl-2 ratio, mitochondrial release of cytochrome c, and increases in the expression and activity of calpain and caspase-3. Combination therapy decreased migration of cells through matrigel and inhibited proliferative (p-Akt and NF-{kappa}B), invasive (MMP-2 and MMP-9), and angiogenic (VEGF and b-FGF) factors. Also, in vitro

  18. Expression and significance of survivin in rheumatoid arthritis synovium%存活素在类风湿关节炎滑膜组织中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    马莎; 林俊; 杨慧; 晋松; 李芹; 郑红梅; 王静; 杨永红

    2014-01-01

    目的 通过检测RA患者滑膜组织中存活素表达水平,分析其与RA患者临床特征之间的关系,并探讨存活素在RA发病机制中的作用.方法 用免疫组织化学的方法检测65例RA患者,9例OA患者,21份对照组滑膜组织中存活素表达水平,RA患者根据DAS28分为活动组与缓解组.记录入组患者临床资料:性别、年龄、病程、ESR、CRP、RF、CCP、DAS28评分.检测各组存活素蛋白的表达情况,并分析RA活动组存活素蛋白表达与其临床病理参数的相关性,多组间均数的比较使用单因素方差分析,率的比较采用Pearson x2检验或似然比x2检验,各指标与临床指标的相关性分析采用Spearman相关分析.结果 ①RA患者组滑膜组织中存活素表达水平明显高于OA组及正常对照组(P=0.001).②RA活动组患者滑膜组织中存活素表达水平明显高于缓解组(x2=41.62,P=0.001).③RA活动组患者存活素表达水平与ESR、CRP、DAS28评分呈正相关(r值分别为:0.297、0.411、0.416,P<0.05),而与CCP、RF、性别、年龄、病程无相关性(r值分别为:0.004、-0.114、-0.074、-0.0.35、-0.188,P>0.05).结论 存活素在RA滑膜组织中表达水平增高提示了其与滑膜成纤维细胞的凋亡异常相关,参与了RA的发病过程,检测存活素水平有助于对RA疾病活动性的评估.%Objective To investigate the role of survivin in the pathogenesis of rheumatoid arthritis (RA) by detecting of expression levels of survivin in synovial tissue of patients with rheumatoid arthritis and analyze its relationship with clinical features of RA patients.Methods The expression levels of survivin in synovial tissue of patients with RA were detected by immunohistochemistry method in 65 cases of RA,9 cases of osteoarthritis,21 normal controls.According to the disease activity score (DAS28),RA patients were divided into active disease group and remission group.Clinical data of these patients were recorded,including gender

  19. Survivin mRNA expression in urine as a biomarker for patients with transitional cell carcinoma of bladder

    Institute of Scientific and Technical Information of China (English)

    HOU Jian-quan; HE Jun; WEN Duan-gai; CHEN Zi-xing; ZENG Jian

    2006-01-01

    @@ Transitional cell carcinoma (TCC) of bladder is the most common malignant tumor in uropoiesis system. Up to date, there is still lack of an ideal marker for the diagnosis of TCC except CT and MRI imaging and cystoscopy. Cystoscopy is an invasive examination, which increases the possibility of urinary tract infection. Urine cytology has low sensitivity (21%-40%) in diagnosis of bladder cancer, especially for those with medium or high differentiation. The specificity is often affected by factors such as specimen collection, urinary tract infection, etc. Detecting the expression of survivin mRNA in urine by real time-PCR is simple in specimen collection and is sensitive and relatively specific, which provides a simple and noninvasive diagnostic method for TCC. Moreover it allows comparing the gene expression levels at different stages and grades of TCC, which can help define malignancy degree of TCC.

  20. Increased NQO1 but Not c-MET and Survivin Expression in Non-Small Cell Lung Carcinoma with KRAS Mutations

    Directory of Open Access Journals (Sweden)

    Ahmet Yilmaz

    2014-09-01

    Full Text Available Cigarette smoking is one of the most significant public health issues and the most common environmental cause of preventable cancer deaths worldwide. EGFR (Epidermal Growth Factor Receptor-targeted therapy has been used in the treatment of LC (lung cancer, mainly caused by the carcinogens in cigarette smoke, with variable success. Presence of mutations in the KRAS (Kirsten rat sarcoma viral oncogene homolog driver oncogene may confer worse prognosis and resistance to treatment for reasons not fully understood. NQO1 (NAD(PH:quinone oxidoreductase, also known as DT-diaphorase, is a major regulator of oxidative stress and activator of mitomycins, compounds that have been targeted in over 600 pre-clinical trials for treatment of LC. We sequenced KRAS and investigated expression of NQO1 and five clinically relevant proteins (DNMT1, DNMT3a, ERK1/2, c-MET, and survivin in 108 patients with non-small cell lung carcinoma (NSCLC. NQO1, ERK1/2, DNMT1, and DNMT3a but not c-MET and survivin expression was significantly more frequent in patients with KRAS mutations than those without, suggesting the following: (1 oxidative stress may play an important role in the pathogenesis, worse prognosis, and resistance to treatment reported in NSCLC patients with KRAS mutations, (2 selecting patients based on their KRAS mutational status for future clinical trials may increase success rate, and (3 since oxidation of nucleotides also specifically induces transversion mutations, the high rate of KRAS transversions in lung cancer patients may partly be due to the increased oxidative stress in addition to the known carcinogens in cigarette smoke.

  1. BRMS1和 Survivin 在乳腺癌中的表达及临床意义%Expression and clinical significance of BRMS1 and Survivin in breast cancer

    Institute of Scientific and Technical Information of China (English)

    王大勇

    2014-01-01

    Objective To investigate the expression and clinical significance of BRMS1 and Survivin in breast cancer,and to explore the correlation between them and their effects on the pathogenesis ,development,metastasis and prognosis of breast cancer.Methods The expression levels of BRMS1 and Survivin were detected by immunohistochemistry in 160 cases of breast cancer,70 cases of mammary glands benign proliferative lesions and 40 cases of normal tissues adjacent to breast cancer.The data was analyzed together with clinical pathological parameters .Results The positive expression rates of BRMS1in breast cancer, mammary glands benign proliferative lesions and normal tissues adjacent to breast cancer were 34.4%,81.4%,95.0%,respectively,the positive expression rates of BRMS1 in breast cancer were significantly decreased ,as compared with those in mammary glands benign proliferative lesions and normal tissues adjacent to breast cancer ( P <0.05).However the positive expression rates of Survivin in breast cancer ,mammary glands benign proliferative lesions and normal tissues adjacent to breast cancer were 78.8%,17.1%,0,respectively,the positive expression rates of Survivin breast cancer were significantly increased,as compared with those in mammary glands benign proliferative lesions and normal tissues adjacent to breast cancer ( P <0.05).The abnormal expression of BRMS1 and Survivin in breast cancer tissue was correlated to TNM clinical stage and lymph node metastasis .Survivin was also related to histological grade .The abnormal expression of BRMS1 and Survivin in breast cancer tissue was not related to patient's age and tumor's size,but the expression of BRMS1 was negatively correlated with that of Survivin .Conclusion BRMS1 and Survivin play an important role in the carcinogenesis , development,metastasis and prognosis of breast cancer ,which can be used as the markers in diagnosis and prognosis evaluation of breast cancer.%目的:观察BRMS1和Survivin在人乳腺癌组

  2. Inhibition of serine-peptidase activity enhances the generation of a survivin-derived HLA-A2-presented CTL epitope in colon-carcinoma cells.

    Science.gov (United States)

    Preta, G; Marescotti, D; Fortini, C; Carcoforo, P; Castelli, C; Masucci, M; Gavioli, R

    2008-12-01

    Cytotoxic T lymphocytes eliminate tumor cells expressing antigenic peptides in the context of MHC-I molecules. Peptides are generated during protein degradation by the proteasome and resulting products, surviving cytosolic amino-peptidases activity, may be presented by MHC-I molecules. The MHC-I processing pathway is altered in a large number of malignancies and modulation of antigen generation is one strategy employed by cells to evade immune control. In this study we analyzed the generation and presentation of a survivin-derived CTL epitope in HLA-A2-positive colon-carcinoma cells. Although all cell lines expressed the anti-apoptotic protein survivin, some tumors were poorly recognized by ELTLGEFLKL (ELT)-specific CTL cultures. The expression of MHC-I or TAP molecules was similar in all cell lines suggesting that tumors not recognized by CTLs may present defects in the generation of the ELT-epitope which could be due either to lack of generation or to subsequent degradation of the epitope. The cells were analyzed for the expression and the activity of extra-proteasomal peptidases. A significant overexpression and higher activity of TPPII was observed in colon-carcinoma cells which are not killed by ELT-specific CTLs, suggesting a possible role of TPPII in the degradation of the ELT-epitope. To confirm the role of TPPII in the degradation of the ELT-peptide, we showed that treatment of colon-carcinoma cells with a TPPII inhibitor resulted in a dose-dependent increased sensitivity to ELT-specific CTLs. These results suggest that TPPII is involved in degradation of the ELT-peptide, and its overexpression may contribute to the immune escape of colon-carcinoma cells.

  3. 天花粉蛋白对子宫颈癌Hela田胞Survivi基因的影响%Effects of trichosanthin on expression of Survivin gene in Hela cell line

    Institute of Scientific and Technical Information of China (English)

    谭寒星; 黄利鸣; 王艳林; 韩钰; 刘朝奇; 尤程程; 黄玉蓉

    2011-01-01

    目的:研究天花粉蛋白(TCS)对子宫颈癌Hela细胞Survivin基因表达的影响,以探讨TCS诱导Hela细胞凋亡的机制.方法:MTT法检测TCS对Hela细胞增殖的影响,倒置显微镜观察细胞形态;RT-PCR和Western blot 分析TCS对Hela细胞Survivin基因表达的影响;流式细胞仪分析TCS对Hela细胞周期的影响.结果:①TCS显著性抑制Hela细胞增殖;②rCS处理后Hela细胞Survivin mRNA水平无明显改变,但蛋白表达水平显著降低;③YCS降低Hela细胞周期中G2/M期细胞比率,但增加S期细胞的数量.结论:通过下调凋亡抑制因子Survivin的表达而诱发Hela细胞凋亡,可能是TCS抑制Hela细胞增殖的分子机制.%Objective: To observe the effects of Trichosanthin (TCS) on expression of Survivin gene in Hela cell line and to explore the underlying molecular mechanism on the growth inhibition and the apoptosis of Hela cell line induced by TCS. Methods: MTT assay was used to determine the effect of TCS on cell proliferation; the cell morphology was observed by inverted microscope. RT-PCR and Western-blot assay were used to detect expression level of Survivin gene in Hela cells after TCS treatment. The FCM analysis was carried out to examine the effect of TCS on the cell cycle of Hela cells. Results: ①The proliferation of Hela cells was significantly inhibited by TCS; ②TCS treatment decreased expression of Survivin in the protein level, but no change was observed in the mRNA level; ③TCS treatment resulted in deceased cell number in G2/M phase but increased cell number in S phase. Conclusion: TCS inhibits the growth of Hela cells through decreasing the expression of Survivin, which results in activation of apoptosis pathways.

  4. Mitotic slippage and expression of survivin are linked to differential sensitivity of human cancer cell-lines to the Kinesin-5 inhibitor monastrol.

    Directory of Open Access Journals (Sweden)

    Hila Asraf

    Full Text Available The mitotic Kinesin-5 motor proteins crosslink and slide apart antiparallel spindle microtubules, thus performing essential functions in mitotic spindle dynamics. Specific inhibition of their function by monastrol-like small molecules has been examined in clinical trials as anticancer treatment, with only partial success. Thus, strategies that improve the efficiency of monastrol-like anticancer drugs are required. In the current study, we examined the link between sensitivity to monastrol and occurrence of mitotic slippage in several human cell-lines. We found that the rank of sensitivity to monastrol, from most sensitive to least sensitive, is: AGS > HepG2 > Lovo > Du145 ≥ HT29. We show correlation between the sensitivity of a particular cell-line to monastrol and the tendency of the same cell-line to undergo mitotic slippage. We also found that in the monastrol resistant HT29 cells, prolonged monastrol treatments increase mRNA and protein levels of the chromosomal passenger protein survivin. In contrast, survivin levels are not increased by this treatment in the monastrol-sensitive AGS cells. We further show that over-expression of survivin in the monastrol-sensitive AGS cells reduces mitotic slippage and increases resistance to monastrol. Finally, we show that during short exposure to monastrol, Si RNA silencing of survivin expression reduces cell viability in both AGS and HT29 cells. Our data suggest that the efficiency of anti-cancer treatment with specific kinesin-5 inhibitors may be improved by modulation of expression levels of survivin.

  5. Plasmid-based Survivin shRNA and GRIM-19 carried by attenuated Salmonella suppresses tumor cell growth

    Institute of Scientific and Technical Information of China (English)

    Yan-Bo Liu1; De-Qi Xu; Ling Zhang1; Ya-Xiong Guo; Li-Fang Gao; Xi-Chun Liu; Li-Juan Zhao; Bao-Feng Guo; Li-Jing Zhao; Xue-Jian Zhao

    2012-01-01

    Persistent activation of Survivin and its overexpression contribute to the formation,progression and metastasis of several different tumor types.Therefore,Survivin is an ideal target for RNA interference mediated-growth inhibition.Blockade of Survivin using specific short hairpin RNAs (shRNA) can significantly reduce prostate tumor growth.RNA interference does not fully ablate target gene expression,owing to the idiosyncrasies associated with shRNAs and their targets.To enhance the therapeutic efficacy of Survivin-specific shRNA,we employed a combinatorial expression of Survivin-specific shRNA and gene associated with retinoid-interferon-induced mortality-19 (GRIM-19).Then,the GRIM-19 coding sequences and Survivin-specific shRNAs were used to create a dual expression plasmid vector and were carried by an attenuated strain of Salmonella enteric serovar typhimurium (S.typhimurium) to treat prostate cancer in vitroand in vivo.We found that the co-expressed Survivin-specific shRNA and GRIM-19synergistically and more effectively inhibited prostate tumor proliferation and survival,when compared with treatment with either single agent alone in vitro and in vivo.This study has provided a novel cancer gene therapeutic approach for prostate cancer.

  6. MGMT和Survivin在乳腺癌中的表达及其临床意义%Expression of MGMT and Survivin in Breast Cancer and Its Clinical Significance

    Institute of Scientific and Technical Information of China (English)

    王发亮; 薄爱华; 芦广萍; 侯金超; 吴荣薇

    2008-01-01

    Objective:To investigate the expression of MGMT and Survivin in breast cancer and its clinical significance.Methods:Specimens from breast adenoma and breast cancer were taken and had been Formalin-fixed,paraffin-embedded.With Strept Avidin-Biotin Complex(SABC) immunohistochemical technique,the expression of MGMT and Survivin in these specimens was examined.Results:It was found that there were significant differences between MGMT and Survivin in breast adenoma and breast lymph node metastasis,while the expression of Survivin was associated with lymph node metastasis only.Meanwhile,the expression of MGMT was correlated with that of Survivin(r=0.34,P<0.01).Conclusion:It was concluded that the abnormal expressions of MGMT and Survivin were associated with lymph node metastasis.They possibly could be useful indexes for the primary screening and the prognosis of breast cancer.The examination of them may have an important guiding significance in the chemotherapy strategy.%目的:研究MGMT和Survivin在乳腺癌中的表达及其临床意义.方法:福尔马林固定,石蜡包埋乳腺癌和腺瘤标本,采用SABC免疫组织化学方法检测MGMT和Survivin在这些组织中的表达.结果:MGMT和Survivin在乳腺癌和乳腺腺瘤中的表达有显著性差异.MGMT在乳腺癌中的表达与患者的年龄、淋巴结转移有关,而Survivin仅与淋巴结转移有关.另外,MGMT和Survivin之间具有相关性(r=0.48,P<0.01).结论:MGMT和Survivin的异常表达与乳腺癌的淋巴结转移有关;MGMT和Survivin可以作为判断乳腺癌发生和预后的重要指标;检测它们的表达可以指导临床上化疗方案的制定.

  7. PTEN与Survivin在人皮肤血管瘤组织中的表达及意义%Significance and Expression of PTEN and Survivin in Human Dermal Hemangioma

    Institute of Scientific and Technical Information of China (English)

    蒋晖; 汪晓庆; 张端莲; 陕声国; 吴慧芬; 蔡丽华; 袁玉虎

    2009-01-01

    目的 探讨PTEN与Survivin在血管瘤发生、发展过程中的表达及意义.方法 应用免疫组织化学方法和RT-PCR法检测了皮肤血管瘤增生期、退化期以及正常皮肤组织中PTEN和Survivin的表达水平.结果 ①免疫组织化学结果:PTEN在增生期血管瘤内皮细胞的表达低于退化期,差异有显著性意义(P0.05).②RT-PCR结果:在退化期血管瘤和正常皮肤组织中均有明显的PTEN mRNA表达,而增生期血管瘤中PTEN mRNA的表达较弱;在增生期血管瘤中有明显的Survivin mRNA表达,而退化期血管瘤和正常皮肤组织中Survivin mRNA均无表达.结论 PTEN和Survivin参与了血管瘤的发生、发展和退化,PTEN通过诱导内皮细胞凋亡而促进血管瘤由增生向退化的转变,Survivin通过抑制内皮细胞凋亡而促进血管瘤的增生,两者在血管瘤的发生发展中发挥协同效应.%Objective To study the expression of PTEN and Survivin,and investigate the mechanism and significance of them in different phases of human hemangiomas. Methods The expression of PTEN and Survivin was detected by using immu-no-histochemical technique,and that of PTEN mRNA and Survivin mRNA by using reverse transcription polymerase chain reac-tion(RT-PCR)in proliferating,involuting human hemangiomas and normal skin tissues. Results ①The expression of PTEN in the endothelial cells of involuting hemangiomas was significantly higher than in proliferating hemangiomas(P0. 05). ②The expression of PTEN mRNA was strong in involting hemangiomas and normal skin tissues, but weak in proliferating hemangiomas. The expression of Survivin mRNA was significantly increased in proliferating hemangiomas,and no Survivin mRNA was detected in involuting hemangiomas and normal skin tissues. Conclusion It is suggested that both PTEN and Survivin may take part in the genesis,development,and involution. PTEN promoted the switching from proliferation to involution in hemangiomas through inducing the

  8. Lupeol triterpene, a novel diet-based microtubule targeting agent: disrupts survivin/cFLIP activation in prostate cancer cells.

    Science.gov (United States)

    Saleem, Mohammad; Murtaza, Imtiyaz; Witkowsky, Olya; Kohl, Amanda Marie; Maddodi, Nityanand

    2009-10-23

    Recently we showed Lupeol, a triterpene, found in fruits and vegetables inhibits the growth of tumors originated from human androgen-sensitive prostate cancer (CaP) cells and decreases the serum-PSA levels in a mouse model. Here, we provide evidence that Lupeol inhibits the growth of androgen-sensitive as well as androgen-insensitive CaP cells by inducing G2/M cell cycle arrest without exhibiting any toxicity to normal human prostate epithelial cells (PrEC) at the doses at which it kills cancer cells. We observed that Lupeol treatment to LNCaP and DU145 cells resulted in a dose-dependent (i) decrease in the protein levels of Cyclins-A, -B1, -D1, -D2, -E2, cyclin-dependent kinase (cdk)-2 and (ii) increase in the protein level of CDK-inhibitor p21. Since G2/M cell cycle phase is regulated by microtubule assembly, we investigated effect of Lupeol on microtubule assembly, its regulation and down-stream targets in CaP cells. Lupeol treatment significantly modulated the level of (i) microtubule components alpha-tubulin and beta-tubulin, (ii) microtubule-regulatory protein stathmin, and (iii) microtubule-regulatory down-stream target/pro-survival protein survivin. Lupeol treatment also decreased the level of anti-apoptotic protein cFLIP. Finally, Lupeol was observed to significantly decrease the transcriptional activation of survivin and cFLIP genes in CaP cells. We conclude that the Lupeol-induced growth inhibition of CaP cells is a net outcome of simultaneous effects on stathmin, cFLIP, and survivin which results in the disruption of microtubule assembly. We suggest that Lupeol alone or as an adjuvant to other microtubule agents could be developed as a potential agent for the treatment of human CaP.

  9. P120ctn overexpression enhances β-catenin-E-cadherin binding and down regulates expression of survivin and cyclin D1 in BEL-7404 hepatoma cells

    Institute of Scientific and Technical Information of China (English)

    Chao-Zan Nong; Li-Li Pan; Wei-Sheng He; Xi-Liang Zha; Hai-Hong Ye; Hua-Yi Huang

    2006-01-01

    AIM: To understand the role of P120ctn in E-cadherin-mediated cell-cell adhesion and signaling as well as in hepatoma cell biological function.METHODS: We stably overexpressed p120ctn isoform 3A in BEL-7404 human hepatoma cells and studied the effect of p120ctn on β-catenin and E-cadherin binding as well as p120ctn and β-catenin subcellular localization using immunoprecipitation, Western blotting and confocalmicroscopy. We also investigated the inhibitory effect of p120ctn transfection on the expression of apoptotic protein survivin survivin and cell cycle regulator cyclin D1in the cells.RERULTS: Western blotting indicated that p120ctn expression increased after cells were transfected with p120ctn isoform 3A. The protein was located mainly at membrane under immunofluorescent microscope.β-catenin nuclear expression was reduced after overexpression of p120ctn isoform 3A. The p120ctn-E-cadherin binding increased after transfection of p120ctn isoform 3A. Furthermore, overexpression of p120ctn down regulated the expression of apoptotic protein survivin and cell cycle regulator cyclin D1. These effects led to reduction of cell proliferation.CONCLUSION: Our results indicate that p120ctn plays an important role in regulating the formation of E-cadherin and -catenin complex, cell apoptosis, cell cycle and cancer cell biological function.

  10. Preclinical development of HIvax: Human survivin highly immunogenic vaccines.

    Science.gov (United States)

    Hoffmann, Peter R; Panigada, Maddalena; Soprana, Elisa; Terry, Frances; Bandar, Ivo Sah; Napolitano, Andrea; Rose, Aaron H; Hoffmann, Fukun W; Ndhlovu, Lishomwa C; Belcaid, Mahdi; Moise, Lenny; De Groot, Anne S; Carbone, Michele; Gaudino, Giovanni; Matsui, Takashi; Siccardi, Antonio; Bertino, Pietro

    2015-01-01

    Our previous work involved the development of a recombinant fowlpox virus encoding survivin (FP-surv) vaccine that was evaluated for efficacy in mesothelioma mouse models. Results showed that FP-surv vaccination generated significant immune responses, which led to delayed tumor growth and improved animal survival. We have extended those previous findings in the current study, which involves the pre-clinical development of an optimized version of FP-surv designed for human immunization (HIvax). Survivin-derived peptides for the most common haplotypes in the human population were identified and their immunogenicity confirmed in co-culture experiments using dendritic cells and T cells isolated from healthy donors. Peptides confirmed to induce CD8(+) and CD4(+) T cells activation in humans were then included in 2 transgenes optimized for presentation of processed peptides on MHC-I (HIvax1) and MHC-II (HIvax2). Fowlpox vectors expressing the HIvax transgenes were then generated and their efficacy was evaluated with subsequent co-culture experiments to measure interferon-γ and granzyme B secretion. In these experiments, both antigen specific CD4(+) and CD8(+) T cells were activated by HIvax vaccines with resultant cytotoxic activity against survivin-overexpressing mesothelioma cancer cells. These results provide a rationale for clinical testing of HIvax1 and HIvax2 vaccines in patients with survivin-expressing cancers.

  11. Survivin和Smac在卵巢黏液性肿瘤中的表达及意义%Expression and significance of Survivin and Smac in ovarian mutinous tumors

    Institute of Scientific and Technical Information of China (English)

    王红霞; 陈钢; 李国利; 江亚军

    2010-01-01

    Objective To investigate the expressions and significances of Survivin and Smac in ovarian mucinous tumors. Methods A total of 55 paraffin-embedded specimens of primary ovarian mucinous tumors were collected. SABC was used to detect protein expression of Survivin and Smac genes.Immunoelectron microscopy using colloidal gold labeling was performed to determine the subcellular localization and patterns of Smac protein expression. Results (1) The cytoplasmic expression rates of survivin in benign, borderline and malignant ovarian mucinous tumors were 2/20, 12/15 and 20/20 respectively, which presents an improving trend. There were significant differences of survivin expression between benign vs. Borderline lesions (P 0. 05). Conclusions With the malignant development of ovarian mucinous tumors,the expressions of Survivin are up-regulated,and the expressions of Smac are down-regulated. Smac proteins exist mainly in an inactive intramembranous storage form inside of mitochondria.%目的 探讨Survivin和Smae在卵巢黏液性肿瘤中的表达及意义.方法 应用免疫组织化学SABC法检测55例原发性卵巢黏液性肿瘤中的Survivin和Smac表达,并应用免疫电镜胶体金标记法观察Smac在原发性卵巢黏液性肿瘤中的亚细胞定位及表达趋势.结果 (1)Survivin在良性、交界性和恶性组的阳性表达率分别为2/20、12/15和20/20,呈升高趋势,其中良性与交界性、良性与恶性组间比较差异均有统计学意义(P0.05).结论 随着卵巢黏液性肿瘤的恶性进展,凋亡抑制蛋白Survivin表达上调,促凋亡蛋白Smac表达下调;下调的Smac蛋白表达主要为储存在线粒体膜间的非活性形式蛋白,而非释放到胞质中的活性形式蛋白.

  12. Survivin在鼻息肉组织中的表达及意义%The Expression and Significance of Survivin in Nasal Polyps

    Institute of Scientific and Technical Information of China (English)

    王巍毅; 肖旭平; 冯晓辉

    2011-01-01

    [目的]探讨凋亡抑制蛋白survivin在鼻息肉组织中的表达及意义.[方法]用免疫组化染色法检测鼻息肉组(40例)、钩突炎症黏膜组(20例)和正常下甲黏膜组(20例)中survivin的表达.[结果]①survivin在几乎所有的炎症细胞中均呈阳性表达,三组间无明显差异.②观察survivin在黏膜上皮和腺上皮中的表达,显示鼻息肉组的阳性表达明显高于其他两组,差异有显著性(P<0.01),而其他两组之间的阳性表达差异无显著性(P>0.05).[结论]survivin在正常鼻黏膜表达可能对维持鼻腔的结构功能具有一定的生物学意义,并且它介导的凋亡抑制现象对鼻息肉的发生发展可能也具有一定的作用.%[Objcctive]To investigate the expression of apoptosis inhibitor protein survivin in nasal polyps and explore its role in the pathogencsis of nasal polyps.[Methods]The expression of survivin was determined in nasal polyps group( n =40) , uncinate process mucous inflammation group( n =20) and normal inferior tur binate mucous group( n =20) by immunohistochemical staining.[Results]The positive expression of survivin was found in almost all inflammatory cells, and there was no obvious difference among 3 groups.The positive expression in epithelium mucosae and glandular epithelium in nasal polyps group were stronger than that in other two groups, and there was significant difference between nasal polyps group and other two groups( P < 0.01) , but there was no significant difference between other two groups( P >0.05).[Conclusion]The expression of survivin in normal nasal mucosa may have biological significance for maintaining the structure and function of nasal cavity.Survivin-mediated apoptosis inhibition may play an important role in the pathogencsis and development of nasal polyps.

  13. Berberine and Curcumin Target Survivin and STAT3 in Gastric Cancer Cells and Synergize Actions of Standard Chemotherapeutic 5-Fluorouracil.

    Science.gov (United States)

    Pandey, Arvind; Vishnoi, Kanchan; Mahata, Sutapa; Tripathi, Satyendra Chandra; Misra, Sri Prakash; Misra, Vatsala; Mehrotra, Ravi; Dwivedi, Manisha; Bharti, Alok C

    2015-01-01

    Aberrantly expressed survivin and STAT3 signaling have emerged as major determinants of chemoresistance in gastric cancer. We evaluated effects of potent herbal derivatives curcumin, berberine, and quercetin on STAT3 signaling, survivin expression, and response to 5-fluorouracil (5-FU) treatment in gastric cancer cells (AGS). Cytotoxic and inhibitory effects of berberine, curcumin, and quercetin alone or in combination with 5-FU were examined by MTT assay, and their effect on survivin, STAT3, and the phosphorylated active STAT3 (pSTAT3) expression was examined by western blotting. Effect of these herbal derivatives on STAT3 DNA binding activity was measured by electrophoretic mobility shift assay. Curcumin, berberine, and quercetin effectively downregulated pSTAT3 levels, survivin expression, and gastric cancer cells viability in a dose-dependent manner (with corresponding IC50 values of 40.3μM, 29.2μM and 37.5μM, respectively). Berberine was more effective in inhibiting survivin expression as compared to other herbal agents. 5-FU in combination with berberine or curcumin showed a synergistic inhibition of survivin and STAT3 level resulting in enhanced cell death in gastric cancer cells. Overall, our data suggest use of berberine and curcumin as adjunct therapeutics to overcome chemoresistance during treatment of gastric malignancies.

  14. 卵巢上皮性肿瘤中PIEN和Survivin的表达及意义%Expression and significance of SUrvivin and PIEN in epithelial ovarian tumors

    Institute of Scientific and Technical Information of China (English)

    丁月红

    2011-01-01

    Objective To investigate the expression and significance of Survivin and PIEN in epithelial ovarian tumors.Methods Immunohistochemical detection of PTEN and Survivin in 52 cases of epithelial ovarian cancer, 20 cases of benign ovarian epithelial tumors and 18 normal ovarian tissue in rats.Results In normal ovarian tissues Survivin protein expression in non- benign epithelial ovarian tumors and epithelial ovarian cancer in the Survivin protein expression rates were 26.32% ( 5/19 ), 66.67% (30/45), epithelial ovarian cancer in the Survivin protein expression was significantly higher than that of normal ovarian and benign epithelial ovarian tumor group, the difference was significant ( P < 0.05 ); PTEN protein in normal ovarian tissue, benign epithelial ovarian tumors and epithelial ovarian carcinoma were 100.0% (18/18), 95.0% (20/19), 38.5% (20/52).PTEN protein in normal ovarian tissues and benign epithelial ovarian expression of swelling was higher than that of epithelial ovarian cancer (all P < 0.01 ).Conclusions PTEN expression was decreased and increased expression of Survivin and related to the occurrence of epithelial ovarian cancer, the detection of ovarian cancer patients Survivin and PIEN expression for the evaluation of malignancy, and prognosis has important clinical value.%目的 探讨卵巢上皮性肿瘤中PIEN和Survivin的表达及意义.方法 采用免疫组织化学法检测PTEN和survivin在52例卵巢上皮性癌、20例卵巢良性上皮性肿瘤和18例正常卵巢组织标本中的表达.结果 正常卵巢组织中无Survivin蛋白表达,良性上皮性卵巢肿瘤和卵巢上皮性癌中Survivin蛋白表达率分别为26.32%(5/19)、66.67%(30/45),卵巢上皮性癌中Survivin蛋白阳性表达率明显高于正常卵巢及良性上皮性卵巢肿瘤组,差异有显著性(P<0.05);PTEN蛋白在正常卵巢组织、卵巢良性上皮性肿瘤和卵巢上皮性癌中的表达率分别为100.0%(18/18)、95.0%(20/19)、38

  15. Theranostic properties of a survivin-directed molecular beacon in human melanoma cells.

    Directory of Open Access Journals (Sweden)

    Sara Carpi

    Full Text Available Survivin is an inhibitor of apoptosis overexpressed in different types of tumors and undetectable in most terminally differentiated normal tissues. In the current study, we sought to evaluate the in vitro theranostic properties of a molecular beacon-oligodeoxynucleotide (MB that targets survivin mRNA. We used laser scanning confocal microscopy to study MB delivery in living cells and real-time PCR and western blot to assess selective survivin-targeting in human malignant melanoma cells. We further assess the pro-apoptotic effect of MB by measuring internucleosomal DNA fragmentation, dissipation of mitochondrial membrane potential (MMP and changes in nuclear morphology. Transfection of MB into A375 and 501 Mel cells generated high signal intensity from the cytoplasm, while no signal was detected in the extracellular environment and in survivin-negative cells (i.e., human melanocytes and monocytes. MB time dependently decreased survivin mRNA and protein expression in melanoma cells with the maximum effect reached at 72 h. Treatment of melanoma cells with MB induced apoptosis by significant changes in MMP, accumulation of histone-complexed DNA fragments in the cytoplasm and nuclear condensation. MB also enhanced the pro-apoptotic effect of standard chemotherapeutic drugs tested at clinically relevant concentrations. The MB tested in the current study conjugates the ability of imaging with the pharmacological silencing activity against survivin mRNA in human melanoma cells and may represent an innovative approach for cancer diagnosis and treatment.

  16. 寻常型银屑病皮损中Stat3和Survivin的表达及相关性分析%The expression of Stat3 and Survivin in the lesions of psoriasis vulgaris and their correlation

    Institute of Scientific and Technical Information of China (English)

    冯世军; 高顺强; 李华蕊; 马敬; 李萍

    2012-01-01

    Objective To investigate the expression of Stat3 and Survivin in the lesions of psoriasis vulgaris and their correlation. Methods The expressions of Stat3 and Survivin in 52 patients with psoriasis vulgaris were detected by immunohistochemistry, and the correlation between Stat3 and Survivin expression was analyzed. Results The expressions of Stat3 and Survivin were significantly up-regulated, as compared with those in control group( P < 0. 05 ). The expression of Stat3 was positively correlated with that of Survivin in patients with psoriasis vulgaris ( P < 0. 01 ). Conclusion The Stat3 and Survivin are over-expressed in psoriasis vulgaris. Survivin can promote the proliferation of keratinocyte in the lesions of psoriasis vulgaris by up-regulating the expression of Survivin, which may play an important role in the pathogenesis and development of psoriasis vulgaris.%目的 探讨信号传导及转录活化因子3( Stat3)及生存素(Survivin)在寻常型银屑病皮损中的表达及其相关性.方法 通过免疫组化方法研究52例寻常型银屑病皮损中Stat3及Survivin表达的情况,并探讨Stat3和Survivin表达的相关性.结果 寻常型银屑病皮损中Stat3的表达和凋亡抑制蛋白Survivin的表达与正常对照组相比均明显上调(P<0.05).银屑病患者Stat3表达和Survivin表达呈正相关(P<0.01).结论 寻常型银屑病皮损中存在Stat3和Survivin的高表达,Stat3可能通过上调Survivin的表达,促进银屑病皮损处角质形成细胞的增殖,在寻常型银屑病的发生发展中起重要作用.

  17. Decrease of survivin, p53 and Bcl-2 expression in chemorefractory colorectal liver metastases may be predictive of radiosensivity radiosensivity after radioembolization with yttrium-90 resin microspheres.

    Science.gov (United States)

    Melucci, Elisa; Cosimelli, Maurizio; Carpanese, Livio; Pizzi, Giuseppe; Izzo, Francesco; Fiore, Francesco; Golfieri, Rita; Giampalma, Emanuela; Sperduti, Isabella; Ercolani, Cristiana; Sciuto, Rosa; Mancini, Raffaello; Garufi, Carlo; Diodoro, Maria Grazia; Mottolese, Marcella

    2013-03-06

    In a prospective multicenter phase II trial of radioembolization with yttrium-90 ((90)Y-RE) in chemorefractory liver-dominant metastatic colorectal cancer (mCRC), we showed that median survival was 12.6 months (95% CI 7.0-18.3) with 48% of 50 patients achieving disease control. In this extension retrospective study, we analyzed whether a panel of biomarkers, known to be associated to an adverse clinical outcome, underwent variations in CRC liver metastases pre and post (90)Y-RE.Of the 50 patients included in the study, 29 pre-(90)Y-RE therapy and 15 post-(90)Y-RE had liver biopsy specimens available. In these series we investigated survivin, p53, Bcl-2 and Ki-67 expression pre- and post-(90)Y-RE by immuhistochemistry (IHC). Our findings evidenced a decrease of survivin (77% vs 33%), p53 (93% vs 73%), Bcl-2 (37% vs 26%) expression as well as of Ki-67 proliferation index (62.5% vs 40%) on liver biopsies collected post-(90)Y-RE as compared to pre-(90)Y-RE. In the subset of 13 matched liver metastases we further confirmed the reduction of survivin (92.3% vs 53.8%; p = 0.06), p53 (100% vs 69.2%; p = 0.05) and Bcl-2 (69.2% vs 53.8%; p = 0.05) expression post-(90)Y-RE. This biomarker modulation was accompanied by morphological changes as steatohepatitis, hepatocyte necrosis, collagen deposition, proliferating and/or bile duct ectasia, focal sinusoidal dilatation and fibrosis.Although our analysis was conducted in a very limited number cases, these changes appear strictly related to the response to (90)Y-RE therapy and may deserve further investigation on a larger series of patients.

  18. 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.

  19. Expression of Survivin and Smac in thyroid tumors and its clinical significance%Survivin和Smac在甲状腺肿瘤中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    徐冬香; 游庆华; 王爱华; 叶惠英

    2011-01-01

    目的:观察Survivin和Smae在甲状腺肿瘤中的表达及探讨其意义.方法:采用免疫组织化学Envision法检测80例甲状腺肿瘤组织中的Survivin和Smac表达,按照阳性细胞数5%、25%、50%划分,是否出现表达,分析Survivin 表达与临床病理因素和Smac的关系.结果:Survivin在甲状腺腺瘤、甲状腺癌和伴有淋巴结转移的阳性表达率分别为20.0%(3/15),61.5%(40/65),80.0%(16/20),良性肿瘤和恶性肿瘤、良性肿瘤和恶性肿瘤伴有淋巴结转移两组间两两比较差异均有统计学意义(均P<0.05).Smac在甲状腺腺瘤和甲状腺癌和伴有淋巴结转移的阳性表达率分别为86.7%(13/15)、38.5%(25/65)、40.0%(8/20),良性肿瘤和恶性肿瘤、良性肿瘤和恶性肿瘤伴有淋巴结转移两组间两两比较差异有统计学意义(P<0.05).Survivin和Smac之间呈负相关(P<0.05).Survivin的表达与甲状腺癌患者的性别、肿瘤大小和肿瘤病理类型无关(R2=0.770 2,R2=0.632 5,R2=0.750 0),与年龄、TNM分期和淋巴结转移明显相关,差异有统计学意义(R2=0.986 5,R2=0.931 1,R2=0.964 3).Smac表达与年龄及淋巴结转移明显相关(R2=0.993 3,R2=0.925 9),而与性别、肿瘤大小、病理类型、TNN分期无关(R2=0.714 7,R2=0.793 7,R2=0.831 2,R2=0.816 3).结论:Survivin随着甲状腺肿瘤的恶性程度增加表达上调,而Smac表达下调,提示Survivin和Smac在甲状腺肿瘤诊断和鉴别诊断中具有一定的价值.%Objective: To observe expression of Survivin and Smac in thyroid tumors and explore its significance.Methods: Envision immunohistochernistry detected 80 cases of thyroid tumors in the expression of Survivin and Smac, in accordance with 5% positive cells, 25%, 50% of the division, whether there was expression, relationship between expression of Survivin and clinicopathologic factors and Smac were analysed.Results: Survivin in thyroid adenoma, thyroid cancer and positive lymph node metastasis were 20.0% (3/15), 61

  20. Survivin isoforms and clinicopathological characteristics in colorectal adenocarcinomas using real-time qPCR

    Institute of Scientific and Technical Information of China (English)

    Anastasia Pavlidou; Maria Dalamaga; Christos Kroupis; George Konstantoudakis; Maria Belimezi; George Athanasas; Kleanthi Dimas

    2011-01-01

    AIM: To investigate three isoforms of survivin in colorectal adenocarcinomas. METHODS: We used the LightCycler Technology (Roche), along with a common forward primer and reverse primers specific for the splice variants and two common hybridization probes labeled with fluorescein and LightCycler- Red fluorophore (LC-Red 640). Real time quantitative polymerase chain reaction (PCR) was performed on cDNAs from 52 tumor specimens from colorectal cancer patients and 10 unrelated normal colorectal tissues. In the patients group, carcinoembryonic antigen (CEA) and CA19-9 tumor markers were also measured immunochemically. RESULTS: Wild type survivin mRNA isoform was expressed in 48% of the 52 tumor samples, survivin-2b in 38% and survivin-ΔΕx3 in 29%, while no expression was found in normal tissues. The mRNA expression of wild type survivin presented a significant correlation with the expression of the ratio of survivin-2b, survivin-ΔΕx3, survivin-2b/wild type survivin and survivin-ΔΕx3/wild type survivin (P < 0.001). The mRNA expression of wildsurvivin and survivin-ΔΕx3 was related with tumor size and invasion (P = 0.006 and P < 0.005, respectively). A significant difference was found between survivin-2b and morphologic cancer type. Also, the ratio of survivin-ΔEx3/ wild-survivin was significantly associated with prognosis. No association was observed between the three isoforms and grade, metastasis, Dukes stage and gender. The three isoforms were not correlated with CEA and CA19-9. CONCLUSION: Survivin isoforms may play a role in cell apoptosis and their quantification could provide information about clinical management of patients suffering from colorectal cancer.

  1. Dentatin Induces Apoptosis in Prostate Cancer Cells via Bcl-2, Bcl-xL, Survivin Downregulation, Caspase-9, -3/7 Activation, and NF-κB Inhibition

    Directory of Open Access Journals (Sweden)

    Ismail Adam Arbab

    2012-01-01

    Full Text Available This study was set to investigate antiproliferative potential of dentatin (a natural coumarin isolated from Clausena excavata Burm. F against prostate cancer and to delineate the underlying mechanism of action. Treatment with dentatin dose-dependently inhibited cell growth of PC-3 and LNCaP prostate cancer cell lines, whereas it showed less cytotoxic effects on normal prostate epithelial cell line (RWPE-1. The inhibitory effect of dentatin on prostate cancer cell growth was due to induction of apoptosis as evidenced by Annexin V staining and cell shrinkage. We found that dentatin-mediated accumulation of reactive oxygen species (ROS and downregulated expression levels of antiapoptotic molecules (Bcl-2, Bcl-xL, and Survivin, leading to disruption of mitochondrial membrane potential (MMP, cell membrane permeability, and release of cytochrome c from the mitochondria into the cytosol. These effects were associated with induction of caspase-9, -3/7 activities, and subsequent DNA fragmentation. In addition, we found that dentatin inhibited TNF-α-induced nuclear translocation of p65, suggesting dentatin as a potential NF-κB inhibitor. Thus, we suggest that dentatin may have therapeutic value in prostate cancer treatment worthy of further development.

  2. Survivin在后发性白内障动物模型中的表达%Expression of survivin in animal model of posterior capsule opacification

    Institute of Scientific and Technical Information of China (English)

    刘淑君; 代秀玉; 刘少义; 王文亭; 李元彬

    2014-01-01

    Objective To establish animal model of posterior capsule opacification ( PCO) in New Zealand white rab-bits and detect survivin expression in PCO tissue.Methods Thirty adult New Zealand white rabbits were randomly divided into control group and experimental group.The 5 rabbits in control group were executed.Ultrasonic phacoemul-sification was performed in the 25 experimental rabbits'eyes.Their eyes were examined by slit lamp microscope to observe the development of PCO immediately and 3, 7, 14 and 28 days after surgery, and then they were executed.The posterior capsule of right eyes was taken in all the rabbits.Western blotting and reverse transcription polymerase ( RT-PCR) were performed to detect survivin expression in PCO at different time points postoperatively.Results Both Western blotting and RT-PCR method indicated that different level of survivin expression was detected in the tissue of PCO 3, 7, 14 and 28 days after surgery.It reached the peak at the 7th day, and began to decrease at the 14th day after surgery.Conclusion The survivin expression can be detected in the tissue of PCO in certain time, which indicated that survivin is correlated with the pathogenesis of PCO.%目的:建立新西兰大白兔后发性白内障( PCO)动物模型,检测凋亡抑制因子Survivin在PCO中的表达,探讨Survivin在PCO形成过程中的作用。方法选用健康新西兰大白兔30只,随机分为对照组(5只)、实验组(25只)。对照组直接处死;实验组行右眼晶状体超声乳化吸出术,分别于术后即刻,3、7、14、28 d(每个时间点5只)对术眼行裂隙灯显微镜检查,然后处死。获取对照组及实验组右眼撕除部分前囊膜的囊袋,采用Western blotting和逆转录聚合酶链反应( RT-PCR)方法检测Survivin在对照组及实验组不同时间点PCO中的表达。结果 Western blotting和RT-PCR在对照组及实验组术后即刻均未检测到Survivin的表达,实验组术后3

  3. Effect of survivin siRNA on biological behaviour of breast cancer MCF7 cells

    Institute of Scientific and Technical Information of China (English)

    Hao Wang; Yi-Feng Ye

    2015-01-01

    Objective:To investigate the expression of survivin in breast cancer cell lines and explore the effect of survivin siRNA on biology behavior of breast cancer cells.Methods: Western blot was performed to detect the expression of survivin in breast cancer cell lines. Eukaryotic expression vector pIRES2-EGFP-Survivin siRNA was constructed and transfected in MCF7 cells with liposome, the efficiency of survivin siRNA was measured by Western blot and RT-PCR. Cell proliferation and apoptosis were detected by CCK8 and cell flow respectively. Cell migration and invasion was measured by transwell assay.Results: Survivin was highly expressed in MCF-7. Green fluorescence was found in MCF-7 cells tranfected with survivin siRNA and control siRNA by inverted fluorescence microscopy, the protein and mRNA level of survivin was significantly lower in cells tranfected with survivin siRNA compared with control group. Compared with control group, interfering the expression of survivin by siRNA significantly decreased the proliferation, migration and invasion of MCF-7 cells, the percentage of apoptosis cells was greatly promoted.Conclusions: Interfering the expression of Survivin can inhibit the cell proliferation, migration and invasion, and promot apoptosis in MCF-7.

  4. CyclinD1 and Survivin expression in parotid gland tumors%CyclinD1和Survivin在腮腺肿瘤中的表达

    Institute of Scientific and Technical Information of China (English)

    李云杉

    2014-01-01

    ObjectiveTo explore the cell cycle protein (CyclinD1) and apoptosis inhibiting factor (Survivin) expression in parotid gland tumors in the relationship.MethodsSelect from October 19, 2012 to 2012 on July 19 days the hospital for treatment of 54 patients with parotid gland tumor pathological section. ResultsCyclinD1 in the normal group, benign tumor and malignant tumor group, the positive rate of 5.0%, 25.0% and 70.6%, respectively, expression increased obviously, the difference was statistically significant (P<0.05), Survivin in the normal group, benign tumor and malignant tumor group were 0.0%, 30.0% and 67.6%, respectively, to express obviously increased, the difference was statistically significant (P<0.05). ConclusionCyclinD1 and Survivin in parotid gland tumor development played a synergy, can be used as important reference for diagnosis and treatment of parotid gland.%目的:探究细胞周期蛋白(CyclinD1)和凋亡抑制因子(Survivin)在腮腺肿瘤中的表达关系。方法选取自2012年10月19日~2014年7月19日来我院进行治疗的54例腮腺肿瘤患者的病理切片。结果 CyclinD1在常人组、良性肿瘤组和恶性肿瘤组的阳性率分别为5.0%、25.0%和70.6%,表现明显增高,差异有统计学意义(P<0.05),Survivin在常人组、良性肿瘤组和恶性肿瘤组的阳性率分别为0.0%、30.0%和67.6%,表达明显增高,差异有统计学意义(P<0.05)。结论 CyclinD1与Survivin在腮腺肿瘤的发展中起到了协同作用,可作为腮腺诊治的重要参考依据。

  5. 胃癌变过程中Hp感染与凋亡基因Survivin表达的关系%Relationship between helicobacter pylori infection and expression of survivin gene in carcinogenesis of gastric mucosa

    Institute of Scientific and Technical Information of China (English)

    刘爱群; 葛莲英; 罗元; 林思彤

    2011-01-01

    Objective To study the relationship between helicobacter pylori(Hp) infection and the expression of survivin gene and cell apoptosis in carcinogenesis of gastric mucosa. Methods Hp infections were evaluated by rapid urease test, methylene blue and Warthin-Starry staining method, and the expression of survivin was detected by immunohistochemical staining, while the level of cell apoptosis was detected by TUNEL staining, including 62 cases of chronic superficial gastris (CSG), 55 cases chronic atrophic gastris(CAG), 52 cases intestinal melaplasia(IM), 46 cases atypical hyperplasia(AH), 65 cases gastric carcinoma(GC). Results The infection rates of Hp and the positive rates of survivin expression increased with the degree of carcinogenesis of gastric mucosa. The infection rate of Hp in CAG, IM, AH, GC were higher than that in CSC respectively( P <0. 05), and the infection rate of Hp in GC was higher than that in CAG( P < 0.05). The expressions of survivin in AH, GC were higher than those in IM and CAG respectively( P < 0.01 ). The positive rate of survivin expression in AH and GC with Hp positive were higher than those with Hp negative( P < 0.05 ). the apoptosis indexes with survivin positive expression in CAG, IM, AH and GC were lower than those with survivin negative( P < 0.05 ). The expression of survivin protein in GC with Hp infections was closely correlate with tissue differentiation, The expression level of survivin protein in undifferentiated or poorly-differentiated GC was much higer than that in well-differen-tiated ones(P < 0.05). Conclusion Hp infection can gradually up-regulate the exprssion of survivin gene during the course of gastric cancer prosession, and inhibit the cell apoptosis and differentiation to carcinogensis.%目的 研究胃癌变过程中幽门螺杆菌(Hp)感染与凋亡基因Survivin表达、细胞凋亡的关系.方法 用快速尿素酶法、W-S银染法和美蓝法联合检测62例慢性浅表性胃炎(CSG)、55

  6. Phase I clinical study of anti-apoptosis protein, survivin-derived peptide vaccine therapy for patients with advanced or recurrent colorectal cancer

    Directory of Open Access Journals (Sweden)

    Minamida Hidetoshi

    2004-06-01

    Full Text Available Abstract Survivin is a member of the inhibitor of apoptosis protein (IAP family containing a single baculovirus IAP repeat domain. It is expressed during fetal development but becomes undetectable in terminally differentiated normal adult tissues. We previously reported that survivin and its splicing variant survivin-2B was expressed abundantly in various types of tumor tissues as well as tumor cell lines and was suitable as a target antigen for active-specific anti-cancer immunization. Subsequently, we identified an HLA-A24-restricted antigenic peptide, survivin-2B80-88 (AYACNTSTL recognized by CD8+ cytotoxic T lymphocytes (CTLs. We, therefore, started a phase I clinical study assessing the efficacy of survivin-2B peptide vaccination in patients with advanced or recurrent colorectal cancer expressing survivin. Vaccinations with survivin-2B peptide were given subcutaneously six times at 14-day intervals. Of 15 patients who finished receiving the vaccination schedule, three suffered slight toxicities, including anemia (grade 2, general malaise (grade 1, and fever (grade 1. No severe adverse events were observed in any patient. In 6 patients, tumor marker levels (CEA and CA19-9 decreased transiently during the period of vaccination. Slight reduction of the tumor volume was observed in one patient, which was considered a minor responder. No changes were noted in three patients while the remaining eleven patients experienced tumor progression. Analysis of peripheral blood lymphocytes of one patient using HLA-A24/peptide tetramers revealed an increase in peptide-specific CTL frequency from 0.09% to 0.35% of CD8+ T cells after 4 vaccinations. This phase I clinical study indicates that survivin-2B peptide-based vaccination is safe and should be further considered for potential immune and clinical efficacy in HLA-A24-expression patients with colorectal cancer.

  7. 左炔诺孕酮下调survivin基因促进人子宫肌瘤细胞凋亡%Levonorgestrel induces apoptosis through down-regulation of survivin gene expression in human uterine leiomyoma cells

    Institute of Scientific and Technical Information of China (English)

    仇黎丽; 徐青; 徐昌芬

    2012-01-01

    Objective To explore the function of Survivin in levonorgestrel ( LNG) -induced apoptosis in cultured human uterine leiomyoma cells (UtLMC) in vitro. Methods AO/EB double staining was applied to discriminate the apoptotic cells from dead ones. The changes of survivin mRNA expressions in UtLMC after LNG treatment were detected by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method. Furthermore, Western blot was used to determine survivin protein expression in LNG-induced UtLMC apoptosis. Results The result of AO/EB double staining showed that there were more viable apoptotic cells in LNG-treated groups than control group and the number of non-viable apoptotic cells and dead cells increased while the concentration of LNG elevated. Survivin expression in LNG-treated cells was decreased both at mRNA and protein levels according to RT-PCR and Western blot results. Conclusions Levonorgestrel treatment may induce a substantial apoptotic response in UtLMC, which was enhanced with the raise of levonorgestrel concentration. It may be due to the marked down-regulation of survivin gene expression.%目的 探讨左炔诺孕酮(LNG)诱导人子宫肌瘤细胞(UtLMC)凋亡过程中survivin的表达变化.方法 原代培养人子宫肌瘤细胞传代后,加入不同浓度LNG,以AO/EB双染法区分早、晚期凋亡细胞和坏死细胞;RT-PCR检测抑凋亡基因survivin mRNA的表达,Western blot测定survivin蛋白的表达.结果 10 mg/L LNG作用UtLMC后早期凋亡细胞多于阴性对照组,随着LNG剂量的增加,晚期凋亡细胞也逐渐增多,核浓聚、偏位,被染成桔红色;在10 mg/L以上LNG诱导的UtLMC凋亡中,survivin mRNA表达显著下降,蛋白表达由对照组的33.82±0.02下降至10.37±0.03 (P <0.05).结论 一定浓度的左炔诺孕酮所诱导的人子宫肌瘤细胞凋亡,可能与抑制survivin抗凋亡活性相关.

  8. A cell-permeable dominant-negative survivin protein induces apoptosis and sensitizes prostate cancer cells to TNF-α therapy

    Directory of Open Access Journals (Sweden)

    Kanwar Jagat R

    2010-10-01

    Full Text Available Abstract Background Survivin is a member of the inhibitor-of-apoptosis (IAP family which is widely expressed by many different cancers. Overexpression of survivin is associated with drug resistance in cancer cells, and reduced patient survival after chemotherapy and radiotherapy. Agents that antagonize the function of survivin hold promise for treating many forms of cancer. The purpose of this study was to investigate whether a cell-permeable dominant-negative survivin protein would demonstrate bioactivity against prostate and cervical cancer cells grown in three dimensional culture. Results A dominant-negative survivin (C84A protein fused to the cell penetrating peptide poly-arginine (R9 was expressed in E. coli and purified by affinity chromatography. Western blot analysis revealed that dNSurR9-C84A penetrated into 3D-cultured HeLa and DU145 cancer cells, and a cell viability assay revealed it induced cancer cell death. It increased the activities of caspase-9 and caspase-3, and rendered DU145 cells sensitive to TNF-α via by a mechanism involving activation of caspase-8. Conclusions The results demonstrate that antagonism of survivin function triggers the apoptosis of prostate and cervical cancer cells grown in 3D culture. It renders cancer cells sensitive to the proapoptotic affects of TNF-α, suggesting that survivin blocks the extrinsic pathway of apoptosis. Combination of the biologically active dNSurR9-C84A protein or other survivin antagonists with TNF-α therapy warrants consideration as an approach to cancer therapy.

  9. Noscapine induced apoptosis via downregulation of survivin in human neuroblastoma cells having wild type or null p53.

    Directory of Open Access Journals (Sweden)

    Shiwang Li

    Full Text Available Neuroblastoma is the most common extracranial solid tumor of childhood. It accounts for 15% of pediatric cancer deaths. Chemotherapy is the mainstay of treatment in children with advanced neuroblastoma. Noscapine, a nontoxic natural compound, can trigger apoptosis in many cancer types. We now show that p53 is dispensable for Noscapine-induced cell death in neuroblastoma cell lines, proapoptotic response to this promising chemopreventive agent is mediated by suppression of survivin protein expression. The Noscapine treatment increased levels of total and Ser(15-phosphorylated p53 protein in SK-SY5Y cells, but the proapoptotic response to this agent was maintained even after knockdown of the p53 protein level. Exposure of SK-SY5Y and LA1-5S cells to Noscapine resulted in a marked decrease in protein and mRNA level of survivin as early as 12 hours after treatment. Ectopic expression of survivin conferred statistically significant protection against Noscapine-mediated cytoplasmic histone-associated apoptotic DNA fragmentation. Also, the Noscapine-induced apoptosis was modestly but statistically significantly augmented by RNA interference of survivin in both cell lines. Furthermore, Noscapine-induced apoptotic cell death was associated with activation of caspase-3 and cleavage of PARP. In conclusion, the present study provides novel insight into the molecular circuitry of Noscapine-induced apoptosis to indicate suppression of survivin expression as a critical mediator of this process.

  10. Noscapine induced apoptosis via downregulation of survivin in human neuroblastoma cells having wild type or null p53.

    Science.gov (United States)

    Li, Shiwang; He, Jing; Li, Shuai; Cao, Guoqing; Tang, Shaotao; Tong, Qiangsong; Joshi, Harish C

    2012-01-01

    Neuroblastoma is the most common extracranial solid tumor of childhood. It accounts for 15% of pediatric cancer deaths. Chemotherapy is the mainstay of treatment in children with advanced neuroblastoma. Noscapine, a nontoxic natural compound, can trigger apoptosis in many cancer types. We now show that p53 is dispensable for Noscapine-induced cell death in neuroblastoma cell lines, proapoptotic response to this promising chemopreventive agent is mediated by suppression of survivin protein expression. The Noscapine treatment increased levels of total and Ser(15)-phosphorylated p53 protein in SK-SY5Y cells, but the proapoptotic response to this agent was maintained even after knockdown of the p53 protein level. Exposure of SK-SY5Y and LA1-5S cells to Noscapine resulted in a marked decrease in protein and mRNA level of survivin as early as 12 hours after treatment. Ectopic expression of survivin conferred statistically significant protection against Noscapine-mediated cytoplasmic histone-associated apoptotic DNA fragmentation. Also, the Noscapine-induced apoptosis was modestly but statistically significantly augmented by RNA interference of survivin in both cell lines. Furthermore, Noscapine-induced apoptotic cell death was associated with activation of caspase-3 and cleavage of PARP. In conclusion, the present study provides novel insight into the molecular circuitry of Noscapine-induced apoptosis to indicate suppression of survivin expression as a critical mediator of this process.

  11. 大鼠肝部分切除后肝再生组织中Survivin基因的表达变化%Expression of Survivin gene in hepatic regeneration tissue of rat after partial hepatectomy

    Institute of Scientific and Technical Information of China (English)

    金玉姬; 王德欣; 陈晓黎; 仉海峰; 叶素素; 侯青顺

    2012-01-01

    目的:探讨Survivin在大鼠肝部分切除(Partial hepatectomy,PH后肝再生过程中的作用及其意义.方法:用70%肝部分切除方法,建立Wister大鼠肝再生模型,在肝再生不同时间点,采用免疫组化及Western blot方法,检测肝组织中Survivin和细胞周期相关蛋白CyclinD1的表达.结果:在肝切除术后12~72 h,Survivin蛋白表达量明显增高,且在48 h达到峰值,细胞周期相关蛋白CyclinD1在术后12、24、48、72 h均表达上调,与Survivin蛋白表达具有高度相关性.结论:Survivin蛋白的高表达与肝部分切除后肝再生过程密切相关.%Objective:To investigate the effects and its significance of survivin gene in rat liver regeneration after partial hepatectomy. Methods: Wister rat liver regeneration model of 70% partial hepatectomy was established. The expressions of Survivin and cell cycle-related protein cyclinD1 were detected by immunohistochemistry in liver regeneration rat tissues at different time points. Results: The level of Survivin protein expression was significantly increased during 12-72 h after partial hepatectomy and reached its peak at 48 h after partial hepatectomy. The level of cyclinD1 was upregulated at 12,24,48 h,and 72 h after partial hepatectomy and highly correlated with survivin protein level. Conclusion:The high expression of Survivin protein is significantly correlated with the liver regeneration after partial hepatectomy.

  12. Research of the relationship between Survivin and Caspase-9 expression in colorectal cancer%Survivin、Caspase-9的表达与大肠癌关系的研究进展

    Institute of Scientific and Technical Information of China (English)

    赵岩; 王志强

    2015-01-01

    大肠癌是当今严重威胁人类健康的疾病之一,它是多因素协同作用的结果,其中调控凋亡的基因异常表达在大肠癌发生、发展中起重要促进作用。Survivin和Caspase-9在调控细胞凋亡过程中存在关联,它们的异常激活及失活都与大肠癌的发生密切相关。先前的研究表明Survivin和Caspase-9与大肠癌的浸润、分化、分期、淋巴结转移的关系依然存在争议。两者在大肠癌的诊断、预后及靶向治疗方面具有应用前景。%Colorectal cancer is one of the most serious threaten disease to human health. It is the result of many factors, the abnormal expression of the genes which regulate apoptosis play an important role in the development of colorectal cancer. Survivin and Caspase-9 are associated with the regulation of apoptosis, their abnormal activation and inactivation are closely related to the occurrence of colorectal cancer. Previous studies have indicated that the relationship between Caspase-9、Survivin and the invasion, differentiation, staging and lymph node metastasis of colorectal cancer remain exist controversy. There are clnical value in both of them in the aspects of diagnosis, prognosis and targeted therapy colorectal cancer.

  13. Arctigenin promotes apoptosis in ovarian cancer cells via the iNOS/NO/STAT3/survivin signalling.

    Science.gov (United States)

    Huang, Ke; Li, Li-an; Meng, Yuan-guang; You, Yan-qin; Fu, Xiao-yu; Song, Lei

    2014-12-01

    Arctigenin is a biologically active lignan extracted from the seeds of Arctium lappa and shows anticancer activity against a variety of human cancers. The aim of this study was to determine the effects of arctigenin on ovarian cancer cell proliferation and survival and associated molecular mechanisms. Human ovarian cancer OVCAR3 and SKOV3 cells were treated with arctigenin, and cell proliferation and apoptosis were assessed. Western blot analysis was used to examine signal transducer and activator of transcription-3 (STAT3) phosphorylation and survivin and inducible nitric oxide synthase (iNOS) expression. The involvement of STAT3/survivin/iNOS/NO signalling in arctigenin action was checked. Arctigenin treatment resulted in a significant and dose-dependent inhibition of cell proliferation. Arctigenin-treated cells showed a 4-6 times increase in the percentage of apoptosis, compared with control cells. Pre-treatment with Ac-DEVD-CHO, a specific inhibitor of caspase-3, counteracted the induction of apoptosis by arctigenin. Arctigenin treatment significantly inhibited STAT3 phosphorylation and survivin and iNOS expression. Arctigenin-induced apoptosis was impaired by pre-transfection with survivin-expressing plasmid or addition of chemical nitric oxide (NO) donors. Additionally, exogenous NO prevented the suppression of STAT3 phosphorylation and survivin expression by arctigenin. Arctigenin treatment inhibits the proliferation and induces caspase-3-dependent apoptosis of ovarian cancer cells. Suppression of iNOS/NO/STAT3/survivin signalling is causally linked to the anticancer activity of arctigenin. Therefore, arctigenin may be applicable to anticancer therapy for ovarian cancer.

  14. 联合检验尿Livin、Survivin mRNA表达在膀胱癌早期诊断中的价值%Value of combined detection of urinary Livin and Survivin mRNA expression in the early diagnosis of bladder cancer

    Institute of Scientific and Technical Information of China (English)

    江龙来; 谢梅茂; 王晓荣; 赵雁; 王忠军

    2013-01-01

    Objective To evaluate the value of urine Livin and Survivin mRNA expression in the early diagnosis of bladder cancer.Methods Fifty-two cases of early bladder cancer and 30 cases of nonurinary system tumors were selected for the combined detection of urinary Livin and Survivin and urine cytology.Results Livin and Survivin in urine and urinary cytology sensitivity were 71.2% (37/52),67.3% (35/52),23.1% (12/52) ; Specificity were 96.7% (29/30),93.3% (28/30) and 96.7% (29/30) respectively.Urine Survivin sensitivity,specificity compared with Livin were no significant difference (all P > 0.05).Urinary Livin and Survivin were more sensitive than urine cytology,and the differences were statistically significant(all P <0.05).Conclusion The combined urinary detection of Livin,Survivin mRNA expression is a noninvasive early diagnosis of bladder cancer with sensitivity and specificity.%目的 评价尿凋亡抑制因子Livin、Survivin mRNA表达在膀胱癌早期诊断中的应用价值.方法 选择南昌大学第四附属医院2008至2012年收治的52例早期膀胱尿路上皮癌和30例非泌尿系统肿瘤患者,采用PCR的方法对研究对象的尿液进行Livin和Survivin检测,并行尿脱落细胞学检查.结果 病例组和对照组尿Livin和Survivin及尿脱落细胞学检查阳性分别为37、35、12例,1、2、1例;敏感度分别为71.2%、67.3%、23.1%;特异度分别为96.7%,93.3%和96.7%;尿Survivin的敏感度、特异度和Livin比较差异均无统计学意义(均P>0.05);尿Livin和Survivin的敏感度均高于尿脱落细胞学,差异均有统计学意义(均P<0.05).结论 联合检测尿Livin、Survivin mRNA表达是无创性早期诊断膀胱癌敏感和特异的指标.

  15. Survivin: Potential role in diagnosis, prognosis and targeted therapy of gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Ting-Ting Wang; Xiao-Ping Qian; Bao-Rui Liu

    2007-01-01

    Survivin is a protein that is highly expressed in a vast number of malignancies, but is minimally expressed in normal tissues. It plays a role as an inhibitor of cell death in cancer cells, thus facilitating the growth of these cells.Tn the case of gastric cancer, survivin is over-expressed in tumor cells and plays a role in the carcinogenesis process. Several studies on gastric cancer have indicated that there is a relationship between survivin expression and the ultimate behavior of the carcinoma. Since the expression pattern of survivin is selective to cancer cells,it has been described as an "ideal target" for cancer therapy. Currently, several pre-clinical and clinical trials are on-going to investigate the effects of interfering with survivin function in cancer cells as a biologic therapy. Survivin is a potentially significant protein in the diagnosis, prognosis and treatment of gastric tumors.

  16. Survivin和MMP-2在宫颈鳞癌患者中的表达及其临床意义%Expressions and clinical significances of Survivin and MMP-2 in patients with cervical squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    王爱平

    2011-01-01

    目的:探讨Survivin和MMP-2在宫颈鳞癌中的表达及其临床意义.方法:对45例宫颈CINⅠ-Ⅱ、18例宫颈GINⅢ和33例宫颈浸润性鳞癌患者分别进行Survivin和MMP-2检查,比较各组患者的阳性表达,并分析其与宫颈病变级别的相关性.结果:Survivin和MMP-2的阳性表达与宫颈病变的级别呈正相关,不同宫颈病变级别的阳性率比较差异均有统计学意义(P<0.05,P<0.01).结论:临床宫颈病变患者活检后应行Survivin和MMP-2检查,可提示其预后并指导临床治疗.%Objective: To explore the expressions and clinical significances of Survivin and MMP - 2 in patients with cervical squa-mous cell carcinoma Methods: The expression levels of Survivin and MMP -2 in 45 cases with cervical intraepithelial neoplasia (CIN) I ~ II , 18 cases with CINM and 33 cases with invasive cervical squamous cell carcinoma were detected, the positive expression levels of Survivin and MMP - 2 in different groups were compared, its correlation with the degree of cervical lesions was analyzed. Results: There was a positive correlation between the positive expression levels of Survivin and MMP - 2 and the degree of cervical lesions, there was significant difference in the positive expression levels of Survivin and MMP - 2 among cervical lesions of different degrees ( P < 0. 05, P <0. 01) . Conclusion; For the patients with cervical lesions, Survivin and MMP -2 detection should be carried out after biopsy, which can indicate prognosis and direct clinical treatment

  17. Survivin与PTEN在喉部鳞状细胞癌变中的表达关系初探%Analysis on expression of Survivin and PTEN in vocal cords polyps,papilloma of larynx and laryngeal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    郭强中; 李云英

    2012-01-01

    Through exploring the differentiation on positive expressing rates between oncogene Survivin and tumor-suppresser gene PTEN(phosphatase and tensin homolog deleteted on chromosome Ten) on vocal cord polyps(VCP), adult type laryngeal papilloma (ALP) , and Laryngeal Squamous Cell Carcinoma (LSCC) , to reveal the mechanism in canceration of human laryngeal squamous cells, from benign proliferative lesion, benign tumor to malignant tumor in larynx. Method:After picking out 18 cases of VCP, 10 cases of ALP, and 18 cases of LSCC for immunohistochemical process of Survivin and PTEN with two continuous section preparations, the differentiations of positive expression rates of Survivin and PTEN in the same human laryngeal squamous cells areas among three diseases were compared. Result:The positive expressing rates of Survivin and PTEN in VCP were obviously more lower than in ALP and LSCC(P0.05). The positive expressing rates of Survivin were always higher than PTEN in VCP, ALP and LSCC evi-dently(P<0. 05). Conclusion:PTEN, expressing for competition purpose, shows a subordinative relationship with Survivin. Although they have opposite functions in determine whether the canceration of laryngeal squamous cells take place or not, Survivin is always playing the leading role and making predominant impact on development of benign proliferative lesion, benign and malignant tumor in larynx.%目的:研究声带息肉、成人型喉乳头状瘤和喉鳞状细胞癌3种疾病中Survivin和PTEN的表达特点,以了解喉鳞状细胞从增生到癌变的可能机制.方法:选取符合临床与病理诊断的声带息肉患者18例、成人型喉乳头状瘤患者10例和喉鳞状细胞癌患者18例,将每个患者的病理蜡块标本各连续切片2张,分别进行Survivin和PTEN免疫组织化学检测.比较致癌基因Survivin和抑癌基因PTEN在这3种疾病同一区域鳞状上皮细胞中的表达差异.结果:声带息肉的Survivin和PTEN阳性率均显著低于喉乳头

  18. Matrine induces mitochondrial apoptosis in cisplatin-resistant non-small cell lung cancer cells via suppression of β-catenin/survivin signaling.

    Science.gov (United States)

    Wang, Huan-Qin; Jin, Jian-Jun; Wang, Jing

    2015-05-01

    Matrine is an alkaloid isolated from Sophora flavescens and shows anticancer activities. The present study was carried out to determine the cytotoxic effects of matrine on cisplatin-resistant non-small cell lung cancer (NSCLC) cells and the associated molecular mechanisms. Parental and cisplatin-resistant A549 and H460 NSCLC cells were treated with 1 or 2 g/l of matrine for 48 h, and cell viability and apoptosis were assessed. β-catenin-mediated transcriptional activity, mitochondrial membrane potential (ΔΨm) changes, activation of caspases, and survivin expression were examined. The effect of overexpression of survivin on the anticancer activity of matrine was investigated. Compared to the parental cells, cisplatin-resistant NSCLC cells showed increased β-catenin transcriptional activity. Matrine treatment resulted in a significant reduction in β-catenin activation and survivin expression in the cisplatin-resistant cells. Matrine caused apoptotic death in the cisplatin-resistant NSCLC cells, coupled with loss of ΔΨm and activation of caspase-9 and -3. Matrine-induced apoptosis of the cisplatin-resistant NSCLC cells was significantly reversed by overexpression of survivin. In conclusion, matrine exposure induces mitochondrial apoptosis in cisplatin-resistant NSCLC cells, which is largely mediated through inactivation of β-catenin/survivin signaling. Further investigation of the therapeutic benefit of matrine in overcoming cisplatin resistance in NSCLC is warranted.

  19. Selection of optimal antisense accessible sites of survivin and its application in treatment of gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Qiang-Song Tong; Li-Duan Zheng; Fang-Min Chen; Fu-Qing Zeng; Liang Wang; Ji-Hua Dong; Gong-Cheng Lu

    2005-01-01

    AIM: To select the optimal antisense accessible sites of survivin, a highly expressed gene in tumor tissues, in order to explore a novel approach to improve biological therapy of gastric cancer.METHODS: The 20 mer random oligonucleotide library was synthesized, hybridized with in vitro transcribed total survivin cRNA, then digested by RNase H. After primer extension and autoradiography, the antisense accessible sites (AAS) of survivin were selected. Then RNADraw software was used to analyze and choose the AAS with obvious stem-loop structures, according to which the complementary antisense oligonucleotides (AS-ODNs) were synthesized and transferred into survivin highly- expressing gastric cancer cell line MKN-45. Survivin expression was detected by RT-PCR and Western Blotting. Cellular growth activities were assayed by tetrazolium bromide (MTT)colorimetry. Cellular ultrastructure was observed by electronic microscopy, while apoptosis was detected by annexin V-FITC and propidium iodide staining flow cytometry.RESULTS: Thirteen AAS of survivin were selected in vitro.Four AAS with stem-loop structures were chosen, locating at 207-226 bp, 187-206 bp, 126-145 bp and 44-63 bp of survivin cDNA respectively. When compared with nontranfection controls, their corresponding AS-ODNs (AS-ODN1,AS-ODN2, AS-ODN3 and AS-ODN4) could reduce Survivin mRNA levels in MKN-45 cells by 54.3±1.1% (t= 6.12, P<0.01),86.1±1.0% (t= 5.27, P<0.01), 32.2±1.3% (t= 7.34, P<0.01)and 56.2±0.9% (t= 6.45, P<0.01) respectively, while survivin protein levels were decreased by 42.2±2.5% (t = 6.26,P<0.01), 75.4±3.1% (t= 7.11, P<0.01), 28.3±2.0% (t= 6.04,P<0.01) and 45.8±1.2% (t = 6.38, P<0.01) respectively.After transfection with 600 nmol/L AS-ODN1~AS-ODN4 for24 h, cell growth was inhibited by 28.12±1.54% (t= 7.62,P<0.01), 38.42±3.12% (t = 7.75, P<0.01), 21.46±2.63%(t= 5.94, P<0.01) and 32.12±1.77% (t= 6.17, P<0.01)respectively. Partial cancer cells presented the

  20. Expression and comparative research of Survivin between Uygur and Han women in different cervical diseases%中国维汉族妇女宫颈病变中Survivin的表达及对比研究

    Institute of Scientific and Technical Information of China (English)

    魏薇; 周瑾

    2012-01-01

    Objective:To explore the relationship between expression of apoplosis-inhibited protein Survivin and the pathogensis and progression of cervical cancer, to compare the difference between Uygur and Han women. Methods;To collect 30 cases of chronic cervicitis of each nation,30 cases of low-grade squamous intraepithelial lesion in each nation ,30 cases of high-grade cervical squamous intraepithelial neoplasia each nation and 30 cases of invasive cervical cancer each nation and to detect the expression of survivin with immunohistochem technique. Results: The positive position of Survivin protein was inside cell cytoplasm and nuclei. The expression rates of Survivin were increased gradually with cervical disease pathogenesis. The positive rate of Survivin was not correlated with high-grade cervical squamous intraepithelial neoplasia and invasive cervical cancer ( P > 0. 05), and the differences were not correlated between Uygur and Han women ( P > 0.05 ). There was no difference of Surviving expression between Uygur and Han women. Conclusion:The abnormal expression of Survivin protein waa related closely with cervical cancer and pre-cancerous, Survivin appears in the early time of malignant transformation, and it promote the occurrence and development of cervical cancer. Survivin may become a target for prognosis and as a gene for targeting treatment.%目的:探究凋亡抑制蛋白Survivin与宫颈癌发生、发展的关系,对比Survivin的表达在维、汉族之间是否存在族群间差异.方法:收集维吾尔族和汉族妇女宫颈炎(各30例)、宫颈低级别上皮内瘤变(CIN Ⅰ各30例)、高级别上皮内瘤变(CINⅡ-Ⅲ各30例)及宫颈浸润性鳞癌(各30例)石蜡包埋组织标本共240例,采用免疫组织化学SP法.结果:Survivin蛋白的阳性表达定位于细胞质和(或)胞核内,表达强度随病变程度加重呈递增趋势,但CINⅡ-Ⅲ组与宫颈癌组的阳性率无差异(P>0.05),维汉组间比

  1. Notch-1和Survivin在甲状腺乳头状癌中的表达及意义%Expression and clinical significance of Notch-1 and Survivin in papillary thyroid carcinoma

    Institute of Scientific and Technical Information of China (English)

    史永亮

    2013-01-01

    目的 探讨Notch-1、Survivin在甲状腺乳头状癌组织中的表达及临床意义.方法 收集80例甲状腺乳头状癌、70例甲状腺腺瘤和50例正常甲状腺组织石蜡标本,采用免疫组织化学SP法检测Notch-1、Survivin蛋白的表达水平.结果 在甲状腺乳头状癌、甲状腺腺瘤和正常甲状腺组织中,Notch-1的阳性表达率分别为22.5%、78.6%、88.0%,Survivin的阳性表达率分别为70.0%、12.9%、0%,与甲状腺腺瘤和正常甲状腺组织比较,甲状腺乳头状癌组织Notch-1、Survivin分别呈低表达和高表达状态(P<0.05),且二者表达呈明显负相关(r=-0.512,P<0.01).Notch-1和Survivin在甲状腺乳头状癌组织的异常表达与肿瘤临床分期、淋巴结转移有关(P<0.05).结论 在甲状腺乳头状癌的侵袭转移过程中存在Notch-1、Survivin的异常表达,对二者的联合检测有助于判断疾病的预后和转归.%Objective To explore the expression and clinical significance of Notch 4 and Survivin in papillary thyroid carcinoma tissues. Methods The expression levels of Notch-1 and Survivin were detected by immunohistochemistry in 80 cases of papillary thyroid carcinoma , 70 cases of thyroid adenoma and 50 cases of normal thyroid gland tissues. The correlation between the expression levels and clinical pathological parameters was analyzed. Results The positive expression rate of Notch-1 in papillary thyroid carcinoma, thyroid adenoma and normal thyroid gland tissues was 22. 5% ,78. 6% ,88. 0% ,respectively,and the positive expression rate of Survivin was 70.0% ,12.9% ,0% ,respectively, in papillary thyroid carcinoma , thyroid adenoma and normal thyroid gland tissues. As compared with that in thyroid adenoma and normal thyroid gland tissues , Notch4 in papillary thyroid carcinoma was low-expressed, however, Survivin was over-expressed ( P < 0. 05 ) , furthermore the expression of Notch4 was closely correlated to that of Survivin ( P <0. 01). The abnormal expression of

  2. 三阴乳腺癌组织中p27、survivin的表达及临床意义%Expression of p27, survivin in triple negative breast carcinoma and its clinic significance

    Institute of Scientific and Technical Information of China (English)

    王琼; 贺玉娟; 刘玮; 付军; 刘蓓

    2012-01-01

    Objective To investigate the roles of p27 and survivin in the occurrence and development of triple negative breast carcinoma(TNBC) . Methods The tissue specimen from 56 cases of TNBC( tumor group) , infiltrating ductal carcinoma of breast (infiltrating group) , benign mammary gland hyperplasia (hyperplasia group) were collected. The protein expression of p27 and survivin was detected by immunohistochemistry, the correlation between the expression of p27, survivin and the clinicopathologic parameters of TNBC were analyzed. Results The positive expression rate of p27 in tumor group, infiltrating group and hyperplasia group was 41.1% , 67.8 % and 91.1% respectively, the rate in the tumor group was significantly lower than that in the other two groups, and which in the infiltrating group was significantly lower than that in the hyperplasia group( all P <0. 01) ; the positive expression rate of survivin was 69. 6% ,60. 7% and 5. 3% respectively,which in the tumor group and infiltrating group was significantly higher than that in the hyperplasia group(both P <0.01. The expression of p27 in TNBC was related to the patients's age, size of tumor, clinical stages and lymph node metastasis. The expression of survivin in TNBC was related to the patients'age, size of tumor. The expression of p27 was negatively correlated with the expression of survivin in TNBC. Conclusions The abnormal expression of p27 and survivin may correspond to the occurrence, development and prognosis of TNBC.%目的 探讨p27、survivin在三阴乳腺癌(TNBC)发生、发展中的作用.方法 选择我院手术切除的TNBC(肿瘤组)、非特殊类型乳腺浸润性导管癌(浸润组)、良性乳腺增生症(增生组)组织标本各56份,采用免疫组化SP二部法检测各组p27、survivin的表达情况,并分析p27、survivin的表达与TNBC临床病理参数的相关性.结果 肿瘤组、浸润组、增生组p27阳性表达率分别为41.1%、67.8%、91.1%,肿瘤组明显低于其余

  3. Interfering RNA vector of survivin:Its construction and survivin expression in HEK293 cell%Survivin干扰RNA载体的构建及其在HEK293细胞中的表达

    Institute of Scientific and Technical Information of China (English)

    倪金良; 陈晓星; 张国新; 施瑞华; 郝波

    2009-01-01

    目的:探索合成survivin siRNA和RNAi载体的构建,及其在HEK293细胞中的表达.方法:分别采用化学合成的针对survivin的siRNA和shRNA载体(pGCsi载体),转染HEK293细胞后观察HEK293细胞中survivin的RNA及蛋白表达.结果:不同序列的siRNA和shRNA转染人HEK293细胞后,survivin的RNA及蛋白表达出现不同程度的下降.结论:化学合成与shRNA载体均可有效抑制HEK293细胞的survivin基因表达,为利用survivin作为靶点治疗胰腺癌等恶性肿瘤提供了技术基础.

  4. SU-E-T-320: The Effect of Survivin Perturbation On the Radiation Response of Breast Cancer Cell Lines

    Energy Technology Data Exchange (ETDEWEB)

    Smith, D; Debeb, B; Woodward, W [Department of Radiation Oncology, MD Anderson Cancer Center, Houston, TX (United States)

    2014-06-01

    Purpose: Survivin is the smallest member of the inhibitor of apoptosis protein family and is well-known for its universal over-expression in human cancers. Due to its role in apoptosis and cellular proliferation, survivin is implicated in the radiation response in several cancer types, and antisurvivin treatments have had success as a radiation sensitizer in many preclinical cancer models. As no studies to date have reported survivin as a factor affecting radiation resistance in breast cancer models, we sought to evaluate the synergistic relationship between survivin function and irradiation in breast cancer cell lines. Methods: Information regarding survivin protein expression in breast cancer was retrieved from three public databases: Oncomine, Kaplan-Meier Plotter, and GOBO. For the in vitro studies, survivin function was compromised by transducing a non-functional mutant form (survivin-DN) into two breast cancer cell lines, the estrogen receptor-positive MCF7 and the triple-negative, inflammatory SUM149. Cell growth was compared in the survivin-DN and control populations with colony-formation assays. To assess how survivin affects radiation response, clonogenic assays were performed by irradiating the cell lines up to 6 Gy. Results: From the public databases, survivin is more highly expressed in triple-negative breast cancer compared to all other subtypes, and is prognostic of poor survival in all breast cancer patients. In MCF7, the survivin-DN population had decreased colony-formation potential; the opposite was true in SUM149. In the clonogenic assays, abrogation of survivin function radio-protected MCF7 cells in monolayer and 3D growth conditions, while SUM149 survivin-DN cells were radiosensitized in monolayer conditions. Conclusion: We observed synergy between survivin function and radiation, although the results between the two cell lines were disparate. Further investigation is required to identify the mechanism of this discrepancy, including evaluation

  5. Calprotectin induces cell death in human prostate cancer cell (LNCaP) through survivin protein alteration.

    Science.gov (United States)

    Sattari, Mina; Pazhang, Yaghub; Imani, Mehdi

    2014-11-01

    Calprotectin (CP), an abundant heterodimeric cytosolic protein of neutrophils, conveys a variety of functions such as tumor cell growth arrest and antimicrobial activity. We investigated CP activity and its possible apoptosis-inducing mechanism of action against an antiandrogen therapy-resistance prostate cancer cell line LNCaP. Cell viability and Annexin V FITC assays were performed in order to investigate its cell death activity and apoptosis, respectively. In order to address cell death inducing mechanism(s), immunocytochemistry and immunobloting analysis, reactive oxygen species (ROS) and nitric oxide (NO) measurements were performed. The effective concentration of CP against LNCaP promoting LNCaP cell death was 200 µg/mL. ROS and NO levels of cells remarkably were enhanced following treatment with 50 and 100 µg/mL of CP, respectively. Protein expression of anti-apoptotic protein survivin was significantly decreased after administration of tumor cells with CP. Our data indicate that CP regulates the LNCaP cells viability via survivin-mediated pathway and ROS and NO enhancement. Thus, inhibition of survivin expression, enhancement of ROS and NO level by CP or other similar pharmaceutical agents might be effective in lowering the malignant proliferation of human prostate cancer cells.

  6. FOXM1 targets XIAP and Survivin to modulate breast cancer survival and chemoresistance.

    Science.gov (United States)

    Nestal de Moraes, Gabriela; Delbue, Deborah; Silva, Karina L; Robaina, Marcela Cristina; Khongkow, Pasarat; Gomes, Ana R; Zona, Stefania; Crocamo, Susanne; Mencalha, André Luiz; Magalhães, Lídia M; Lam, Eric W-F; Maia, Raquel C

    2015-12-01

    Drug resistance is a major hurdle for successful treatment of breast cancer, the leading cause of deaths in women throughout the world. The FOXM1 transcription factor is a potent oncogene that transcriptionally regulates a wide range of target genes involved in DNA repair, metastasis, cell invasion, and migration. However, little is known about the role of FOXM1 in cell survival and the gene targets involved. Here, we show that FOXM1-overexpressing breast cancer cells display an apoptosis-resistant phenotype, which associates with the upregulation of expression of XIAP and Survivin antiapoptotic genes. Conversely, FOXM1 knockdown results in XIAP and Survivin downregulation as well as decreased binding of FOXM1 to the promoter regions of XIAP and Survivin. Consistently, FOXM1, XIAP, and Survivin expression levels were higher in taxane and anthracycline-resistant cell lines when compared to their sensitive counterparts and could not be downregulated in response to drug treatment. In agreement with our in vitro findings, we found that FOXM1 expression is significantly associated with Survivin and XIAP expression in samples from patients with IIIa stage breast invasive ductal carcinoma. Importantly, patients co-expressing FOXM1, Survivin, and nuclear XIAP had significantly worst overall survival, further confirming the physiological relevance of the regulation of Survivin and XIAP by FOXM1. Together, these findings suggest that the overexpression of FOXM1, XIAP, and Survivin contributes to the development of drug-resistance and is associated with poor clinical outcome in breast cancer patients.

  7. Expressions and significances of APC and Survivin protein in colorectal multiple adenomas with a family history of colorectal cancer%有大肠癌家族史的大肠多发性腺瘤患者瘤组织中APC、Survivin 蛋白的表达及意义

    Institute of Scientific and Technical Information of China (English)

    赵婧; 荆洋; 吕宗舜; 周璐

    2015-01-01

    Objective To investigate the expressions and significances of tumor suppressor gene APC and apoptosis inhibiting gene Survivin in colorectal multiple adenomas with a family history of colorectal cancer .Methods The expres-sions of APC and Survivin gene were detected by immunohistochemical staining among the cases from 20 colorectal adeno-carcinomas , 32 colorectal multiple adenomas with a family history of colorectal cancer , 32 colorectal multiple adenomas without a family history of colorectal cancer , 20 normal colorectal mucosas .Results There had differences in positive rate for expressions of APC and Survivin among the colorectal adenocarcinomas , the colorectal multiple adenomas with a family history of colorectal cancer , the colorectal multiple adenomas without a family history of colorectal cancer , the normal color-ectal mucosas, the positive rate of APC protein was 20.0%, 56.3%, 65.6%, 95.0%, respectively (P<0.01).The positive rate of Survivin protein was 75.0%, 40.6%, 28.1%, 0%, respectively (P<0.01).In colorectal multiple ade-nomas with a family history of colorectal cancer , the positive intensity of APC protein was lower and the positive intensity of Survivin protein was higher than that in the colorectal multiple adenomas without a family history of colorectal cancer ( all P<0.05).Conclusion Compared with the normal colorectal mucosas , the APC protein down-regulated expression and Survivin protein up-regulated expression in colorectal multiple adenomas .In colorectal multiple adenomas with a family his-tory of colorectal cancer , the expression of APC protein was lower and the expression of Survivin protein was higher than that in the colorectal multiple adenomas without a family history of colorectal cancer .%目的:观察有大肠癌家族史的大肠多发性腺瘤组织中中抑癌基因APC和凋亡抑制基因Survivin的表达变化,并探讨其意义。方法采用免疫组织化学方法检测20例大肠腺癌、32例有

  8. survivin和CD44v6在非小细胞肺癌中的表达及意义%Expression and significance of survivin and CD44v6 protein in non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    魏霞; 石志红; 嵇喜祥

    2009-01-01

    Objective To investigate the expression and significance of survivin and CD44v6 in non-small cell lung cancer (NSCLC) and their correlation. Methods SP immunohistochemical technique was used to detect the expression of survivin and CD44V6 protein in 53 cases of NSCLC and 13 cases of para-eaneer nor-mal tissues. Results The positive rate of survivin and CD44v6 in NSCLC was 60. 38% and 69. 81% respec-tively,which was higher than that of normal pulmonary tissues adjacent to carcinoma(P 0.05). The expression of survivin was related to TNM stages and cell differentiation (P 0. 05). The expression of CD44v6 in squamous carcinoma was significantly higher than that of adenocareinoma (P 0. 05). There was no correlation between the expression of survivin and C1)44v6 (r = -0. 058, P >0. 05). Conclusion Survivin might be used to evaluate NSCLC development;CD44v6 might be used for the differential diagnosis of squamous carcinoma in NSCLC;both of them might be helpful to predict the metastasis of NSCLC. They might be two independent events in the process of NSCLC genesis and develop-ment.%目的 探讨非小细胞肺癌(NSCLC)中survivin及CD44v6的表达及意义,以及二者的相关性.方法 采用SP免疫组化方法检测survivin及CD44v6在53例NSCLC组织、13例癌旁正常肺组织中的表达.结果 53例NSCLC癌组织中survivin、CD44v6阳性表达率分别是60.38%和69.81%,高于癌旁正常肺组织的表达(P0.05).survivin的表达与临床TNM分期及肿瘤分化程度相关(P0.05).CD44v6在鳞癌的表达率远高于腺癌(P0.05).survivin与CD44v6之间无相关性(r=-0.058,P>0.05).结论 survivin有望作为评估NSCLC病变进展的指标,CD44v6可用来鉴别诊断NSCLC中的鳞癌,二者有可能成为预测NSCLC转移的指标.survivin、CD44v6可能是NSCLC发生发展过程中的两个独立事件.

  9. A Critical Review on “Study of the Expression of Survivin & Its Splice Variants; ΔEx3, 2b and 3b as Diagnostic Molecular Markers in Breast Cancer”

    Directory of Open Access Journals (Sweden)

    HR Rahimi

    2013-01-01

    Full Text Available Although the topic of the study seems to be a novel subject and its design looks excellent, there are some points which seem to be missed or neglected by the respected authors of the paper entitled: “Study of the Expression of Survivin & Its Splice Variants; ΔEx3, 2b and 3b as Diagnostic Molecular Markers in Breast Cancer”. Through this review, it was attempted to review and criticize some of these issues which may occur in the similar corresponding researches in the future.

  10. Epstein-Barr Virus nuclear antigen 1 (EBNA1) confers resistance to apoptosis in EBV-positive B-lymphoma cells through up-regulation of survivin.

    Science.gov (United States)

    Lu, Jie; Murakami, Masanao; Verma, Subhash C; Cai, Qiliang; Haldar, Sabyasachi; Kaul, Rajeev; Wasik, Mariusz A; Middeldorp, Jaap; Robertson, Erle S

    2011-02-05

    Resistance to apoptosis is an important component of the overall mechanism which drives the tumorigenic process. EBV is a ubiquitous human gamma-herpesvirus which preferentially establishes latent infection in viral infected B-lymphocytes. EBNA1 is typically expressed in most forms of EBV-positive malignancies and is important for replication of the latent episome in concert with replication of the host cells. Here, we investigate the effects of EBNA1 on survivin up-regulation in EBV-infected human B-lymphoma cells. We present evidence which demonstrates that EBNA1 forms a complex with Sp1 or Sp1-like proteins bound to their cis-element at the survivin promoter. This enhances the activity of the complex and up-regulates survivin. Knockdown of survivin and EBNA1 showed enhanced apoptosis in infected cells and thus supports a role for EBNA1 in suppressing apoptosis in EBV-infected cells. Here, we suggest that EBV encoded EBNA1 can contribute to the oncogenic process by up-regulating the apoptosis suppressor protein, survivin in EBV-associated B-lymphoma cells.

  11. Effects of shRNA Targeting Survivin on Apoptosis of Human Retinoblastoma Cell Line Hxo-rb44 in vitro

    Institute of Scientific and Technical Information of China (English)

    WANG Guojun; HU Yanhua; LI Pengcheng

    2006-01-01

    In order to construct a recombinant plasmid containing short hairpin RNA (shRNA) targeting survivin and to investigate its effect on survivin expression and cell apoptosis of human retinoblastoma cell line Hxo-rb44 in vitro, RNA interference plasmid pSIRENS that can express shRNA of survivin was designed, constructed, and transfected into human retinoblastoma cell line Hxo-rb44.Survivin and c-Myc expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Apoptosis of Hxo-rb44 cells was assayed by Honchest33258 staining and cell growth curve was drawn. The results showed that the oligonucleotide targeting survivin was identified in pSIRENS plasmid. After pSIRENS plasmid transfected, survivin and c-Myc expression in Hxo-rb44 cells was decreased significantly. Apoptotic rate of cells was up-regulated from (3.5±1.29) % to (36.1±19.66) %. The proliferation ability of Hxo-rb44 cells was inhibited. No significant effects on survivin expression and apoptosis of the cells were found when negative control plasmid was transfected. In conclusion, the plasmid containing shRNA targeting survivin was constructed successfully. It could inhibit efficiently the expression of survivin and c-Myc in human retinoblastoma cell Hxo-rb44 in vitro. The inhibition of the expression of c-Myc might be involved in the apoptosis of Hxo-rb44 cells.

  12. EXPRESSION AND CLINICAL SIGNIFICANCE OF SURVIVIN,CASPASE-3, GST AND PGP IN NSCLC%Survivin Caspase-3 GST Pgp在非小细胞肺癌(NSCLC)中的表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    霍小东; 孙勤暖; 李国华

    2006-01-01

    目的:探讨非小细胞肺癌(non-small-cell lung cancer,NSCLC)中存活素(survivin)、谷胱甘肽S-转移酶-π(glutathions-transferase-π,GST-π)、P-糖蛋白(P-glycoprotein,Pgp)与半胱氨酸蛋白酶(Caspase-3)之间的关系,以及它们的表达与临床各病理指标之间的关系.方法:采用免疫组织化学法检测117例NSCLC中Survivin 、GST-π、Pgp的表达水平,流式细胞术检测Caspase-3的表达FI值;根据肿瘤耐药机制的不同,选择一个凋亡抑制元素、一个凋亡促进元素与两个耐药元素对肿瘤的耐药进行多方面的分析评估.结果:Survivin、GST-π、Pgp的阳性率明显高于对照组(P<0.01),Survivin、Pgp与肿瘤的分型无关(P>0.05),二者在分期分化表达中随其恶性程度增加,表达增强,而具有统计学意义(P<0.05);Pgp在淋巴结转移中高表达,与肿瘤转移有关;Survivin在淋巴结转移中的表达无显著性差异,与肿瘤转移无关;GST-π与肿瘤的分型、分期、转移均无关(P>0.05);在分化表达中与前二者相同.Caspase-3定量分析中,癌组与对照组、鳞癌与腺癌组、高中低分化组、淋巴结转移与无淋巴结转移组之间均有不同程度的统计学意义.结论:Survivin、Pgp高表达时对肿瘤的原发耐药增强,结合Caspase-3定量分析,FI值均有不同程度的衰减.通过生物学行为研究,了解凋亡抑制是肿瘤耐药的主要原因,这有助于临床正确制订初始化疗方案和评估预后.

  13. Survivin gene silencing sensitizes prostate cancer cells to selenium growth inhibition

    Directory of Open Access Journals (Sweden)

    Liu Xichun

    2010-08-01

    Full Text Available Abstract Background Prostate cancer is a leading cause of cancer-related death in men worldwide. Survivin is a member of the inhibitor of apoptosis (IAP protein family that is expressed in the majority of human tumors including prostate cancer, but is barely detectable in terminally differentiated normal cells. Downregulation of survivin could sensitize prostate cancer cells to chemotherapeutic agents in vitro and in vivo. Selenium is an essential trace element. Several studies have shown that selenium compounds inhibit the growth of prostate cancer cells. The objective of this study is to investigate whether survivin gene silencing in conjunction with selenium treatment could enhance the therapeutic efficacy for prostate cancer and to elucidate the underlying mechanisms. Methods Expression of survivin was analyzed in a collection of normal and malignant prostatic tissues by immunohistochemical staining. In vitro studies were conducted in PC-3M, C4-2B, and 22Rv1 prostate cancer cells. The effect of selenium on survivin expression was analyzed by Western blotting and semi-quantitative RT-PCR. Survivin gene knockdown was carried out by transfecting cells with a short hairpin RNA (shRNA designed against survivin. Cell proliferation was quantitated by the 3-(4,5-Dimethylthiazol-2-yl- 2,5-Diphenyltetrazolium Bromide (MTT assay and apoptosis by propidium iodide staining followed by flow cytometry analysis. Finally, in vivo tumor growth assay was performed by establishing PC-3M xenograft in nude mice and monitoring tumor growth following transfection and treatment. Results We found that survivin was undetectable in normal prostatic tissues but was highly expressed in prostate cancers. Survivin knockdown or selenium treatment inhibited the growth of prostate cancer cells, but the selenium effect was modest. In contrast to what have been observed in other cell lines, selenium treatment had little or no effect on survivin expression in several androgen

  14. Survivin、RAGE和HMGB1在乳腺癌中表达及其临床意义%Expression of Survivin, RAGE and HMGB1 gene in human breast cancer and its clinical significance

    Institute of Scientific and Technical Information of China (English)

    鲁凯; 姚壮凯; 刘燕文; 叶红玲; 吕三云

    2012-01-01

    目的 探讨乳腺癌组织中Survivin、RAGE和HMGB1基因表达及其临床意义.方法 对50例早期(Ⅰ+Ⅱ期)、50例晚期(Ⅲ+Ⅳ期)乳腺癌及100例对照组蜡块标本,运用Real-time PCR和荧光原位杂交技术(FISH)技术检测Survivin、RAGE和HMGB1基因表达.并分析各基因表达与乳腺癌组织的分化程度、浸润深度、淋巴结转移、TNM分期之间的关系.结果 Survivin、RAGE和HMGB1基因表达荧光实时定量PCR法上调分别为62%、73%、79%,FISH法基因扩增阳性分别为78%、69%、72%.乳腺癌TNM分期、淋巴结转移与基因高表达有密切关系,Survivin基因表达与RAGE、HMGB1表达呈正相关.结论 Survivin、RAGE和HMGB1基因表达对乳腺癌早期诊断和预后分析有重要指导意义.%Objective To investigate the expression of Survivin,receptor for advanced glycation endproduct(RAGE) and high mobility group protein B1 (HMGB1) gene in human breast cancer and their clinical significance.Methods The expression of Survivin,RAGE and HMGB1 gene was detected by Real-time PCR technology and fluorescence in situ hybridization (FISH) technology in the following tissue samples:50 early breast cancers,50 advanced breast cancers,and the corresponding adjacent normal mammary tissues.The relationship of their expression and several factors such as differentiation degree,invasion,lymph node metastasis and TNM stage of cancer was explored.Results The positive expression rate of Survivin,RAGE and HMGB1 gene was 62%,73% and 79% detected by Real-time PCR technology,78%,69% and 72% detected by FISH technology in breast cancer tissues.The overexpression of these genes was positively correlated with TNM stage and lymph node metastasis.The expression of Survivin gene was positively correlated with the expression of RAGE and HMGB1.Conclusion Overexpression of Survivin,RAGE and HMGB1 gene is of great significance in early diagnosis and prognosis of human breast cancer.

  15. 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.

  16. Meta-analysis of the expression and significance of survivin in laryngeal carcinoma%生存素蛋白在喉癌组织中的表达和意义的meta分析

    Institute of Scientific and Technical Information of China (English)

    庞宇峰; 龚静蓉

    2012-01-01

    Objective To investigate the expression and significance of survivin in laryngeal carcinoma tissue.Methods Documents published in China from 2000 to 2011 were retrieved through CNKI,VIP,and wanfang database.The inclusive criteria of studies must be the case-control ones associated with survivin and laryngeal neoplasms.Taking into account the possibilities of heterogeneity of the studies,a statistical test for heterogeneity was performed.The Meta-analysis was applied with RevMan 4.3software.Results Twelve of case-control studies were selected for this Meta-analysis with a total of 605laryngeal carcinoma cases and 372 controls included.The pooled odds ratio between laryngeal carcinoma cases and normal membrana mucosa laryngis cases was 26.38,and 95% CI was ( 14.97,46.47 ).The pooled odds ratio between laryngeal carcinoma cases and peficancerous tissues cases was 4.19,and 95%CI was (2.50,7.03).The expression of Survivin was correlated with clinical stages (P < 0.01 ),pathological differentiation (P < 0.01) and lymph node metastasis (P < 0.05),but was not correlated with anatomical regions (P > 0.05 ).Conclusions Survivin may play an important role in the carcinogenesis,progress and metastasis of laryngeal carcinoma,which may be valuable for the diagnosis and prognosis.%目的 研究生存素( Survivin)蛋白在喉癌组织中的表达及意义.方法 以“Survivin”和“喉癌”为关键词在万方数据库、中国学术期刊数据库、中文科技期刊数据库上联合检索2000年至2011年国内专业期刊发表的关于Survivin在喉癌组织中表达的独立病例对照研究的论文,并对文献进行异质性检验,以病例组与对照组比值比(OR值)为效应指标,然后应用Meta分析软件RevMan4.3对各研究原始数据进行统计处理,并计算合并OR值及95%可信区间(95%CI),同时绘制Meta分析森林图.结果 有12篇文献纳入Meta分析,累计喉癌患者605例,对照组373例.喉癌

  17. Construction and identification of replication-competent adenovirus expressing siRNA targeting CD133 gene regulated by survivin promoter and its inhibition of liver cancer cell growth%survivin 启动子调控肿瘤干细胞标记 CD133基因 siRNA增殖型溶瘤腺病毒的构建及对肝癌细胞生长的抑制作用

    Institute of Scientific and Technical Information of China (English)

    牛坚; 王月; 刘斌; 王人颢; 朱志军; 申海莲

    2016-01-01

    目的:构建 survivin 启动子调控的靶向 CD133基因的 siRNA 增殖型溶瘤腺病毒,研究其对肝癌细胞生长的影响。方法RT-PCR 法扩增 survivin 启动子,测序鉴定,双酶切连接,获得 pH-XC2-survivin。酶切 pH-XC2-survivin、pZD55-CD133-siRNA 获得 survivin 启动子表达框的亚克隆和CD133-siRNA 基因表达框的亚克隆,连接获得 survivin 启动子调控的 siRNA 增殖型溶瘤腺病毒表达载体质粒 pT-ZD55-CD133-siRNA。增殖型溶瘤腺病毒 survivin-T-ZD55-CD133-siRNA 经 PCR 和测序鉴定。 qRT-PCR 法检测 CD133表达, Western blot 法检测 E1A,CCK-8法检测细胞生长,流式细胞术检测细胞凋亡。结果成功构建增殖型溶瘤腺病毒 sur-vivin-T-ZD55-CD133-siRNA。 qRT-PCR 法检测 CD133 mRNA明显下降, Western blot 证实 survivin-T-ZD55-CD133-siRNA在肿瘤细胞中表达 E1A 能抑制肝癌细胞 CD133表达及生长。结论构建的增殖型溶瘤腺病毒可有效降低肝癌细胞CD133的表达,用于肝癌基因治疗的进一步研究。%Objective To construct a replication-competent adenovirus expressing siRNA targeting CD133 gene regulated by survivin promoter and investigate its inhibitory effect on Hep 3B cells.Methods The fragment of the survivin promoter was amplified by PCR and inserted into pH -XC2 to reconstruct a recombinant plasmid pH -XC2-survivin.Complete digestion pH-XC2-survivin and pZD55-CD133-siRNA, combinational joining the subclones, then getting replication-competent adenovirus expressing short interference RNA targeting CD 133 gene regulated by survivin promoter, replication-competent adenovirus was constructed .The recombined adenoviruses ( T-ZD55-CD133-siRNA) were verified by PCR and sequencing .The effect of T-ZD55-CD133-siRNA on CD133 expression in Hep3B cells was detected by qRT-PCR.The expression of E1A was detected by Western blot.The antitumor po-tential of replication

  18. Survivin minigene DNA vaccination is effective against neuroblastoma.

    Science.gov (United States)

    Fest, Stefan; Huebener, Nicole; Bleeke, Matthias; Durmus, Tahir; Stermann, Alexander; Woehler, Anja; Baykan, Bianca; Zenclussen, Ana C; Michalsky, Elke; Jaeger, Ines S; Preissner, Robert; Hohn, Oliver; Weixler, Silke; Gaedicke, Gerhard; Lode, Holger N

    2009-07-01

    The inhibitor of apoptosis protein survivin is highly expressed in neuroblastoma (NB) and survivin-specific T cells were identified in Stage 4 patients. Therefore, we generated a novel survivin minigene DNA vaccine (pUS-high) encoding exclusively for survivin-derived peptides with superior MHC class I (H2-K(k)) binding affinities and tested its efficacy to suppress tumor growth and metastases in a syngeneic NB mouse model. Vaccination was performed by oral gavage of attenuated Salmonella typhimurium SL7207 carrying pUS-high. Mice receiving the pUS-high in the prophylactic setting presented a 48-52% reduction in s.c. tumor volume, weight and liver metastasis level in contrast to empty vector controls. This response was as effective as a survivin full-length vaccine and was associated with an increased target cell lysis, increased presence of CD8(+) T-cells at the primary tumor site and enhanced production of proinflammatory cytokines by systemic CD8(+) T cells. Furthermore, depletion of CD8(+) but not CD4(+) T-cells completely abrogated the pUS-high mediated primary tumor growth suppression, demonstrating a CD8(+) T-cell mediated effect. Therapeutic vaccination with pUS-high led to complete NB eradication in over 50% of immunized mice and surviving mice showed an over 80% reduction in primary tumor growth upon rechallenge in contrast to controls. In summary, survivin-based DNA vaccination is effective against NB and the rational minigene design provides a promising approach to circumvent potentially hazardous effects of using full length antiapoptotic genes as DNA vaccines.

  19. RNAi抑制存活素基因表达对前列腺癌细胞侵袭的影响%Effects of RNA interference-mediated silencing expression of survivin gene on invasion of human prostate cancer cell

    Institute of Scientific and Technical Information of China (English)

    范钰; 张尤历; 李华

    2008-01-01

    1.4, and the control group was 22.6±1.8(P<0.05). The results from boyden chamber model exhibited that the cells numbers crossing filter membrane in group of 3.125,6.25 and 12.5 nmol/L of siRNA were 33.6±2.1,19.5±1.9,8.1±1.83, and the control group was 49.4±2.3(all P<0.05). The results in vivo showed that cancer cells of control groups invaded into striped muscle and blood vessel, and there were no these phenomenons in transferred group with survivin siRNA. Survivin siRNA could reduce expression level of MMP-2 and MMP-9 in prostate cancer cells(P<0. 01). Conclusions Survivin-directed RNA interference can inhibit invasion of human prostate cancer cell through down-regualting MMP-2,-9 genes.

  20. Detection of Apoptotic Inhibitor Gene Survivin in Peripheral Blood of Patients with Esophageal Cancer by Real-time Fluorescence Quantitative PCR and its Clinical Signiifcance

    Institute of Scientific and Technical Information of China (English)

    CHEN Sheng

    2014-01-01

    Objective:To explore the clinical signiifcance of apoptotic inhibitor gene Survivin in peripheral blood of patients with esophageal cancer. Methods:Real-time lfuorescence quantitative PCR was used to detect the expression of Survivin mRNA in peripheral blood of 93 patients with benign and malignant esophageal lesions. The relationship of Survivin mRNA expression and clinicopathologic feature was observed. Results:The expression of Survivin mRNA in peripheral blood which was associated with differentiated degree and clinical staging was progressively increased from benign lesion to carcinoma in situ and invasive carcinoma. Conclusion:The expression of Survivin mRNA in peripheral blood is significantly related to the genesis and progression of esophageal carcinoma. Real-time fluorescence quantitative PCR used to detect the expression of Survivin m-RNA in peripheral blood may be more convenient for diagnosing and guiding the treatment of esophageal carcinoma.

  1. Survivin as a potential mediator to support autoreactive cell survival in myasthenia gravis: a human and animal model study.

    Directory of Open Access Journals (Sweden)

    Linda L Kusner

    Full Text Available The mechanisms that underlie the development and maintenance of autoimmunity in myasthenia gravis are poorly understood. In this investigation, we evaluate the role of survivin, a member of the inhibitor of apoptosis protein family, in humans and in two animal models. We identified survivin expression in cells with B lymphocyte and plasma cells markers, and in the thymuses of patients with myasthenia gravis. A portion of survivin-expressing cells specifically bound a peptide derived from the alpha subunit of acetylcholine receptor indicating that they recognize the peptide. Thymuses of patients with myasthenia gravis had large numbers of survivin-positive cells with fewer cells in the thymuses of corticosteroid-treated patients. Application of a survivin vaccination strategy in mouse and rat models of myasthenia gravis demonstrated improved motor assessment, a reduction in acetylcholine receptor specific autoantibodies, and a retention of acetylcholine receptor at the neuromuscular junction, associated with marked reduction of survivin-expressing circulating CD20+ cells. These data strongly suggest that survivin expression in cells with lymphocyte and plasma cell markers occurs in patients with myasthenia gravis and in two animal models of myasthenia gravis. Survivin expression may be part of a mechanism that inhibits the apoptosis of autoreactive B cells in myasthenia gravis and other autoimmune disorders.

  2. Survivin as a potential mediator to support autoreactive cell survival in myasthenia gravis: a human and animal model study.

    Science.gov (United States)

    Kusner, Linda L; Ciesielski, Michael J; Marx, Alexander; Kaminski, Henry J; Fenstermaker, Robert A

    2014-01-01

    The mechanisms that underlie the development and maintenance of autoimmunity in myasthenia gravis are poorly understood. In this investigation, we evaluate the role of survivin, a member of the inhibitor of apoptosis protein family, in humans and in two animal models. We identified survivin expression in cells with B lymphocyte and plasma cells markers, and in the thymuses of patients with myasthenia gravis. A portion of survivin-expressing cells specifically bound a peptide derived from the alpha subunit of acetylcholine receptor indicating that they recognize the peptide. Thymuses of patients with myasthenia gravis had large numbers of survivin-positive cells with fewer cells in the thymuses of corticosteroid-treated patients. Application of a survivin vaccination strategy in mouse and rat models of myasthenia gravis demonstrated improved motor assessment, a reduction in acetylcholine receptor specific autoantibodies, and a retention of acetylcholine receptor at the neuromuscular junction, associated with marked reduction of survivin-expressing circulating CD20+ cells. These data strongly suggest that survivin expression in cells with lymphocyte and plasma cell markers occurs in patients with myasthenia gravis and in two animal models of myasthenia gravis. Survivin expression may be part of a mechanism that inhibits the apoptosis of autoreactive B cells in myasthenia gravis and other autoimmune disorders.

  3. Survivin Overexpression Is Associated with Aggressive Clinicopathological Features in Cervical Carcinoma: A Meta-Analysis

    Science.gov (United States)

    Cheng, Ke-yan; Wang, Zhi-lian; Gu, Qian-yun; Hao, Min

    2016-01-01

    Objective Overexpression of survivin has been reported in many human tumors. However, the clinicopathological features associated with survivin overexpression in cervical carcinoma remain controversial. Thus, the current meta-analysis was performed to assess the clinicopathological significance of survivin in cervical carcinoma. Methods PubMed, EMBASE, and Web of Science databases were searched for relevant studies published through November 1, 2015. A meta-analysis was performed to evaluate the association between survivin expression and clinicopathological outcome in cervical carcinoma. Results Eleven eligible studies with a total of 865 patients were included. Survivin overexpression was closely related to lymph node metastasis (odds ratio [OR] = 0.679, 95% confidence interval [CI]: 0.509–0.905, P = 0.008) but was not significantly associated with tumor FIGO stage (I+II vs. III+IV) (OR = 0.843, 95% CI: 0.626–1.137, P = 0.264), tumor grade (G1+G2 vs. G3) (OR = 0.913, 95% CI: 0.689–1.210, P = 0.527), tumor size (>4 vs. ≤4 cm) (OR = 0.825, 95% CI: 0.434–1.570, P = 0.559), or stromal involvement (OR = 0.820, 95% CI: 0.545–1.233, P = 0.340). The correlation between survivin expression and overall survival was evaluated among a total of 238 patients from three eligible studies. The pooled HR was 1.129 (95% CI: 0.597–1.661; P = 0.000), indicating that survivin expression was significantly associated with poor survival in cervical carcinoma. Conclusions Based on the current meta-analysis, survivin is strongly associated with lymph node metastasis and poor prognosis. Additionally, survivin is a novel clinicopathological marker of cervical carcinoma and thus may be a therapeutic target for cervical carcinoma. PMID:27764228

  4. Downregulation of survivin by siRNA inhibits invasion and promotes apoptosis in neuroblastoma SH-SY5Y cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, L.; Liang, H. [Department of Pediatrics, Qilu Hospital, Shandong University, Jinan (China); Cao, W. [Department of Obstetrics, Qingdao Central Hospital, Qingdao (China); Xu, R.; Ju, X.L. [Department of Pediatrics, Qilu Hospital, Shandong University, Jinan (China)

    2014-05-23

    Neuroblastoma is a solid tumor that occurs mainly in children. Malignant neuroblastomas have a poor prognosis because conventional chemotherapeutic agents are not very effective. Survivin, a member of the inhibitor of the apoptosis protein family, plays a significant role in cell division, inhibition of apoptosis, and promotion of cell proliferation and invasion. Previous studies found that survivin is highly expressed in some malignant neuroblastomas and is correlated with poor prognosis. The aim of this study was to investigate whether survivin could serve as a potential therapeutic target of human neuroblastoma. We employed RNA interference to reduce survivin expression in the human neuroblastoma SH-SY5Y cell line and analyzed the effect of RNA interference on cell proliferation and invasion in vitro and in vivo. RNA interference of survivin led to a significant decrease in invasiveness and proliferation and increased apoptosis in SH-SY5Y cells in vitro. RNA interference of survivin inhibited tumor growth in vivo by 68±13% (P=0.002) and increased the number of apoptotic cells by 9.8±1.2% (P=0.001) compared with negative small interfering RNA (siRNA) treatment controls. Moreover, RNA interference of survivin inhibited the formation of lung metastases by 92% (P=0.002) and reduced microvascular density by 60% (P=0.0003). Survivin siRNA resulted in significant downregulation of survivin mRNA and protein expression both in vitro and in vivo compared with negative siRNA treatment controls. RNA interference of survivin was found to be a potent inhibitor of SH-SY5Y tumor growth and metastasis formation. These results support further clinical development of RNA interference of survivin as a treatment of neuroblastoma and other cancer types.

  5. The inhibitory effect of Sulindac on human pancreatic cancer cells' proliferation by targeting survivin/ Aurora B pathway%舒林酸经survivin/Aurora B途径对人胰腺癌细胞分裂的阻断效应

    Institute of Scientific and Technical Information of China (English)

    范学科; 廖宇圣; 张翠芳; 陈芬; 高慧涛; 覃华; 李德民; 赵秋

    2008-01-01

    Objective To observe the expression of survivin and Aurora B in human pancreatic cancer BXPC3 cells after the treatment of sulindac and to explore the potential mechanism. Methods MTr assay was used to determine the effect of sulindac on the proliferation of the BXPC3 cells. RT-PCR was used to detect the expression of mRNA level of survivin and Aurora B, western blot was used to detect protein expression of survivin and Aurora B Thr-232. Cell cycle and apoptosis were detected by flow eytometry (FCM). Results The BXPC3 cells were inhibited by sulindac in a dose and time-dependent manner; the expression of mRNA of survivin and Aurora B were both significantly decreased from 1.5644 and 0.6554 to 0. 4372 and 0.1132 (P< 0.01), the expression of survivin protein and the phosphorylation of Aurora B Thr-232 were also decreased from 1.2735 and 0.4680 to 0.2126 and 0.2546 (P<0.01); the proportion of cells in the G0/G1 phase was increased from (56.65±1.93)% to (70.58±3.21)% (P<0.01). Conclusions Sulindac had inhibitory effects on the growth of BXPC3 cells, the possible mechanism was via decreasing the expression of survivin which depressed the activity of Aurora B, then the CPC was influenced. The most of the cells were blocked in the G0/G1 phase, and the cells' mitosis was inhibited.%目的 观察舒林酸处理胰腺癌细胞BxPC3后对survivin、Aurora B表达及细胞周期和增殖的影响,探讨舒林酸的作用机制.方法 应用MTT法检测舒林酸对BxPC3细胞的增殖抑制作用,RT-PCR法检测survivin mRNA、Aurora B mRNA的表达,Western blot法检测survivin蛋白表达及Aurora BThr-232磷酸化水平,流式细胞仪检测细胞周期变化.结果 舒林酸呈时间和剂量依赖性抑制BxPC3细胞增殖.经500μmoL/L舒林酸作用细胞48 h后,survivin mRNA和Aurora B mRNA表达量分别从1.56和0.66下降到0.44和0.11(P<0.01);survivin蛋白表达从1.27下降到0.21(P<0.01),Aurora BThr-232磷酸化水平从0.47下降到0.25(P<0.01);G0/G1

  6. Expression of survivin gene in animal model of posterior capsule opacification in rats%Survivin基因在大鼠后发性白内障动物模型中的表达

    Institute of Scientific and Technical Information of China (English)

    陈颖; 陆斌; 吴强; 张敏

    2011-01-01

    目的 建立SD大鼠后发性白内障(PCO)动物模型,并检测Survivin调控基因在PCO中的表达变化,探讨Survivin对PCO生成的影响.方法 取成年SD大鼠60只随机分为6组,其中10只(n=10)作为正常对照组,另50只SD大鼠于腹腔麻醉联合表面麻醉下行右眼晶状体囊外摘出术(ECLE),分为A、B、C、D、E,每10只(n=10)为1组,分别于术后即刻、3、7、14、28 d对术眼进行裂隙灯显微镜及组织病理学检查,观察PCO形成的时间、部位、发展过程及组织形态学改变,并采用逆转录聚合酶链反应(RT-PCR)方法及免疫组化方法检测Survivin基因在术后不同时间点PCO中的表达.结果 PCO在术后3 d出现,可见后囊膜皱褶,术后7、14 d后囊膜混浊明显,所有动物于术后14 d出现明显的后囊膜皱缩,可见新生晶状体纤维,术后28 d可见明显的后囊膜增厚,新生晶状体纤维填充囊袋,并且透明度下降.免疫组化和RT-PCR于正常对照组及术后即刻A组均未检测到Survivin的表达,B、C、D、E4组术后不同时段的PCO组织中可检测到不同程度Survivin的表达,其中,免疫组化显示C、D 2组表达较为明显,RT-PCR则显示C组达到表达高峰,D组表达开始下降,但较E组表达强,B组表达最弱.结论 SD大鼠可成功建立PCO动物模型并检测到Survivin调控基因的表达,Survivin的表达在PCO的形成过程中具有相应的时相性,提示 Survivin与PCO的形成机制具有一定相关性,对探索PCO的基因治疗方法具有参考价值.%Objective This study is to establish animal models of posterior capsule opacification ( PCO ) in Spra-gue dawley(SD ) rats and detect the expression of the survivin in PCO tissue. Methods 60 adult SD rats were divided randomly into six groups: 10( n = 10 ) in control group; extra capsular lens extraction ( ECLE ) were performed in the other 50 rats under abdominal cavity anesthesia combined with topical anesthesia . The operative eyes were divided into A

  7. Effect of protein kinase C alpha, caspase-3, and survivin on apoptosis of oral cancer cells induced by staurosporine

    Institute of Scientific and Technical Information of China (English)

    Yu-xia ZHANG; Shi-bin YU; Jing-ping OU-YANG; Dong XIA; Min WANG; Jin-rong LI

    2005-01-01

    Aim: To elucidate inhibition of protein kinase C α (PKC α) activity by staurosporine on apoptosis of oral cancer cell line tongue squamous cell carcinoma (TSCCa)cells and to clarify the role of survivin and caspase-3 in mediating apoptosis.Methods: TSCCa cell viability was measured by MTT assay after 100 nmol/L staurosporine treatment. Apoptotic cells were identified by using phase contrast microscopy, acridine orange/ethidium bromide staining, and flow cytometry. Level of PKC α and its subcellular location were investigated using Western blot analysis.Expression of survivin and caspase-3 were evaluated using immunocytochemistry.Results: Staurosporine significantly inhibited the cell viability of TSCCa cells in a dose- and time-dependent manner. Marked cell accumulation in G2/M phase was observed after 100 nmol/L staurosporine exposure for 6 h and 12 h. In addition,the percentage of apoptosis increased in a time-dependent manner, from 2.9% in control cultures to approximately 27.4% at 100 nmol/L staurosporine treatment for24 h. Staurosporine displayed difference in inhibitory efficacy between cytosolic and membrance-derived PKC α. The content of PKCα in membrane versus cytosol decreased quickly, from 0.45 in ethanol-treated control cultures to 0.18 after staurosporine exposure for 24 h (P<0.01). After treatment withstaurosporine, a time-dependent reduction of survivin and an activation of caspase-3 were observed in TSCCa cells. Conclusion: Staurosporine inhibited cell viability and promoted apoptosis in TSCCa cells. Inhibition of PKCα activity might be a potential mechanism for staurosporine to induce apoptosis in this cell line. The cleavage of survivin and activation of caspase-3 signaling pathway might contribute to PKC α inhibition-induced apoptosis.

  8. Experimental cancer gene therapy by multiple anti-survivin hammerhead ribozymes

    Institute of Scientific and Technical Information of China (English)

    Qi Fei; Yuwen Ke; Xuebiao Yao; Jingde Zhu; Hongyu Zhang; Lili Fu; Xinlan Dai; Baomei Gao; Min Ni; Chao Ge; Jinjun Li; Xia Ding

    2008-01-01

    To improve the efficacy of gene therapy for cancer, we designed four hammerhead ribozyme adenoviruses (R1 to R4) targeting the exposed regions of survivin mRNA. In addition to the in vitro characterization, which included a determination of the sequence specificity of cleavage by primer extension, assays for cell proliferation and for in vivo tumor growth were used to score for ribozyme efficiency.The resulting suppression of survivin expression induced mitotic catastrophe and cell death via the caspase-3-dependent pathway. Importantly, administration of the ribozyme adenoviruses inhibited tumor growth in a hepato-cellular carcinoma xenograft mouse model. Co-expression of R1, R3 and R4 ribozymes synergistically suppressed survivin and, as this combination targets all major forms of the survivin transcripts, produced the most potent anti-cancer effects. The adenoviruses carrying the multiple hammerhead ribozymes described in this report offered a robust gene therapy strategy against cancer.

  9. Nanoformulated cell-penetrating survivin mutant and its dual actions

    Directory of Open Access Journals (Sweden)

    Sriramoju B

    2014-07-01

    Full Text Available Bhasker Sriramoju, Rupinder K Kanwar, Jagat R Kanwar Nanomedicine Laboratory of Immunology and Molecular Biomedical Research (NLIMBR, School of Medicine, Faculty of Health, Deakin University, Geelong, Australia Abstract: In this study, we investigated the differential actions of a dominant-negative survivin mutant (SurR9-C84A against cancerous SK-N-SH neuroblastoma cell lines and differentiated SK-N-SH neurons. In both the cases, the mutant protein displayed dual actions, where its effects were cytotoxic toward cancerous cells and proliferative toward the differentiated neurons. This can be explained by the fact that tumorous (undifferentiated SK-N-SH cells have a high endogenous survivin pool and upon treatment with mutant SuR9-C84A causes forceful survivin expression. These events significantly lowered the microtubule dynamics and stability, eventually leading to apoptosis. In the case of differentiated SK-N-SH neurons that express negligible levels of wild-type survivin, the mutant indistinguishably behaved in a wild-type fashion. It also favored cell-cycle progression, forming the chromosome-passenger complex, and stabilized the microtubule-organizing center. Therefore, mutant SurR9-C84A represents a novel therapeutic with its dual actions (cytotoxic toward tumor cells and protective and proliferative toward neuronal cells, and hence finds potential applications against a variety of neurological disorders. In this study, we also developed a novel poly(lactic-co-glycolic acid nanoparticulate formulation to surmount the hurdles associated with the delivery of SurR9-C84A, thus enhancing its effective therapeutic outcome. Keywords: survivin mutant, neurological disorders, protein therapeutics, inhibitor of apoptosis protein family, poly(lactic-co-glycolic acid

  10. An indirubin derivative, indirubin-3'-monoxime suppresses oral cancer tumorigenesis through the downregulation of survivin.

    Directory of Open Access Journals (Sweden)

    Wan-Yu Lo

    Full Text Available Oral cancer is the fourth most common cause of death from cancer in Taiwanese men. Indirubin-3'-monoxime (I3M, a potent cyclin-dependent kinase inhibitor, has therapeutic effects in other cancer cells. In this study, we carried out in vitro assays to test cell viability, cell cycle progression, apoptosis, cell migration and invasion in this cancer type. In addition, using an oral tumorigenic animal model, we examined target gene and protein expression using real time qPCR, immunoblotting and immunohistochemical staining. Our results demonstrate that I3M has an anti-proliferative effect in both Cal-27 and HSC-3 oral cancer cell lines and that treatment of Cal-27 and HSC-3 cells with I3M results in apoptosis through the activation of cytochrome c. In addition, I3M interrupts the cell cycle in Cal-27 cells in a dose-dependent manner by arresting cells in the G2/M phase. We also found that I3M suppresses migration and invasion in Cal-27 cells by inhibiting the expression of focal adhesion kinase, urokinase-type plasminogen inhibitor, and matrix metalloproteinase 9. Moreover, we identified survivin as a target protein in I3M-treated oral cancer cells. Using an oral cancer mouse model, we demonstrate that topical application of an adhesive gel composed of I3M and poly(vinyl alcohol (I3M/PVA has dose-dependent anti-tumorigenic effects. Following treatment, the expression of survivin protein and mRNA was downregulated in cancerous tissues. Furthermore, plasma survivin levels were also reduced in the I3M-treated mice. These results suggest that topical application of I3M, a drug synthesized from indirubin, which is found in Qing-Dai - has therapeutic potential for treating oral cancer.

  11. Survivin ASODN targeted therapy in XWLC-05 cell transplanted nude mice

    Institute of Scientific and Technical Information of China (English)

    Weiwei Wang; Shaojia Wang; Gaofeng Li; Lei Li; Ruibing Cheng

    2012-01-01

    Objective: The aim of this study was to study the inhibiting effect of survivin mRNA on transplanted XWLC-05 tumor on nude mice. Methods: We established XWLC-05 transplanted nude mice model. 44 mice would be divided randomly into 4 groups: control group (blank), Lip group (simple liposome), survivin SODN group (transfected by sense oligonudeotide) and survivin ASODN group (transfected by antisense oligonudeotide). We would study general activities of nude mice in these 4 groups, measure the size of tumor and calculate the tumor inhabiting rate also. Pathological methods were applied in the analysis of the effect of different treatment on heart, kidney and liver of nude mice in these 4 groups. Results: Tumor grew slowly and size, weight of tumor was lower in survivin ASODN group when compared with that of others. Nude mice of survivin ASODN group showed lower growth index and tumor inhabiting rate was significantly higher than that of other groups (P 0.05). We found a great deal of tumor cell necrosis in survivin ASODN group. No death of nude mice was observed in all 4 groups and we did not found obvious lesion in vital organs. Conclusion: Survivin ASDON could be used for the inhibition of subcutaneously transplanted tumor in nude mice without obvious lesion in vital organs.

  12. 凋亡相关因子 Survivin 和 Smac 在卵巢癌中的研究进展%Research progress of ovarian cancer apoptosis related factor-Survivin and Smac

    Institute of Scientific and Technical Information of China (English)

    马丹; 马玲

    2015-01-01

    Survivin is the strongest member of the inhibitor of apoptosis protein(IAP)family. Smac is the second mitochondria-derived activator of cysteine proteases,which can promote apoptosis by combining with IAP. Abnormal expression of them is closely related with occurrence,development,treatment tolerance and prognosis of ovarian cancer. It is prompted that Survivin and Smac are expected to play important roles in the early diagnosis and targeted therapy of ovarian cancer.%Survivin 是凋亡抑制蛋白(IAP)家族中抗凋亡作用最强的成员。Smac 是半胱氨酸蛋白酶的第2个线粒体激活因子,可以通过与 IAP 的结合,来发挥促凋亡作用。二者在卵巢癌组织中的异常表达均与卵巢癌发生发展、治疗耐受及预后密切相关。两者有望在卵巢癌早期诊断及靶向治疗方面发挥重要作用。

  13. Survivin启动子调控的PUMA基因表达载体对乳腺癌MCF-7细胞凋亡的影响%Construction of a PUMA gene-targeted expression vector regulated by Survivin promoter and research of its effect on cell apoptosis in MCF-7 cells

    Institute of Scientific and Technical Information of China (English)

    陈绍坤; 刘晓华; 黄丽; 余红; 税青林

    2013-01-01

    目的:探讨构建带有肿瘤特异性Survivin启动子的PUMA基因表达载体pUC57-Survivin-PUMA对乳腺癌MCF-7细胞的肿瘤特异性促凋亡作用.方法:化学合成PUMA基因片段克隆至pUC57质粒,构建重组质粒pUC57-PUMA;克隆Survivin启动子序列,取代pUC57-PUMA质粒原有的CMV启动子,构建重组质粒pUC57-Survivin-PUMA,测序鉴定.实验设置pUC57-PUMA组、pUC57-Survivin-PUMA组、阴性对照组(pUC57group)和空白对照组(blank group).将以上各组分别转染人乳腺癌MCF-7细胞及人正常乳腺Hs 578Bst细胞.转染后48h,western blot技术检测各组细胞中PUMA蛋白表达情况,流式细胞仪检测细胞的凋亡率.结果:重组质粒经测序鉴定证实符合设计要求,构建成功;MCF-7细胞中pUC57-PUMA组和pUC57-Survivin-PUMA组的PUMA蛋白表达均较对照组显著增高(P<0.05),而人正常乳腺Hs 578Bst细胞中只有pUC57-PUMA组的PUMA蛋白表达较其他3组显著增高(P<0.05).MCF-7细胞中pUC57-PUMA组和pUC57-Survivin-PUMA组的细胞凋亡率分别为29.02%和29.31%,均较对照组显著增高(P<0.05),而人正常乳腺Hs 578Bst细胞中仅pUC57-PUMA组的细胞凋亡率较其他3组显著增高(P<0.05).结论:Survivin启动子调控的PUMA表达载体能显著增高乳腺癌MCF-7细胞中PUMA的表达,进而促进乳腺癌MCF-7细胞凋亡,而对正常乳腺Hs 578Bst细胞的凋亡不产生影响.%Objective:To construct PUMA gene-targeted expression vector driven by tumor specific Survivin promoter (pUC57-Survivin-PUMA) and investigate its cell apoptosis effects in MCF-7 breast cancer cells.Methods:The PUMA gene sequence by chemical synthesis was cloned in the plasmid pUC57 to construct the recombinant plasmid pUC57-PUMA,the CMV promoter in which then were replaced respectively by the Survivin promoter,and the plasmid pUC57-Survivin-PUMA was constructed finally.Four experimental groups were set up,they were pUC57-PUMA group,pUC57-Survivin-PUMA group,pUC57 control group

  14. Clinical significance of survivin in the diagnosis and prognosis of endometrial carcinoma

    Institute of Scientific and Technical Information of China (English)

    Yanxiang Cheng; Gantao Chen; Yanjun Cheng; Demin Pu

    2006-01-01

    Objective: To investigate the clinical significance of survivin in endometrial carcinoma and to investigate the relationship between the expression of survivin and Ki-67. Methods: Immunohistochemical S-P (streptavidin-biotin-peroxidase complex)method was performed to detect the expression of survivin and Ki-67 antigen in 15 cases of normal endometrium, 21 cases of endometrial simple and complex hyperplasia, 22 cases of endometrial atypical hyperplasia, and 61 cases of endometrial carcinoma. Results: Survivin was hardly detected in some normal endometrium in the proliferative phase and in the secretory phase. However, the level of survivin expression in atypical hyperplasia endometrium(72.73%)was higher than that in normal endometrium (7.14%)(P < 0.05), including simple and complex hyperplasia (42.38%)(P < 0.01), and was lower than that in endometrial carcinoma (90.17%)(P < 0.05). Moreover, significant correlation was present between the expression of survivin and the characteristics of endometrial carcinoma, including clinical stage, histological grade and the presence of invasion to myometrium (P < 0.05). In addition, Ki-67 antigen expression was positively correlated with survivin expression in all specimen. Ki-67 labeled indexes (LIs)in hyperplasia endometrium were significantly lower than those in atypical hyperplasia endometrium and endometrial carcinoma (P < 0.01 ), while there was no significant difference in Ki-67 LIs between atypical hyperplasia endometrium and endometrial carcinoma(P > 0.05). There was no significant relationship between Ki-67 LIs and the characteristics of endometrial carcinoma, including histological grade, clinical stage or the invasion to myometrium(P > 0.05). Conclusion: Survivin may participate in the onset and progression of endometrial carcinoma through inhibiting apoptosis and promoting proliferation. Survivin expression is correlated with the malignant degree and prognosis of tumor. Ki-67 is also associated with

  15. Obstructive Effects of Ultrasonic Microbubble Intensifier on CHG-5 Cell with Survivin Antisense Oligonucleotides Transfection

    Institute of Scientific and Technical Information of China (English)

    CAO Hong-ying; CAO You-de; WANG Zhi-gang; LI Pan

    2008-01-01

    Objective:To study the effects on human glioma cell line CHG-5 by ultrasonic microbubble intensifier with survivin antisense oligonucleotides (ASODN)transfection. Methods: Antisense oligonucleotides targeting survivin mRNA was designed and synthesized.Four regimen groups were designed,group A:survivin antisense oligonucleotides transfected with ultrasonic microbubble intensifier combined with ultrasound irradiation,group B: survivin antisense oligonucleotides transfected with lipofectamine combined with ultrasound irradiation,group C:survivin antisense oligonucelotides with lipofectamine transfection.group D:blank control.The expression changes of surviving protein were measured by immunohistochemical staining and Western blotting,and MTr assay was used to measure the changes of proliferation.Results:Survivin protein expression in group A was decreased significantly in human glioma cell line CHG-5 than other groups(P<0.05),and the proliferating rate of CHG-5 in group A was also significantly inhibited(P<0.05).Conclusion:Ultrasonic microbubble intensifier transfection combined with ultrasound irradiation is a promising method in gene transfection effectively and noninvasively.

  16. Plasma-derived exosomal survivin, a plausible biomarker for early detection of prostate cancer.

    Directory of Open Access Journals (Sweden)

    Salma Khan

    Full Text Available BACKGROUND: Survivin is expressed in prostate cancer (PCa, and its downregulation sensitizes PCa cells to chemotherapeutic agents in vitro and in vivo. Small membrane-bound vesicles called exosomes, secreted from the endosomal membrane compartment, contain RNA and protein that they readily transport via exosome internalization into recipient cells. Recent progress has shown that tumor-derived exosomes play multiple roles in tumor growth and metastasis and may produce these functions via immune escape, tumor invasion and angiogenesis. Furthermore, exosome analysis may provide novel biomarkers to diagnose or monitor PCa treatment. METHODS: Exosomes were purified from the plasma and serum from 39 PCa patients, 20 BPH patients, 8 prostate cancer recurrent and 16 healthy controls using ultracentrifugation and their quantities and qualities were quantified and visualized from both the plasma and the purified exosomes using ELISA and Western blotting, respectively. RESULTS: Survivin was significantly increased in the tumor-derived samples, compared to those from BPH and controls with virtually no difference in the quantity of Survivin detected in exosomes collected from newly diagnosed patients exhibiting low (six or high (nine Gleason scores. Exosome Survivin levels were also higher in patients that had relapsed on chemotherapy compared to controls. CONCLUSIONS: These studies demonstrate that Survivin exists in plasma exosomes from both normal, BPH and PCa subjects. The relative amounts of exosomal Survivin in PCa plasma was significantly higher than in those with pre-inflammatory BPH and control plasma. This differential expression of exosomal Survivin was seen with both newly diagnosed and advanced PCa subjects with high or low-grade cancers. Analysis of plasma exosomal Survivin levels may offer a convenient tool for diagnosing or monitoring PCa and may, as it is elevated in low as well as high Gleason scored samples, be used for early detection.

  17. 喉癌生存素表达临床特征分析%Clinical expression of survivin in larynx cancer

    Institute of Scientific and Technical Information of China (English)

    唐梓轩; 张静; 杨立

    2008-01-01

    目的:探讨生存素(survivin)表达与喉癌发生、发展及浸润转移的关系.方法:喉癌组为65例肿瘤组织.均经病理确诊为喉鳞状细胞癌.采用免疫组织化学SABC染色法检测survivin蛋白表达情况,并进行统计分析.结果:喉癌组survivin蛋白定位于癌细胞胞浆,阳性率(51/65,78.5%)显著高于癌旁组(2/22,9.1%),淋巴转移组显著高于无转移组,不同临床分期阳性率显著不同.结论:喉癌组织中survivin表达水平较高,survivin表达与喉癌发生、发展及浸润转移有关,对喉癌的早期诊断、预后判断有意义.

  18. 实时荧光定量RT-PCR检测鼻咽癌Survivin mRNA基因表达%Detection of Survivin mRNA in nasopharyngeal carcinoma by real-time fluorescence quantitative RT-PCR

    Institute of Scientific and Technical Information of China (English)

    Shengmiao Fu; Junhong Cai; Zhihua Tu; Yutian Wang; Liqun Deng; Zhu Liang; Zhenqun Lin; Xuanju Gong

    2008-01-01

    Objective:To establish the method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in nasopharyngeal carcinoma (NPC) tissues.Methods:The total RNA was extracted from NPC cell line CNE-2 and tissues with Trizol and then been transcribed reversely to cDNA,a method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in NPC tissues had been established,in which chronic nasopharyngitis patients' nasopharynx tissues treated as control group.Results:The expression of Survivin mRNA all could be detected either in CNE-2 cells,NPC tissues or in chronic nasopharyngitis patients' nasopharynx tissues,and there was higher the expression level of Survivin mRNA in NPC tissues than which in chronic nasopharyngitis patients' nasopharynx tissues,the difference was significant (P<0.01).The expression of Survivin mRNA could be detected both in stage Ⅰ+Ⅱ and stage Ⅲ+Ⅳ NPC,and there was no significant difference in relative quantifications of gene expression between these two groups (P>0.05).There was no relationship between Survivin mRNA expression and age and sex of NPC patients (P>0.05).Conclusion:Real time fluorescence quantitative RT-PCR is a rapid,effective and high sensitive method for detecting the expression of Survivin mRNA in NPC tissues.The overexpression of Survivin mRNA may play some roles in pathogenesis of NPC.

  19. A polymorphism in the promoter region of the survivin gene is related to hemorrhagic transformation in patients with acute ischemic stroke.

    Science.gov (United States)

    Mallolas, Judith; Rodríguez, Rocío; Gubern, Carme; Camós, Susanna; Serena, Joaquín; Castellanos, Mar

    2014-12-01

    Hemorrhagic transformation (HT) of cerebral infarction is a common and serious occurrence following acute ischemic stroke. The expression of survivin, a member of the inhibitor of apoptosis protein family, has been shown to increase after cerebral ischemia. This protein has been mainly located at the microvasculature within the infarcted and peri-infarcted area, so we aimed to investigate whether survivin gene polymorphisms, also known as BIRC5 gene, were associated with HT of cerebral infarction. Polymorphism screening of the BIRC5 gene was performed in 107 patients with a hemispheric ischemic stroke and 93 controls by polymerase chain reaction, single-strand conformation polymorphism and sequencing analysis. Genotype-phenotype correlation was performed in patients. MRI was carried out within 12 h of symptoms onset and at 72 ± 12 h. The presence of HT was determined on the second DWI sequence and classified according to ECASS II criteria. MMP-9 levels were analyzed at admission. Forty-nine patients (45.8%) had HT. The -241 C/T (rs17878467) polymorphism was identified in the promoter region of the survivin gene. The prevalence of the mutant allele (T) was similar in patients and controls (14 vs. 16%, respectively; P = 0.37). However, 9 (29%) patients with allele T had HT compared to 40 (52.6%) of wild-type (P = 0.021). Logistic regression analysis showed that the polymorphism was associated with a lower risk of HT (OR 0.16; 95% CI 0.04-0.65; P = 0.01). The -241 C/T polymorphism in the promoter region of the survivin gene is associated with a lower risk of HT in patients with ischemic stroke. It has recently been reported that the -241 C/T polymorphism increases survivin promoter activity, reinforcing the hypothesis that patients with the mutant allele may have increased survivin expression in the brain. Different mechanisms, including BBB protection by the inhibition or activation of different angiogenic growth factors and the inhibition of apoptosis during

  20. Nuclear survivin and its relationship to DNA damage repair genes in non-small cell lung cancer investigated using tissue array.

    Directory of Open Access Journals (Sweden)

    Songliu Hu

    Full Text Available PURPOSE: To investigate the predictive role and association of nuclear survivin and the DNA double-strand breaks repair genes in non-small cell lung cancer (NSCLC: DNA-dependent protein kinase catalytic subunit (DNA-PKcs, Ku heterodimeric regulatory complex 70-KD subunit (Ku70 and ataxia-telangiectasia mutated (ATM. METHODS: The protein expression of nuclear survivin, DNA-PKcs, Ku70 and ATM were investigated using immunohistochemistry in tumors from 256 patients with surgically resected NSCLC. Furthermore, we analyzed the correlation between the expression of nuclear survivin, DNA-PKcs, Ku70 and ATM. Univariate and multivariate analyses were performed to determine the prognostic factors that inuenced the overall survival and disease-free survival of NSCLC. RESULTS: The expression of nuclear survivin, DNA-PKcs, Ku70 and ATM was significantly higher in tumor tissues than in normal tissues. By dichotomizing the specimens as expressing low or high levels of nuclear survivin, nuclear survivin correlated significantly with the pathologic stage (P = 0.009 and lymph node status (P = 0.004. The nuclear survivin levels were an independent prognostic factor for both the overall survival and the disease-free survival in univariate and multivariate analyses. Patients with low Ku70 and DNA-PKcs expression had a greater benefit from radiotherapy than patients with high expression of Ku70 (P = 0.012 and DNA-PKcs (P = 0.02. Nuclear survivin expression positively correlated with DNA-PKcs (P<0.001 and Ku70 expression (P<0.001. CONCLUSIONS: Nuclear survivin may be a prognostic factor for overall survival in patients with resected stage I-IIIA NSCLC. DNA-PKcs and Ku70 could predict the effect of radiotherapy in patients with NSCLC. Nuclear survivin may also stimulates DNA double-strand breaks repair by its interaction with DNA-PKcs and Ku70.

  1. 胃癌组织中Survivin、TrkB和BDNF的表达及意义%Expression and significance of survivin, TrkB and BDNF in gastric carcinoma

    Institute of Scientific and Technical Information of China (English)

    刘天卿; 任海军; 温爽; 张众

    2009-01-01

    目的 观察生存素基因蛋白(Survivin)、酪氨酸激酶受体B(TrkB)及其配体脑源性神经营养因子(BDNF)在胃癌组织和癌旁黏膜中的表达情况,探讨和分析Survivin、TrkB和BDNF与胃癌临床病理学参数的关系.方法 采用免疫组化SP法检测64例原发性胃癌组织、癌旁黏膜组织和34例淋巴结癌转移组中对应的阳性淋巴结Survivin、TrkB和BDNF蛋白的表达,分析其与临床病理学特征的关系.结果 胃癌组织中Survivin、TrkB和BDNF蛋白的阳性表达率分别为71.87%(46/64)、60.93%(39/64)和59.37%(38/64),而癌旁黏膜组织无一例表达.Survivin、TrkB和BDNF蛋白表达与患者性别、年龄、肿瘤分化程度等无关(P>0.05),而与浸润深度、淋巴结转移和TNM分期有关.浸润至胃壁全层组、有淋巴结转移组和TNM分期Ⅲ~Ⅳ组的Survivin、TrkB和BDNF阳性表达率明显高于未浸润至胃壁全层组、无淋巴结转移组和TNM分期Ⅰ~Ⅱ组(分别P0.05).结论 Survivin、TrkB和BDNF表达与胃癌发生发展密切相关,联合检测Survivin、TrkB和BDNF可有助于判断胃癌局部侵袭和远处转移的能力.

  2. Survivin-T34A: molecular mechanism and therapeutic potential

    Directory of Open Access Journals (Sweden)

    Jonathan R Aspe

    2010-12-01

    Full Text Available Jonathan R Aspe, Nathan R WallCenter for Health Disparities Research and Molecular Medicine, Division of Biochemistry and Microbiology, Department of Basic Sciences, Loma Linda University, Loma Linda, CA, USAAbstract: The inhibitor of apoptosis protein survivin's threonine 34 to alanine (T34A mutation abolishes a phosphorylation site for p34(cdc2–cyclin B1, resulting in initiation of the mitochondrial apoptotic pathway in cancer cells; however, it has little known direct effects on normal cells. The possibility that targeting survivin in this way may provide a novel approach for selective cancer gene therapy has yet to be fully evaluated. Although a flurry of work was undertaken in the late 1990s and early 2000s, only minor advances on this mutant have recently taken place. We recently described that cells generated to express a stable form of the mutant protein released this survivin-T34A to the conditioned medium. When this conditioned medium was collected and deposited on naive tumor cells, conditioned medium T34A was as effective as some chemotherapeutics in the induction of tumor cell apoptosis, and when combined with other forms of genotoxic stressors potentiated their killing effects. We hope with this review to revitalize the T34A field, as there is still much that needs to be investigated. In addition to determining the therapeutic dose and the duration of drug therapy required at the disease site, a better understanding of other key factors is also important. These include knowledge of target cell populations, cell-surface receptors, changes that occur in the target tissue at the molecular and cellular level with progression of the disease, and the mechanism and site of therapeutic action.Keywords: survivin, T34A, apoptosis, proliferation, therapy

  3. Survivin Modulates Squamous Cell Carcinoma-Derived Stem-Like Cell Proliferation, Viability and Tumor Formation in Vivo

    Directory of Open Access Journals (Sweden)

    Roberta Lotti

    2016-01-01

    Full Text Available Squamous Cell Carcinoma-derived Stem-like Cells (SCC-SC originate from alterations in keratinocyte stem cells (KSC gene expression and sustain tumor development, invasion and recurrence. Since survivin, a KSC marker, is highly expressed in SCC-SC, we evaluate its role in SCC-SC cell growth and SCC models. Survivin silencing by siRNA decreases clonal growth of SCC keratinocytes and viability of total, rapidly adhering (RAD and non-RAD (NRAD cells from primary SCC. Similarly, survivin silencing reduces the expression of stem cell markers (OCT4, NOTCH1, CD133, β1-integrin, while it increases the level of differentiation markers (K10, involucrin. Moreover, survivin silencing improves the malignant phenotype of SCC 3D-reconstruct, as demonstrated by reduced epidermal thickness, lower Ki-67 positive cell number, and decreased expression of MMP9 and psoriasin. Furthermore, survivin depletion by siRNA in RasG12V-IκBα-derived tumors leads to smaller tumor formation characterized by lower mitotic index and reduced expression of the tumor-associated marker HIF1α, VEGF and CD51. Therefore, our results indicate survivin as a key gene in regulating SCC cancer stem cell formation and cSCC development.

  4. Cooperation of bisphenol A and leptin in inhibition of caspase-3 expression and activity in OVCAR-3 ovarian cancer cells.

    Science.gov (United States)

    Ptak, Anna; Rak-Mardyła, Agnieszka; Gregoraszczuk, Ewa L

    2013-09-01

    This study was designed to investigate the effect of bisphenol A and leptin on caspase-3 expression and activity in OVCAR-3 ovarian cancer cells. Caspase-3 and survivin expression was measured at the transcript level by real-time PCR and at the protein level by Western blotting. In addition, caspase-3 activity was measured, using a fluorometric assay, upon exposure to bisphenol A (40 nM) alone, leptin (2.5 nM) alone, and the combination of both agents. 17β-estradiol (40 nM) was used as a positive control for estrogenic properties of bisphenol A. Results showed that the interaction between bisphenol A and leptin, which was similar to that observed between 17β-estradiol and leptin, led to the inhibition of caspase-3 expression and activity in OVCAR-3 cells. Surprisingly, survivin was found to not be involved in the anti-apoptotic activity of either agent. Also, results showed that leptin inhibits caspase-3 activity by acting on the signal transducers and activators of transcription 3 (STAT3) pathway, but bisphenol A and 17β-estradiol by the extracellular-signal-regulated kinases 1/2 (ERK1/2) pathway. In conclusion, the study reveals that bisphenol A and leptin interact to inhibit caspase-3 expression and activity by modulating STAT3 and ERK1/2 signaling pathways in OVCAR-3 cells.

  5. Survivin、Smad4/dpc4、APC基因在大肠腺瘤和腺癌中的表达及意义%The expression and significance of Survivin, Smad4/dpc4, APC gene in large intestinal adenoma and adenocarcinoma

    Institute of Scientific and Technical Information of China (English)

    相霞; 韦统友; 马江伟; 战则凤; 李曙晖; 郭洪飞

    2010-01-01

    目的 探讨Survivin、Smad4/dpo4、APC基因在大肠腺瘤、腺癌中的表达及意义.方法 采用免疫组化SP法检测40例正常大肠组织、80例腺瘤、80例腺癌中三种基因表达情况.结果 Survivin基因在正常大肠组织、腺瘤、腺癌中阳性表达率分别为0、35.0%、75.0%;Smad4基因为100%、95.0%、78.8%;APC为100%、80.0%、45.0%.结论 Survivin基因检测可以作为大肠癌早期诊断、观察疗效和监测肿瘤术后复发、转移等生物学行为的一个新指标.Smad4基因突变或缺失不仅诱导大肠癌的发生还促进其发展.检测APC基因突变,有助于对肿瘤病因学、发病机制及早期诊断的研究.%Objective To observe the solo-allied expression of Survivin, Smad4/dpc4 and APC gene, and study the relationship between the three genes and biological behavior of colorectal cancer. Methods The expressions of Survivin, Smad4/dpc4 and APC gene were detected by SP immunohistochem-istry among the cases from 40 eu-intestine ,80 large intestinal adenoma and 80 CRC. Results The positive rate of survivin protein in the eu-intestine, the large intestinal adenoma and CRC was 0,35.0% ,75.0% , respectively. The positive rate of Smad4/dpc4 protein was 100% , 95. 0% , 78. 8% , and the positive rate of APC protein was 100% ,80.0% ,45.0% , respectively. In the eu-intestine, APC + Smad4/dpc4 Z- expressed in 40 cases, with the incidence of 100%. In the large intestinal adenoma, APC + Smad4 expressed in 38 patients, with the incidence of 47. 5% , and the three genes expressed in 29 patients with the incidence of 23. 8%. In the CRC, two genes expressed in 57 patients, with the incidence of 71. 3% , and three genes expressed in 28 patients, with the incidence of 35.0%. Conclusion The detection of survivin was a new indicator in early diagnosis of CRC. It was significant in the diagnosis of CRC that energetic search for the positive rate level realm of survivin. The catastrophe or absence of Smad4/dpc4 gene not

  6. Survivin、b-FGF和Ang-1在大肠癌中的表达及与肿瘤微血管密度的关系%The protein expression of Survivin, b-FGF and Ang-1 in tissues of colorectal cancer and relation with tumor angiogenesis

    Institute of Scientific and Technical Information of China (English)

    赵磊; 樊微微; 高善玲

    2012-01-01

    目的 检测大肠癌组织中Survivin、b-FGF和Ang-1的表达水平,探讨Survivin和b-FGF、Ang-1与大肠癌肿瘤血管生成的关系.方法 采用免疫组织化学方法检测Survivin、b-FGF、Ang-1和CD34在7例肠炎组织、10例肠息肉组织和26例大肠癌组织中的表达.结果 Survivin和Ang-1蛋白在大肠癌组阳性表达率分别为76.9%和80.8%,高于大肠息肉组与肠炎组的表达(P<0.05).b-FGF蛋白在大肠癌组阳性表达率为73.1%,高于肠炎组(P<0.05),而与大肠息肉组比较无统计学意义(P>0.05).CD34蛋白在大肠癌组中的微血管计数(MVD)值为23.69±9.96,高于大肠息肉组与肠炎组.Survivin、b-FGF和Ang-1蛋白阳性表达肿瘤组织中多见肿瘤微血管计数增加,二者呈正相关.Survivin和促血管形成因子b-FGF、Ang-1在大肠癌组织中的表达密切相关.结论 在大肠癌发生、发展中,Survivin、b-FGF、Ang-1于肿瘤血管形成过程中起着重要的协同作用.%Objective To investigate the expression of Survivin,basic fibroblast growth factor and angiopoietin-1 in colorectal cancer and delineate the relationship of expression levels of protein with tumor angiogenesis.Methods The expression of Survivin,b-FGF,Ang-1 and CD34 were detected in 7 cases of colitis tissues,10 cases of intestinal polyps and 26 cases of colorectal cancer tissues by immunohistochemical methods.Results In colorectal cancer group,the positive expression rates of Survivin and Ang-1 protein were 76.9% and 80.8%,respectively,which were significantly higher than that in both colorectal polyps group and colitis group (P<0.05).In colorectal cancer group,the positive expression rate of b-FGF protein was 73.1%,which was significantly higher than that in colitis group (P < 0.05),but which was not statistical significance in comparison with colorectal polyps group (P > 0.05).In colorectal cancer group,CD34 protein microvessel count was 23.69±9.96,which was significantly higher

  7. Survivin和MMP-9在子宫上皮和间质细胞中的表达与子宫内膜异位症间的关系%Expression of Survivin and MMP-9 in endometrium glandular epithelium and interstitial cells and its relationship with endometriosis

    Institute of Scientific and Technical Information of China (English)

    林红霞; 张弛; 吴庆田; 马淑霞; 杨景云; 罗佳滨

    2011-01-01

    Objective To study the expression and effect of Survivin and MMP-9 in glandular epithelium and interstitial cells of eutopic and ectopic endomembrane of solenoma, abdominal ectopic and aberrant ovary, and its relationships with endometriosis. Method The expression of Survivin and MMP-9 in each sample were dectected by immunohistochemical method. Result ( 1 ) In normal endometrium, both Survivin and MMP-9 showed weak expression or no expression. In the three EMS groups, the expressions of Survivin and MMP-9 were up in varying degrees respectively, in either eutopic or ectopic endometrium, with significant differences compared to normal endometrium tissue ( P < 0.05 ). (2) In AM, AWEMS and OEMS groups, higher expression of Survivin and MMP-9 only in the proliferative phase of ectopic membrane epithelial cells with statistically significant differences compared to that in the same cells of eutopic endometrium (P <0.05); the expression in secretion phase was irregular.(3) In AM, AWEMS and OEMS groups, the expression levels of Survivin and MMP-9 in proliferative and secretory phases in tissue with same area and cell type were not significantly different ( P > 0. 05 ) and not periodic. ( 4 ) In AM, AWEMS and OEMS groups, by defining the same physical phase and tissues,the expression of Survivin and MMP-9 were much higher in glandular epithelium cells than in interstitiai cells, with statistically significant differences (P < 0.05 ). (5) In the three EMS groups, upon defining the same physical phase, tissues and cell type, the Survivin expression showed no significant differences between groups ( P > O. 05); the expression of MMP-9 in ectopic endometrial cell during secretion phase was higher in AWEMS epithelial (4.45 ±0. 18) and AM epithelial cells (4. 68 ± 0. 17) than that in the OEMS ectopic endometrial glandular epithelial cells (2. 13 ± 0. 12 ), and the differences were statistically significant (P < 0.05). Conclusion The high expression of Survivin

  8. Rapamycin-mediated mTOR inhibition attenuates survivin and sensitizes glioblastoma cells to radiation therapy

    Institute of Scientific and Technical Information of China (English)

    Arunkumar Anandharaj; Senthilkumar Cinghu; Woo-Yoon Park

    2011-01-01

    Survivin, an antiapoptotic protein, is elevated in most malignancies and attributes to radiation resistance in tumors including glioblastoma multiforme. The downregulation of survivin could sensitize glioblastoma ceils to radiation therapy. In this study, we investigated the effect of rapamycin, an inhibitor of mammalian target of rapamycin (mTOR), in attenuating survivin and enhancing the therapeutic efficacy for glioblastoma cells, and elucidated the underlying mechanisms. Here we tested various concentrations of rapamycin (1-8 nM) in combination with radiation dose 4 Gy. Rapamycin effectively modulated the protein kinase B (Akt)/mTOR pathway by inhibiting the phosphorylation of Akt and mTOR proteins, and this inhibition was further enhanced by radiation. The expression level of survivin was decreased in rapamycin pre-treatment glioblastoma ceils followed by radiation; meanwhile, the phosphorylation of H2A histone family member X (H2AX) at serine-139 (γ-H2AX) was increased, p21 protein was also induce on radiation with rapamycin pre-treatment, which enhanced G1 arrest and the accumulation of cells at G0/subG1 phase. Furthermore, the clonogenic cell survival assay revealed a significant dose-dependent decrease in the surviving fraction for all three cell lines pre-treated with rapamycin. Our studies demonstrated that targeting survivin may be an effective approach for radiosensitization of malignant glioblastoma.

  9. A Therapeutic Role for Survivin in Mitigating the Harmful Effects of Ionizing Radiation

    Directory of Open Access Journals (Sweden)

    Katherine H. Carruthers

    2016-01-01

    Full Text Available Background. Radiation therapy is a form of adjuvant care used in many oncological treatment protocols. However, nonmalignant neighboring tissues are harmed as a result of this treatment. Therefore, the goal of this study was to induce the production of survivin, an antiapoptotic protein, to determine if this protein could provide protection to noncancerous cells during radiation exposure. Methods. Using a murine model, a recombinant adenoassociated virus (rAAV was used to deliver survivin to the treatment group and yellow fluorescence protein (YFP to the control group. Both groups received targeted radiation. Visual inspection, gait analysis, and tissue histology were used to determine the extent of damage caused by the radiation. Results. The YFP group demonstrated ulceration of the irradiated area while the survivin treated mice exhibited only hair loss. Histology showed that the YFP treated mice experienced dermal thickening, as well as an increase in collagen that was not present in the survivin treated mice. Gait analysis demonstrated a difference between the two groups, with the YFP mice averaging a lower speed. Conclusions. The use of gene-modification to induce survivin expression in normal tissues allows for the protection of nontarget areas from the negative side effects normally associated with ionizing radiation.

  10. 磁性小干扰RNA纳米微粒的制备及其转染人膀胱癌细胞对沉默survivin基因表达影响的研究%Construction of magnetic small interfering RNA nanoparticles and effects of its transfection on silencing survivin gene expression and inducing human bladder cancer cell apoptosis

    Institute of Scientific and Technical Information of China (English)

    曹正国; 孙友文; 诸禹平; 苏红; 吴斌; 刘继红; 周四维

    2008-01-01

    [Objective]To investigate the construction of the magnetic smull interfering RNA(siRNA)plasmid nanoparticles and the effects of its transfection on silencing survivin gene expression and inducing bnmarx bladder cancer cells apoptosis in combination with external magnetic fields in vivo.[Methods]The siRNA-survivin recomobinant plasmid specific targeted survivin was synthesized in previous experiment.DMN were prepared by chemical coprecipitation method and used as a gene carrier.The magnetic siRNA-survivin-DMN were constructed by static electricity of polylysine and transferred into human bladder cancer BIU-87 cells with the help of external magnetic fields.The growth inhibitory rate (IR)% of BIU-87 cells Was observed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide's test and the apoptosis index(AI)% was detected by transferase-mediated dUTP nick end labeling method.The relatively transcription levels of survivin mRNA and protein expression were respectively detected by semi-quantitive reverse transcription polymerase chain reaction and Western Blotting techniques.[Resuits]The diameter,effective diameter and saturation magnetization of magnetic siRNA-DMN were about 10~12 nm,94.8 nm and 0.19 emu/S,respectively.The IR(39.60%)and AI(28.72%)in BIU-87 cells treated by siRNADMN in combination with external magnetic fields were significantly higher than those in the control group and sinsle siRNA-DMN group,respectively(P<0.05),while the relative expression of survivin mRNA and protein was significantly lower(P<0.05)[Conclusion]The magnetic siRNA-DMN plasmid nanopartieles could effectively inhibit survivin expression and induce BIU-87 cells apoptosis after transferred into cells in combination with extemal magnetic fields which provided experimental basis for the magnetic targeting gene therapy of bladder tumor.%目的 研究磁性小干扰RNA(siRNA)重组质粒纳米微粒的制备及其在外磁场协同作用下转染人膀胱癌细胞对膀胱癌

  11. 碘染色联合p53、Survivin及端粒酶检测对早期食管癌及癌前病变的诊断价值%The role of endoscopy iodine staining combined detecting the expression of p53 ,survivin and telomerase in the diagnosis of early e-sophageal carcinoma and precancerom lesions

    Institute of Scientific and Technical Information of China (English)

    刘春安; 张新东; 杨艳; 马凯; 白景芝; 王祥阁; 王静; 柴同海

    2009-01-01

    Objective To study the value of using endoscopy iodine staining combined detecting the expression of p53, survivin and telomerase in the diagnosis of early esophageal carcinoma and precancerous lesions. Methods Three hundred patients who received routine gastrescopy and had high risk of esophageal carcinoma were examined by the iodine dyeing on their esophageal mucosa with 2. 5% Lugol solu-tion. Multiple biopsies were taken from the unstained or light-stained areas when their diameters were over 5mm for pathology diagnosis. The expression of p53, survivin and telomerase were detected by immunohistochemical staining. Results With the Lugol staining method, the incidences of light, moderate and high grade dysplasia and early esophageal carcinoma were 16. 3% ,9. 7% ,3.7% and 4. 3% respectively. The expression of p53, survivin and telomerase were related and the unstained were related to the expression of p53, survivin and telomerase in dysplasia and early esophageal carcinoma. Using endoscopy iodine staining combined testing p53, survivin and telomerase showed sensitiv-ity of 100%, specificity of 92. 3% and accuracy of 91.8% to diagnose of early esophageal carcinoma and high grade dysplasia. Conclusion Lugol solution chromoendscopy combined assay of p53, survivin and telomersse might be useful in the diagnosing early esophageal carcino-ma and precancerous lesion.%目的 探讨内镜下碘染色联合组织p53、Survivin及端粒酶检测对早期食管癌及癌前病变的诊断价值.方法 本院因上消化道症状接受胃镜检查且食管黏膜有可疑病变的门诊及住院患者300例进行卢戈碘液染色.对直径≥5 mm的不染区或淡柒区取活检送病理.用免疫组化方法 对病理组织学检查证实为黏膜不典型增生、早期食管癌及30例正常对照者分别检测p53、Sur-vivin及端粒酶表达情况.结果 碘染色对食管黏膜不典型增生轻、中、重度及早期鳞状细胞癌检出率分别为16.3%、9.7%、3.7%

  12. survivin在皮肤血管瘤组织中的表达及其与半胱氨酸天冬氨酸蛋白酶-3的关系%Expression of survivin in human dermal hemangioma and its relationship with caspase-3

    Institute of Scientific and Technical Information of China (English)

    刘思洋; 叶玲; 陕声国; 张端莲

    2008-01-01

    Objective To investigate the expression of survivin and its relationship with that of caspase-3 in two stages of hemangioma and normal skin, and to explore the role of survivin in the patho-genesis of hemangioma. Methods A total of 50 cases of human dermal hemangioma and 8 eases of normal skin were analyzed for expression of survivin and caspase-3 by immunohistochemistry. Results The posi-tive ratio and average light density of survivin were obviously higher in proliferative hemangioma (0.2449±0.0135,0.7246±0.0747) than that in involutional hemangioma (0.1648±0.0217,0.5592±0.1601) and normal skin (0.1789±0.0126,0.4626±0.0961) (P<0.01). On the contrary, the expression of easpase-3 was up-regulated in involutional hemangioma (0.2386±0.0175, 0.4378±0.0593). Analysis of data also showed that there was a negative correlation between the average light density of survivin and caspase-3 (r=-0.95,P<0.01). Conclusions Survivin plays an important role in the pathogenesis of hemangioma and has a negative relationship with easpase-3.%目的 检测凋亡抑制因子survivin在各期皮肤真性血管瘤中的表达及其与半胱氨酸天冬氨酸蛋白酶-3(caspase-3)表达的关系,探讨其在血管瘤发病机制中的作用以及在临床分子治疗中的意义.方法 利用免疫组织化学SP染色法显示survivin、caspase-3在增生期、退化期血管瘤组织和正常皮肤组织中的表达,并运用病理图文报告管理系统处理染色结果,比较各组表达有无差异,分析两指标间的关系.结果 survivin在增生期血管瘤、退化期血管瘤和正常皮肤组织中的吸光度值分别是0.2449±0.0135、0.1648±0.0217和0.1789±0.0126,增生期组与其他两组间差异有统计学意义(P<0.01);而caspase-3在退化期血管瘤中的表达明显高于增生期血管瘤组织和正常皮肤(P<0.01).在增生期和退化期血管瘤组织中,survivin与caspase-3的表达呈统计学负相关性(r=-0.95、P<0.01).结论 sur-vivin通

  13. Clinical significance of the expression of serum level of CA125 and survivin protein in endometrial carcinoma%血清CA125水平及Survivin蛋白在子宫内膜癌中的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    徐行丽; 井甜甜; 贾庆兰

    2010-01-01

    目的 研究血清CA125水平及Survivin蛋白在子宫内膜癌中的表达及临床意义.方法 正常子宫内膜组(40例)、子宫内膜不典型增生组(40例)及子宫内膜癌组(40例).比较各组手术前后及子宫内膜癌组不同临床、病理分期中血清CA125水平变化及Survivin蛋白的表达情况.结果 子宫内膜癌组血清CA125水平在手术前后均明显高于其余两组,差异均有统计学意义(P<0.05).子宫内膜不典型增生组及子宫内膜癌组的Survivin阳性表达率及强阳性表达率与正常子宫内膜组,差异有统计学意义(P<0.05).血清CA125在子宫内膜癌Ⅱ-Ⅳ期、病理级别高、深肌层浸润及复发组中的阳性表达率明显升高.Survivin蛋白的强阳性表达率在Ⅱ-Ⅳ期、G3、深肌层浸润及复发病例中明显升高.结论 血清CA125水平对子宫内膜癌的手术病理分期有一定的指导意义.Survivin蛋白对子宫内膜癌的早期诊断及临床预后有一定的参考价值.%Objective To study clinical significance of the expression of serum level of CA125 and survivin protein in endometrial carcinoma. Methods The normal endometrium group(40 cases) ,atypical endometrial hyperplasia group (40 cases)and endometrial cancer group (40 cases). Compared the changes of serum level of CA125 and the expression of survivin protein between before and after surgery in each group and in different clinical and pathological stagings in the group of endometrial carcinoma. Results The serum level of CA125in endometrial cancer group before and after surgery are significantly higher than other two groups, the differences are statistically significant(P < 0.05 ). Compare survivin positive expression rate and strong positive expression rate in atypical endometrial hyperplasia group and endometrial cancer group with the normal endometrium group, the differences are statistically significant( P < 0.05 ). The rate of positive expression of serum level of CA125 in

  14. survivin特异性RNAi表达载体的构建及其干扰效果鉴定%Construction and effect identification of RNAi eukaryotic expression vectors of survivin

    Institute of Scientific and Technical Information of China (English)

    汪欣; 杨军; 许波; 熊宇; 张从纪

    2009-01-01

    目的 构建survivin基因的RNAi真核表达载体并观察其对转染腺样囊性癌细胞株ACC-2后survivin表达变化以及对细胞凋亡的影响.方法 设计、合成siRNA(small interfering RNA),构建表达载体pGenesil-shRNA-survivin,将重组质粒导入人腺样囊性癌ACC-2细胞株.采用逆转录-聚合酶链反应(RT-PCR)法检测转染后的ACC-2细胞中survivin mRNA表达的变化,用TUNEL检测细胞凋亡的变化.结果 经测序模版序列和设计序列完全正确,在pGenesil-shRNA-survivin转载ACC-2细胞株中,survivin mRNA表达明显降低,转染的ACC-2细胞凋亡显著增加.结论 成功构建了survivin基因的RNAi真核表达载体,且其对腺样囊性癌细胞有一定的干扰效果.

  15. Identification of novel helper epitope peptides of Survivin cancer-associated antigen applicable to developing helper/killer-hybrid epitope long peptide cancer vaccine.

    Science.gov (United States)

    Ohtake, Junya; Ohkuri, Takayuki; Togashi, Yuji; Kitamura, Hidemitsu; Okuno, Kiyotaka; Nishimura, Takashi

    2014-09-01

    We identified novel helper epitope peptides of Survivin cancer antigen, which are presented to both HLA-DRB1*01:01 and DQB1*06:01. The helper epitope also contained three distinct Survivin-killer epitopes presented to HLA-A*02:01 and A*24:02. This 19 amino-acids epitope peptide (SU18) induced weak responses of Survivin-specific CD4(+) and CD8(+) T cells though it contained both helper and killer epitopes. To enhance the vaccine efficacy, we synthesized a long peptide by conjugating SU18 peptide and another DR53-restricted helper epitope peptide (SU22; 12 amino-acids) using glycine-linker. We designated this artificial 40 amino-acids long peptide containing two helper and three killer epitopes as Survivin-helper/killer-hybrid epitope long peptide (Survivin-H/K-HELP). Survivin-H/K-HELP allowed superior activation of IFN-γ-producing CD4(+) Th1 cells and CD8(+) Tc1 cells compared with the mixture of its component peptides (SU18 and SU22) in the presence of OK-432-treated monocyte-derived DC (Mo-DC). Survivin-H/K-HELP-pulsed Mo-DC pretreated with OK-432 also exhibited sustained antigen-presentation capability of stimulating Survivin-specific Th1 cells compared with Mo-DC pulsed with a mixture of SU18 and SU22 short peptides. Moreover, we demonstrated that Survivin-H/K-HELP induced a complete response in a breast cancer patient with the induction of cellular and humoral immune responses. Thus, we believe that an artificially synthesized Survivin-H/K-HELP will become an innovative cancer vaccine.

  16. siRNA AGAINST SURVIVIN SUPPRESSES THE PROLIFERATION OF PANCREATIC CANCER CELL PC-2 AND INDUCES ITS APOPTOSIS

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Survivinis a member of the inhibitor of apop-tosis family,whichinhibits cell apoptosis and regu-lates cell division.Survivin is expressed in embry-onic tissues as well as in the majority of humancancers,but is not expressed in most nor mal adulttissues.The highly cancer-specific expression ofSurvivin,coupled withits i mportance in inhibitingcell apoptosis and in regulating cell division,makesit a useful diagnostic marker and ideal target forcancer treat ment[1].RNAi is a process of post transcriptional gene...

  17. Expressions of Survivin, COX-2 in GPL and gastric carcinoma and their association with helicobactor pyiori infection%胃癌及前病变中 Survivin 和 COX-2 蛋白的表达及其与Hp感染的关系

    Institute of Scientific and Technical Information of China (English)

    姬琛华; 刘丽娜; 吕申; 孟华

    2008-01-01

    [目的]分析胃癌及癌前病变survivin、COX-2蛋白表达与幽门螺杆菌(Helicobacter pylori,H pylori)感染的关系,探讨胃癌发病的可能机制.[方法]收集胃癌标本51例,非癌胃黏膜标本47例,应用快速尿素酶试验与W-S银染色法将各种病变分为Hp感染阳性组和阴性组,应用免疫组化SP法时各种病变survivin、COX-2蛋白的表达进行检测.采用X2检验进行数据分析.[结果]Survivin蛋白的表达从慢性浅表性胃炎(CSG)到胃癌(GC)逐渐增加,其中慢性萎缩性胃炎(CAG)组明显低于肠化生(IM)、不典型增生(Dys)和胃癌(GC)组(Survivin 30.5%VS60.0%、62.5%、70.6%,P<0.05),而IM、Oys组与GC组差异无显著性意义(P0.05);COX-2蛋白在CSG、CAG、IM、Dys、GC中的表达率分别为0%、23.1%、60.0%、50.0%、68.6%.其中,CAG组亦明显低于IM、Dys、GC组(P<0.05),而IM、Dys组与GC组差异无显著性意义(P0.05).在GC组,Hp阳性者Survivin、COX-2表达明显高于Hp阴性者(Survivin 70.6%VS 29.4%,COX-2 68.6%VS 31.4%,P<0.01).[结论]Survivin、COX-2可能参与胃癌的早期形成,Hp感染可能是其过度表达的机制之一.

  18. The inhibition of the apoptosis pathway by the Coxiella burnetii effector protein CaeA requires the EK repetition motif, but is independent of survivin.

    Science.gov (United States)

    Bisle, Stephanie; Klingenbeck, Leonie; Borges, Vítor; Sobotta, Katharina; Schulze-Luehrmann, Jan; Menge, Christian; Heydel, Carsten; Gomes, João Paulo; Lührmann, Anja

    2016-05-18

    ABSRTACT Coxiella burnetii is an obligate intracellular bacterium that causes Query (Q) fever, a zoonotic disease. It requires a functional type IV secretion system (T4SS) which translocate bacterial effector proteins into the host cell cytoplasm and thereby facilitates bacterial replication. To date, more than 130 effector proteins have been identified, but their functions remain largely unknown. Recently, we demonstrated that one of these proteins, CaeA (CBU1524) localized to the host cell nucleus and inhibited intrinsic apoptosis of HEK293 or CHO cells. In the present study we addressed the question whether CaeA also affects the extrinsic apoptosis pathway. Ectopic expression of CaeA reduced extrinsic apoptosis and prevented the cleavage of the executioner caspase 7, but did not impair the activation of initiator caspase 9. CaeA expression resulted in an up-regulation of survivin (an inhibitor of activated caspases), which, however, was not causal for the anti-apoptotic effect of CaeA. Comparing the sequence of CaeA from 25 different C. burnetii isolates we identified an EK (glutamic acid/ lysine) repetition motif as a site of high genetic variability. The EK motif of CaeA was essential for the anti-apoptotic activity of CaeA. From these data, we conclude that the C. burnetii effector protein CaeA interferes with the intrinsic and extrinsic apoptosis pathway. The process requires the EK repetition motif of CaeA, but is independent of the upregulated expression of survivin.

  19. Protection effect of survivin protein overexpression on acute myocardial infarction in rats.

    Science.gov (United States)

    Yang, Meng; Li, Bo; Liu, Jingwei; Sun, Haiyan

    2015-01-01

    To investigate the protective effect of adenovirus mediated Survivin protein overexpression on acute myocardial infarction in rats. 45 acute myocardial infarction rat models were constructed by suture method and were randomly divided into sham group, model group and treatment group. The treatment group was injected with Survivin gene packed virus via ventricle. The model group was injected with equal titer of adenovirus packed empty vector. The sham group was not ligated. These rats were killed in 96 h after treatment. The levels of Survivin, Caspase-3, caspase-7 mRNA and protein in myocardial tissues were detected by real-time fluorescence quantitative PCR and Western blot. Myocardium tissue cell apoptosis were analyzed by TUNEL staining, the immunology of myocardial infarction tissue was analyzed by TTC staining. Compared with model group and sham group, the level of survivin protein in myocardium tissue of rats in treatment group was significantly increased (Pmyocardial tissue of rats in model group and treatment group were significantly increased, but the treatment group were significantly lower than those of model group (Pmyocardial infarction areas of rats in model group and treatment group were significantly higher than those of sham group, but the treatment group were significantly lower than those of model group (Pmyocardial tissue can significantly inhibit the expression of apoptosis promoting factor in myocardial tissue of acute myocardial infarction rats, reduce the apoptosis index of myocardial cells and the myocardial infarct size, which has great significance for protecting myocardial function.

  20. Regulation of Survivin and CDK4 by Epstein-Barr virus encoded latent membrane protein 1 in nasopharyngeal carcinoma cell lines

    Institute of Scientific and Technical Information of China (English)

    Mi Dan AI; Li Li LI; Xiao Rong ZHAO; Yong WU; Jian Ping GONG; Ya CAO

    2005-01-01

    Latent membrane protein 1 (LMP1), an important protein encoded by Epstein Barr virus (EBV), has been implied to link with the pathogenesis of nasopharyngeal carcinoma (NPC). Its dual effects of increasing cell proliferation and inhibiting cell apoptosis have been confirmed. In this study, we showed that the expression of Survivin and CDK4 protein in CNE-LMP1, a LMP1 positive NPC epithelial cell line, is higher than in LMP1 negative NPC epithelial cell line CNE1, and the expression is LMP1 dosage-dependent. Although it was reported that Survivin specifically expressed in cell cycle G2/M phase, our studies suggested that LMP1 could promote the expression of Survivin in G0/G1, S and G2/M phase. It also showed that Survivin and CDK4 could be accumulated more in the nuclei triggered by LMP1. More interestingly, Survivin and CDK4 could form a protein complex in the nuclei of CNE-LMP1 rather than in that of CNE1, which demonstrated that the interaction between these two proteins could be promoted by LMP1. These results strongly suggested that the role of LMP1 in the regulation of Survivin and CDK4 may also shed some light on the mechanism research of LMP1 in NPC.

  1. Validation of cytoplasmic-to-nuclear ratio of survivin as an indicator of improved prognosis in breast cancer

    LENUS (Irish Health Repository)

    Rexhepaj, Elton

    2010-11-23

    Abstract Background Conflicting data exist regarding the prognostic and predictive impact of survivin (BIRC5) in breast cancer. We previously reported survivin cytoplasmic-to-nuclear ratio (CNR) as an independent prognostic indicator in breast cancer. Here, we validate survivin CNR in a separate and extended cohort. Furthermore, we present new data suggesting that a low CNR may predict outcome in tamoxifen-treated patients. Methods Survin expression was assessed using immunhistochemistry on a breast cancer tissue microarray (TMA) containing 512 tumours. Whole slide digital images were captured using an Aperio XT scanner. Automated image analysis was used to identify tumour from stroma and then to quantify tumour-specific nuclear and cytoplasmic survivin. A decision tree model selected using a 10-fold cross-validation approach was used to identify prognostic subgroups based on nuclear and cytoplasmic survivin expression. Results Following optimisation of the staining procedure, it was possible to evaluate survivin protein expression in 70.1% (n = 359) of the 512 tumours represented on the TMA. Decision tree analysis predicted that nuclear, as opposed to cytoplasmic, survivin was the most important determinant of overall survival (OS) and breast cancer-specific survival (BCSS). The decision tree model confirmed CNR of 5 as the optimum threshold for survival analysis. Univariate analysis demonstrated an association between a high CNR (>5) and a prolonged BCSS (HR 0.49, 95% CI 0.29-0.81, p = 0.006). Multivariate analysis revealed a high CNR (>5) was an independent predictor of BCSS (HR 0.47, 95% CI 0.27-0.82, p = 0.008). An increased CNR was associated with ER positive (p = 0.045), low grade (p = 0.007), Ki-67 (p = 0.001) and Her2 (p = 0.026) negative tumours. Finally, a high CNR was an independent predictor of OS in tamoxifen-treated ER-positive patients (HR 0.44, 95% CI 0.23-0.87, p = 0.018). Conclusion Using the same threshold as our previous study, we have

  2. Validation of cytoplasmic-to-nuclear ratio of survivin as an indicator of improved prognosis in breast cancer

    Directory of Open Access Journals (Sweden)

    Duffy Michael J

    2010-11-01

    Full Text Available Abstract Background Conflicting data exist regarding the prognostic and predictive impact of survivin (BIRC5 in breast cancer. We previously reported survivin cytoplasmic-to-nuclear ratio (CNR as an independent prognostic indicator in breast cancer. Here, we validate survivin CNR in a separate and extended cohort. Furthermore, we present new data suggesting that a low CNR may predict outcome in tamoxifen-treated patients. Methods Survin expression was assessed using immunhistochemistry on a breast cancer tissue microarray (TMA containing 512 tumours. Whole slide digital images were captured using an Aperio XT scanner. Automated image analysis was used to identify tumour from stroma and then to quantify tumour-specific nuclear and cytoplasmic survivin. A decision tree model selected using a 10-fold cross-validation approach was used to identify prognostic subgroups based on nuclear and cytoplasmic survivin expression. Results Following optimisation of the staining procedure, it was possible to evaluate survivin protein expression in 70.1% (n = 359 of the 512 tumours represented on the TMA. Decision tree analysis predicted that nuclear, as opposed to cytoplasmic, survivin was the most important determinant of overall survival (OS and breast cancer-specific survival (BCSS. The decision tree model confirmed CNR of 5 as the optimum threshold for survival analysis. Univariate analysis demonstrated an association between a high CNR (>5 and a prolonged BCSS (HR 0.49, 95% CI 0.29-0.81, p = 0.006. Multivariate analysis revealed a high CNR (>5 was an independent predictor of BCSS (HR 0.47, 95% CI 0.27-0.82, p = 0.008. An increased CNR was associated with ER positive (p = 0.045, low grade (p = 0.007, Ki-67 (p = 0.001 and Her2 (p = 0.026 negative tumours. Finally, a high CNR was an independent predictor of OS in tamoxifen-treated ER-positive patients (HR 0.44, 95% CI 0.23-0.87, p = 0.018. Conclusion Using the same threshold as our previous study, we have

  3. Inhibition of signal transducer and activator of transcription 3 expression by RNA interference suppresses invasion through inducing anoikis in human colon cancer cells

    Institute of Scientific and Technical Information of China (English)

    Yu Fan; You-Li Zhang; Ying Wu; Wei Zhang; Yin-Huan Wang; Zhao-Ming Cheng; Hua Li

    2008-01-01

    AIM: To investigate the roles and mechanism of signal transducer and activator of transcription 3 (STAT3) in invasion of human colon cancer cells by RNA interference. METHODS: Small interfering RNA (siRNA) targeting Signal transducer and activator of transcription 3 (STAT3) was transfected into HT29 colon cancer cells. STAT3 protein level and DNA-binding activity of STAT3 was evaluated by western blotting and electrophoretic mobility shift assay (EMSA), respectively. We studied the anchorage-independent growth using colony formation in soft agar, and invasion using the boyden chamber model, anoikis using DNA fragmentation assay and terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL), respectively. Western blot assay was used to observe the protein expression of Bcl-xL and survivin in colon cancer HT29 cells. RESULTS: RNA interference (RNAi) mediated by siRNA leads to suppression of STAT3 expression in colon cancer cell lines. Suppression of STAT3 expression by siRNA could inhibit anchorage-independent growth, and invasion ability, and induces anoikis in the colon cancer cell line HT29. It has been shown that knockdown of STAT3 expression by siRNA results in a reduction in expression of Bcl-xL and survivin in HT29 cells. CONCLUSION: These results suggest that STAT3 siRNA can inhibit the invasion ability of colon cancer cells through inducing anoikis, which antiapoptotic genes survivin and Bcl-xL contribute to regulation of anoikis. These studies indicate STAT3 siRNA could be a useful therapeutic tool for the treatment of colon cancer.

  4. Survivin selective inhibitor YM155 induce apoptosis in SK-NEP-1 Wilms tumor cells

    Directory of Open Access Journals (Sweden)

    Tao Yan-Fang

    2012-12-01

    Full Text Available Abstract Background Survivin, a member of the family of inhibitor of apoptosis proteins, functions as a key regulator of mitosis and programmed cell death. YM155, a novel molecular targeted agent, suppresses survivin, which is overexpressed in many tumor types. The aim of this study was to determine the antitumor activity of YM155 in SK-NEP-1 cells. Methods SK-NEP-1 cell growth in vitro and in vivo was assessed by MTT and nude mice experiments. Annexin V/propidium iodide staining followed by flow cytometric analysis was used to detect apoptosis in cell culture. Then gene expression profile of tumor cells treated with YM155 was analyzed with real-time PCR arrays. We then analyzed the expression data with MEV (Multi Experiment View cluster software. Datasets representing genes with altered expression profile derived from cluster analyses were imported into the Ingenuity Pathway Analysis tool. Results YM155 treatment resulted in inhibition of cell proliferation of SK-NEP-1cells in a dose-dependent manner. Annexin V assay, cell cycle, and activation of caspase-3 demonstrates that YM155 induced apoptosis in SK-NEP-1 cells. YM155 significantly inhibited growth of SK-NEP-1 xenografts (YM155 5 mg/kg: 1.45 ± 0.77 cm3; YM155 10 mg/kg: 0.95 ± 0.55 cm3 compared to DMSO group (DMSO: 3.70 ± 2.4 cm3 or PBS group cells (PBS: 3.78 ± 2.20 cm3, ANOVA P Conclusions The present study demonstrates that YM155 treatment resulted in apoptosis and inhibition of cell proliferation of SK-NEP-1cells. YM155 had significant role and little side effect in the treatment of SK-NEP-1 xenograft tumors. Real-time PCR array analysis firstly showed expression profile of genes dyes-regulated after YM155 treatment. IPA analysis also represents new molecule mechanism of YM155 treatment, such as NR3C1 and dexamethasone may be new target of YM155. And our results may provide new clues of molecular mechanism of apoptosis induced by YM155.

  5. 3-bromopyruvate and sodium citrate target glycolysis, suppress survivin, and induce mitochondrial-mediated apoptosis in gastric cancer cells and inhibit gastric orthotopic transplantation tumor growth.

    Science.gov (United States)

    Wang, Ting-An; Zhang, Xiao-Dong; Guo, Xing-Yu; Xian, Shu-Lin; Lu, Yun-Fei

    2016-03-01

    Glycolysis is the primary method utilized by cancer cells to produce the energy (adenosine triphosphate, ATP) required for cell proliferation. Therefore, inhibition of glycolysis may inhibit tumor growth. We previously found that both 3-bromopyruvate (3-BrPA) and sodium citrate (SCT) can inhibit glycolysis in vitro; however, the underlying inhibitory mechanisms remain unclear. In the present study, we used a human gastric cancer cell line (SGC-7901) and an orthotopic transplantation tumor model in nude mice to explore the specific mechanisms of 3-BrPA and SCT. We found that both 3-BrPA and SCT effectively suppressed cancer cell proliferation, arrested the cell cycle, induced apoptosis, and decreased the production of lactate and ATP. 3-BrPA significantly reduced the glycolytic enzyme hexokinase activity, while SCT selectively inhibited phosphofructokinase-1 activity. Furthermore, 3-BrPA and SCT upregulated the expression of pro-apoptotic proteins (Bax, cytochrome c, and cleaved caspase-3) and downregulated the expression of anti-apoptotic proteins (Bcl-2 and survivin). Finally, our animal model of gastric cancer indicated that intraperitoneal injection of 3-BrPA and SCT suppressed orthotopic transplantation tumor growth and induced tumor apoptosis. Taken together, these results suggest that 3-BrPA and SCT selectively suppress glycolytic enzymes, decrease ATP production, induce mitochondrial-mediated apoptosis, downregulate survivin, and inhibit tumor growth. Moreover, an intraperitoneal injection is an effective form of administration of 3-BrPA and SCT.

  6. The effect of Survivin antioligonucleotide on the apoptosis of Xuanwei lung adenocarcinoma cell%Survivin反义寡核苷酸对宣威肺腺癌细胞凋亡的影响

    Institute of Scientific and Technical Information of China (English)

    Weiwei Wang; Gaofeng Li; Zhipeng Hong; Ruibing Cheng; Jipeng Zhang; Lianhua Ye

    2011-01-01

    Objective:The aim of the study was to investigate the effects of Survivin antioligonucleotide on the proliferation,apoptosis and cell cycle of Xuanwei lung adenocarcinoma cell XWLC-05. Methods:Specific targeting Survivin ASODN was composed firstly. XWLC-05 would be divided into 4 groups:Group Sham (blank), Group Lip (simple liposome), Group LipSODN (transfected sense oligonucleotide) and Group Lip-ASODN (transfected ASODN). All groups had been transfected under the same condition for 48 hours. Then West blotting was used to check the expression of Survivin in cells from different groups and flow cytometer was used to find out the apoptosis rate of cells in different groups. Results:The expression of XWLC-05 Survivin in Group Lip-ASODN decreased obviously and apoptosis rate was significantly higher than that of other groups (P < 0. 05). Conclusion:XWLC-05 transfected by Survivin ASODN could down regulate the expression of Survivin and lower expression of Survivin might lead to the apoptosis of XWLC-05 and restrain the proliferation of cancer cells.

  7. 胞二磷胆碱结合康复训练对脑缺血大鼠半暗带区存活素表达的影响%The effect of drugs in combination with rehabilitative training on the expression of survivin in the ischemic penumbra after focal cerebral ischemia in rats

    Institute of Scientific and Technical Information of China (English)

    毕然然; 孙强三; 孙丽

    2013-01-01

    Objective To investigate the effect of drugs in combination with rehabilitative training on motor function and the expression of survivin in the ischemic penumbra after focal cerebral ischemia.Methods One hundred and twenty male,adult,Sprague-Dawley rats were subjected to left middle cerebral artery occlusion (MCAO)by suturing.Ninety-six of them were then randomly divided into a control group,a drug group,a rehabilitative trainiug group,and a drugs in combination with rehabilitative training group,with 24 in each.For three days the rats in the control group received no treatment,while those in the drug group received 500 mg/kg of citicoline daily,those in the rehabilitative training group received motor training including balancing,grasping,rotating and walking exercises,aud those in the drug and rehabilitative training group received both citicoline and the motor training.Behavioral tests were administered to all groups,and immunohistochemistry was used to detect the expression of survivin in the ischemic penumbra.Results Average behavior scores in the drug group and the control group were not significantly different at day 7,14 or 21 after the MCAO.Average behavior scores in the rehabilitative training and drug in combination with rehabilitative training groups were significantly superior to those of the control and drug groups at day 14 and 21.At those time points the average scores in the drug in combination with rehabilitative training group were also significantly better than those of the rehabilitation training group.Compared with control group,at the 7th,14th and 21st day after MCAO,expression of survivin in the other three groups had increased significantly.Expression of survivin in the group where drug treatment was combined with rehabilitative training was significantly greater than in the drug and rehabilitative training groups.Conclusions Citicoline in combination with rehabilitative training can improve the recovery of motor function in rats

  8. Correlation of survivin, p53 and Ki-67 in laryngeal cancer Hep-2 cell proliferation and invasion

    Institute of Scientific and Technical Information of China (English)

    Shi-Geng Pei; Ju-Xiang Wang; Xue-Ling Wang; Qing-Jun Zhang; Hong Zhang

    2015-01-01

    Objective:To investigate the mechanism of survivin, p53 and Ki-67 on Hep-2 human laryngeal cancer endothelial cell proliferation and invasion.Methods:Laryngeal squamous cell carcinoma and paracancerous normal tissues were collected, total RNA was extracted from tissues,survivin,p53and Ki-67gene mRNA expression levels in laryngeal cancer and the adjacent tissues were detected by Real-time PCR. Human laryngeal cancer Hep-2 epithelial cells were selected,survivin gene was overexpressed, and cell proliferation was detected by MTT.p53 andKi-67gene expression changes in overexpressedsurvivin gene were detected by Western blot. Changes in Hep-2 cell invasive ability were studied whensurvivin was overexpressed as detected by Transwell invasion assay.Results: In the adjacent tissues, survivin,p53andKi-67 gene relative expression levels were 1.72 ± 0.9, 13.7 ± 5.7 and 5.7 ± 1.3, respectively; while in cancer tissues, gene relative expression levels were 53.7 ± 8.3, 66.7 ± 5.2 and 61.0 ± 3.1, respectively, which was significantly increased. As detected by MTT, relative cell survival rate within 12 h ofsurvivinoverexpression were: load control group, (88.5±1.6)%; overexpressed group, (90.3±1.9)%. Transwell invasion assay results indicated that overexpressedsurvivincould significantly increase the relative survival rate of cells. Conclusions:Expressions ofp53,Ki67 and survivin are increased in cancer; and there is a positive correlation betweensurvivin, p53andKi67 expressions in laryngeal carcinoma.

  9. [Inhibition Function of Dominant-negative Mutant Gene Survivin-D53A to SPC-A1 Lung Adenocarcinoma Xenograft in Nude Mice Models].

    Science.gov (United States)

    Yu, Min; Peng, Xingchen; Lu, You; Huang, Meijuan

    2015-06-01

    Survivin-D53A (SVV-D53A) is a dominant-negative mutant survivin, which represents a potential promising target for cancer gene therapy. The present study was designed to determine whether SVV-D53A plasmid encapsuled by DOTAP: Chol liposome would have the anti-tumor activity against SPC-A1 lung adenocarcinoma, and to detect the possible mechanisms. In our experiment, SPC-A1 cells were transfected in vitro with SVV-D53A plasmid and examined for protein expression by Western blot, then flow cytometric analysis was used to detect apoptosis. SPC-A1 lung adenocarcinoma xenografts were established in vivo in the nude mice, which received the i. v. administrations of SVV-D53A plasmid/liposome complexes. After mice were sacrificed, the paraffin-embedded tumor tissue sections were used for proliferating cell nuclear antigen (PCNA) expression and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Compared with the control group, the mice treated with SVV-D53A plasmid had an obviously reduced tumor volume, with high level of apoptosis and decreased cell proliferation in tumor tissue. The research results proved that the administration of SVV-D53A plasmid resulted in significant inhibition of SPC-A1 cells both in vitro and in vivo. The functional mechanism is that the anti-tumor response causes and induces tumor cell apoptosis.

  10. Construction of miRNA expression vector dual-targeting on HIF-1α/ survivin genes and its effects on proliferation of pancreatic cancer Panc-1 cell line%抗HIF-1α和survivin基因双靶位miR-RNAi载体的构建及其对胰腺癌细胞的作用

    Institute of Scientific and Technical Information of China (English)

    徐孙兵; 朱一平; 牟一平; 朱玲华

    2012-01-01

    目的:构建抗HIF-1α单靶位(anti-H)、抗Survivin单靶位(anti-S)以及抗HIF-1α和Survivin基因双靶位微小RNA表达载体(anti-H+S),观察和比较这3种载体对胰腺癌Panc-1细胞增殖的影响.方法:设计4组靶向HIF-1α和Survivin基因的寡核苷酸序列,连接至pcDNA6.2-GW/EmGFP-miR质粒,构建两组共8个载体,依次为anti-H(Ⅰ)、anti-H(Ⅱ)、anti-H(Ⅲ)、anti-H(Ⅳ)和anti-S(Ⅰ)、anti-S(Ⅱ)、anti-S(Ⅲ)、anti-S(Ⅳ).实时定量RT-PCR检测靶基因mRNA的表达水平,将两组中mRNA下调作用最强的质粒串联,构建anti-H+S.用anti-H+S及mRNA下调作用最强的anti-H和anti-S转染胰腺癌Panc-1细胞,Western blot测定靶蛋白表达情况,MTT法检测细胞增殖活性.结果:经测序鉴定,所有anti-H、anti-S及双靶位质粒anti-H+S装载位点核苷酸序列与设计序列完全相符.anti-H的靶基因mRNA沉默效率依次为48%、2%、44%和30%;anti-S的靶基因mRNA沉默效率依次为72%、75%、58%和59%.由anti-H(Ⅰ)和anti-S(Ⅱ)串联得到双靶位质粒anti-H+S.anti-H+S的靶基因mRNA沉默效率为53% (HIF-1α)和42%(survivin).anti-H(Ⅰ)、anti-S(Ⅱ)及anti-H+S与对照质粒相比均使靶基因蛋白的表达明显下降,细胞增殖受到明显抑制(P<0.05).其中anti-H+S对细胞的增殖抑制作用强于anti-H(Ⅰ)和anti-S(Ⅱ),72h后差异有统计学意义(P<0.05).结论:本研究成功构建了抗HIF-1α和Survivin的单靶位质粒和双靶位质粒;其中,anti-H+S抑制胰腺癌Panc-1细胞增殖作用强于anti-H(Ⅰ)和anti-S(Ⅱ).%Objective:To design and construct miRNA expression vector dual-targeting on HIF-1αand survivin genes and to investigate its effects on proliferation of human pancreatic cancer cells. Methods; The specific pre-miRNA single strand DNA oligos for HIF-1α and survivin genes were designed and synthesized,then via annealing and ligating with pcDNA6. 2-GW/EmGFP-miR plasmids in order,two kinds (eight in total) of mi

  11. Apoptosis of drug-resistant human ovarian carcinoma cell line COC1/DDP induced by survivin antisense oligonucleotides

    Institute of Scientific and Technical Information of China (English)

    ZHENG Fei; RUAN Fei; XIE Xian-kuan; LIU Shao-yang

    2006-01-01

    @@ Currently, surgery-oriented treatment plays a major role in the treatment of ovarian cancer patients. But 5-year survival rate of patients is still around 30%. One of the main reasons for the Iow survival rate is the drug resistance of tumor cells against chemotherapy.1,2 The function of antiapoptosis in the course of initiation and progress of cancer has a close relationship with drug resistance of tumor cells. Survivin is a new discovered anti-apoptosis gene, its expression levels correlating with more aggressive disease and poor clinical outcome in many of these tumors. It has been reported that survivin is expressed during fetal development and in cancer tissues.3 Furthermore,survivin overexpression, by disrupting the balance between cell proliferation/differentiation and apoptosis, may relate with the resistance to a variety of apoptotic stimuli, including chemotherapy.4,5 We designed antisense oligonucleotides of survivin to treat the drug-resistant human ovarian carcinoma cell line COC1/DDP, and studied its effects on inducing COC1/DDP apoptosis. The purpose of this study was to find a novel approach to improve the sensitivity of ovarian carcinoma chemotherapy.

  12. Expression and clinical significance of Smac and survivin in esophageal squamous carcinoma%食管鳞癌组织中Smac、survivin的表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    李爱光; 孟宪利; 王洪琰; 刘庆熠; 何明; 王世杰

    2008-01-01

    目的 探讨Smac及survivin在食管鳞癌中的表达变化及其对食管癌预后的判断价值.方法 采用免疫组化方法 测定食管鳞癌组织及癌周正常食管组织中的Smac、survivin.结果 Smac在癌周正常食管组织中阳性表达高于在癌组织中的表达,survivin在癌周正常食管组织中阳性表达低于在癌组织中的表达;Smae在长期生存的食管癌患者中多呈高表达,survivin多呈低表达.Smac与survivin在食管鳞癌中的表达呈负相关(r=-0.288,P=0.013).结论 Smae及survivin可能是食管鳞癌细胞凋亡信号传导网络中的重要因子,是判断食管鳞癌预后的重要指标.

  13. Expression of Survivin in transitional cell carcinoma of urinary bladder and its clinical significance%Survivin蛋白在膀胱移行细胞癌组织中的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    肖泽均; 郑闪; 陈汶; 张海峰; 李长岭; 高燕宁

    2004-01-01

    目的分析Survivin蛋白在膀胱移行细胞癌(TCC)组织中的表达,探讨其潜在的临床意义.方法用免疫组织化学SP法检测75例膀胱TCC患者的肿瘤组织和7份正常膀胱黏膜组织中Survivin蛋白的表达.结果在75例膀胱TCC患者的肿瘤组织中,有58例Survivin蛋白表达阳性,阳性率为77.3%;而在7份正常膀胱黏膜组织中Survivin蛋白均未表达.Survivin蛋白的表达与膀胱TCC的组织学分级密切相关(P0.05).结论凋亡抑制蛋白Survivin 在膀胱TCC患者的肿瘤组织中过度表达,提示Survivin基因在膀胱TCC的发生发展中可能起重要作用;Survivin蛋白的表达与膀胱TCC的组织学分级有明显的相关性,可能成为膀胱TCC不良预后的指标之一.

  14. Berbamine selectively induces apoptosis of human acute promyelocytic leukemia cells via survivin-mediated pathway

    Institute of Scientific and Technical Information of China (English)

    ZHAO Xiao-ying; HE Zhi-wen; WU Dong; XU Rong-zhen

    2007-01-01

    Background Currently, resistance and relapse are still major problems in acute promyelocytic leukemia (APL) cases.Thus, new agents that override the resistance are crucial to the development of curative therapies for APL. In this study,we investigated the effects of berbamine on the proliferation of APL cell line NB4 and its possible mechanisms.Methods NB4 cells were treated with berbamine at different concentrations (0-64 μg/ml) for 72 hours. MTT assay was used to determine proliferation inhibition of NB4 cells. Cell apoptosis was evaluated by both flow cytometry (FCM) and morphological examination. PML/RAR-α and survivin mRNAs were measured by nested-RT-PCR and RT-PCR,respectively. Activated-caspase 3 was determined by FCM.Results Berbamine greatly inhibited the proliferation of NB4 cells in dose- and time-dependent manners, and its IC50 value was 3.86 μg/ml at 48 hours. Both morphological observations and FCM results showed that berbamine induced apoptosis of NB4 cells with concomitant increase of activated caspase-3 and decrease of survivin mRNA. After treatment with berbamine at 8 μg/ml for 48 hours, the percentage of apoptotic cells increased from 2.83% to 58.44% (P<0.01), and the percentage of cells with activated-caspase 3 elevated from 2.06% to 70.89% (P<0.01), whereas, level of survivin mRNA was reduced to 38.24% of control (P<0.01). However, no significant change was observed in PML/RAR-α mRNA.Conclusions Berbamine induces caspase-3-dependent apoptosis of leukemia NB4 cells via survivin-mediated pathway, suggesting that berbamine may be a novel potential agent against APL with a mechanism distinct from that of all-trans retinoic acid (ATRA) and arsenic trioxide (ATO).

  15. Calcium and calcium sensing receptor modulates the expression of thymidylate synthase, NAD(P)H:quinone oxidoreductase 1 and survivin in human colon carcinoma cells: promotion of cytotoxic response to mitomycin C and fluorouracil

    OpenAIRE

    Liu, Guangming; Hu, Xin; Varani, James; Chakrabarty, Subhas

    2009-01-01

    Ca2+ and the cell-surface calcium sensing receptor (CaSR) constitute a novel and robust ligand/receptor system in regulating the proliferation and differentiation of colonic epithelial cells. Here we show that activation of CaSR by extracellular Ca2+ (or CaSR agonists) enhanced the sensitivity of human colon carcinoma cells to mitomycin C (MMC) and fluorouracil (5-FU). Activation of CaSR up-regulated the expression of MMC activating enzyme, NAD(P)H:quinone oxidoreductase 1 (NQO-1) and down-re...

  16. Indomethacin promotes apoptosis in gastric cancer cells through concomitant degradation of Survivin and Aurora B kinase proteins.

    Science.gov (United States)

    Chiou, Shiun-Kwei; Hoa, Neil; Hodges, Amy; Ge, Lishen; Jadus, Martin R

    2014-09-01

    Regular usage of nonsteroidal anti-inflammatory drugs (NSAIDs) is associated with reduced incidence of a variety of cancers. The molecular mechanisms underlying these chemopreventive effects remain poorly understood. This current investigation showed that in gastric cancer cells: (1) Indomethacin treatment enhanced the degradation of chromosomal passenger proteins, Survivin and Aurora B kinase; (2) Indomethacin treatment down-regulated Aurora B kinase activity in a cell cycle-independent fashion; (3) siRNA knockdown of Survivin level promoted Aurora B kinase protein degradation, and vice versa; (4) ectopic overexpression of Survivin blocked reduction of Aurora B kinase level and activity by indomethacin treatment, and vice versa; (5) siRNA knockdown of Aurora B kinase level and AZD1152 inhibition of its activity induced apoptosis, and overexpression of Aurora B kinase inhibited indomethacin-induced apoptosis; (6) indomethacin treatment reduced Aurora B kinase level, coinciding with reduction of Survivin level and induction of apoptosis, in KATO III and HT-29 cells, and in mouse gastric mucosa. A role for Aurora B kinase function in NSAID-induced apoptosis was not previously explored. Thus this report provides better understanding of the molecular mechanisms underlying the anti-cancer effect of NSAIDs by elucidating a significant role for Aurora B kinase in indomethacin-induced apoptosis.

  17. Detection of autoantibodies against survivin in sera from cancer dogs.

    Science.gov (United States)

    Tango, Yumiko; Kano, Rui; Maruyama, Haruhiko; Asano, Kazushi; Tanaka, Shigeo; Hasegawa, Astuhiko; Kamata, Hiroshi

    2010-07-01

    Survivin overexpression has been reported in relation to tumor malignancy, suggesting that it is an unfavorable prognostic marker, and antibody responses to this protein have been confirmed in human cancer patients. In this study, we investigated antibody responses to survivin in canine cancer cases, and examined the prevalence of such responses by enzyme-linked immunosorbent assay (ELISA) using recombinant canine survivin protein as the antigen. The cut-off value for positivity in the anti-survivin ELISA was 0.35, as determined using the mean absorbance +2 S.D. of samples from healthy dogs. Sera from 16 of 59 (27.1%) cancer and 3 of 25 (12%) non-cancer disease dogs were positive on ELISA. The highest positivity rates (>50%) among the cancer cases were seen in dogs with mammary tumor, squamous cell carcinoma and melanoma.

  18. SURVIVIN as a marker for quiescent-breast cancer stem cells-An intermediate, adherent, pre-requisite phase of breast cancer metastasis.

    Science.gov (United States)

    Siddharth, Sumit; Das, Sarita; Nayak, Anmada; Kundu, Chanakya Nath

    2016-10-01

    Cancer stem cells drive the metastatic cascade by undergoing epithelial to mesenchymal transition (EMT) and again mesenchymal to epithelial transition (MET). Using multiple breast cancer cell lines including cigarette smoke induced breast cancer cells and tumor derived primary cells from patient sample; we developed a breast cancer metastasis model and reported the existence of an adherent, distinct pre-metastatic phase, quiescent-breast cancer stem cells (Q-BCSCs) prior to attaining an EMT. SURVIVIN was found to be expressed in Q-BCSCs. Time dependant biphasic expression of SURVIVIN in Q-BCSCs reveals that Q-BCSCs is a pre-metastatic phase distinct from both epithelial and mesenchymal counterparts. SURVIVIN favours metastasis and up-regulates WNT/β-CATENIN pathway in a PI3 K/AKT-dependant manner for self-renewal. Knockdown of SURVIVIN in Q-BCSCs lost the metastatic property of cells by inhibiting invasion, EMT-MET, PI3 K/AKT/WNT cascade, and induced apoptosis. Thus, our data suggest the existence of a novel pre-metastatic phase (Q-BCSCs) before EMT and SURVIVIN acts as a marker for Quiescent-BCSCs.

  19. Inhibitor of Apoptosis (IAP) survivin is indispensable for survival of HER2 gene-amplified breast cancer cells with primary resistance to HER1/2-targeted therapies

    Energy Technology Data Exchange (ETDEWEB)

    Oliveras-Ferraros, Cristina; Vazquez-Martin, Alejandro; Cufi, Silvia; Torres-Garcia, Violeta Zenobia [Unit of Translational Research, Catalan Institute of Oncology-Girona, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Girona Biomedical Research Institute, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Sauri-Nadal, Tamara; Barco, Sonia Del [Girona Biomedical Research Institute, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Medical Oncology, Catalan Institute of Oncology-Girona, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Lopez-Bonet, Eugeni [Girona Biomedical Research Institute, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Department of Anatomical Pathology, Dr. Josep Trueta University Hospital, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Brunet, Joan [Girona Biomedical Research Institute, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Medical Oncology, Catalan Institute of Oncology-Girona, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Martin-Castillo, Begona [Girona Biomedical Research Institute, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Unit of Clinical Research, Catalan Institute of Oncology-Girona, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Menendez, Javier A., E-mail: jmenendez@idibgi.org [Unit of Translational Research, Catalan Institute of Oncology-Girona, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain); Girona Biomedical Research Institute, Avenida de Francia S/N, E-17007 Girona, Catalonia (Spain)

    2011-04-08

    Highlights: {yields} Intrinsic trastuzumab resistance occurs in {approx}70% of metastatic HER2 + breast carcinomas (BC). {yields} Approximately 15% of early HER2 + BC relapse in spite of treatment with trastuzumab-based therapies. {yields} HER2-independent downstream pro-survival pathways might underlie trastuzumab refractoriness. {yields} Survivin is indispensable for proliferation and survival of HER2 + BC unresponsive to HER2-targeted therapies ab initio. {yields} Survivin antagonists may clinically circumvent the occurrence of de novo resistance to HER2-directed drugs. -- Abstract: Primary resistance of HER2 gene-amplified breast carcinomas (BC) to HER-targeted therapies can be explained in terms of overactive HER2-independent downstream pro-survival pathways. We here confirm that constitutive overexpression of Inhibitor of Apoptosis (IAP) survivin is indispensable for survival of HER2-positive BC cells with intrinsic cross-resistance to multiple HER1/2 inhibitors. The IC{sub 50} values for the HER1/2 Tyrosine Kinase Inhibitors (TKIs) gefitinib, erlotinib and lapatinib were up to 40-fold higher in trastuzumab-unresponsive JIMT-1 cells than in trastuzumab-naive SKBR3 cells. ELISA-based and immunoblotting assays demonstrated that trastuzumab-refractory JIMT-1 cells constitutively expressed {approx}4 times more survivin protein than trastuzumab-responsive SKBR3 cells. In response to trastuzumab, JIMT-1 cells accumulated {approx}10 times more survivin than SKBR3 cells. HER1/2 TKIs failed to down-regulate survivin expression in JIMT-1 cells whereas equimolar doses of HER1/HER2 TKIs drastically depleted survivin protein in SKBR3 cells. ELISA-based detection of histone-associated DNA fragments confirmed that trastuzumab-refractory JIMT-1 cells were intrinsically protected against the apoptotic effects of HER1/2 TKIs. Of note, when we knocked-down survivin expression using siRNA and then added trastuzumab, cell proliferation and colony formation were completely

  20. Expression of Intratumoral IGF-II Is Regulated by the Gene Imprinting Status in Triple Negative Breast Cancer from Vietnamese Patients

    Directory of Open Access Journals (Sweden)

    Vinodh Kumar Radhakrishnan

    2015-01-01

    Full Text Available African American women suffer higher incidence and mortality of triple negative breast cancer (TNBC than Caucasian women. TNBC is very aggressive, causing the worst clinical outcome. We previously demonstrated that tumors from these patients express high IGF-II and exhibit high activation of the IGF signaling pathways. IGF-II gene expression is imprinted (monoallelic, promotes tumor progression, and metastasis and regulates Survivin, a TNBC prognostic marker. Since BC mortality has increased among young Vietnamese women, we analyzed 48 (paired TNBC samples from Vietnamese patients to assess IGF-II expression. We analyzed all samples by qrtPCR for identification of IGF-II heterozygosity and to determine allelic expression of the IGF-II gene. We also analyzed the tissues for proIGF-II and Survivin by RT-PCR and Western blotting. A total of 28 samples displayed IGF-II heterozygosity of which 78% were biallelic. Tumors with biallelic IGF-II gene expression exhibited the highest levels of proIGF-II and Survivin. Although 100% of these tissues corresponding normal samples were biallelic, they expressed significantly lower levels of or no proIGF-II and Survivin. Thus, IGF-II biallelic gene expression is differentially regulated in normal versus tumor tissues. We propose that intratumoral proIGF-II is dependent on the IGF-II gene imprinting status and it will promote a more aggressive TNBC.

  1. Rapamycin enhances docetaxel-induced cytotoxicity in a androgen-independent prostate cancer xenograft model by survivin downregulation

    Energy Technology Data Exchange (ETDEWEB)

    Morikawa, Yasuyuki, E-mail: yasu-m@med.gunma-u.ac.jp [Department of Urology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maeabshi, Gunma 3718511 (Japan); Koike, Hidekazu; Sekine, Yoshitaka; Matsui, Hiroshi; Shibata, Yasuhiro; Ito, Kazuto; Suzuki, Kazuhiro [Department of Urology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maeabshi, Gunma 3718511 (Japan)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer Rapamycin (RPM) enhances the susceptibility of PC3 cells to docetaxel. Black-Right-Pointing-Pointer Low-dosage of docetaxel (DTX) did not reduce survivin expression levels in PC3 cells. Black-Right-Pointing-Pointer Combination treatment of RPM with DTX suppressed the expression of surviving. Black-Right-Pointing-Pointer SiRNA against survivin enhanced the susceptibility of PC3 cells to DTX. Black-Right-Pointing-Pointer RPM and DTX cotreatment inhibited PC3 cell growth and decreased surviving in vivo. -- Abstract: Background: Docetaxel is a first-line treatment choice in castration-resistant prostate cancer (CRPC). However, the management of CRPC remains an important challenge in oncology. There have been many reports on the effects of rapamycin, which is an inhibitor of the mammalian target of rapamycin (mTOR), in the treatment of carcinogenesis. We assessed the cytotoxic effects of the combination treatment of docetaxel and rapamycin in prostate cancer cells. Furthermore, we examined the relationship between these treatments and survivin, which is a member of the inhibitory apoptosis family. Methods: Prostate cancer cells were cultured and treated with docetaxel and rapamycin. The effects on proliferation were evaluated with the MTS assay. In addition, we evaluated the effect on proliferation of the combination treatment induced knockdown of survivin expression by small interfering RNA transfection and docetaxel. Protein expression levels were assayed using western blotting. PC3 cells and xenograft growth in nude mice were used to evaluate the in vivo efficacy of docetaxel and its combination with rapamycin. Results: In vitro and in vivo, the combination of rapamycin with docetaxel resulted in a greater inhibition of proliferation than treatment with rapamycin or docetaxel alone. In addition, in vitro and in vivo, rapamycin decreased basal surviving levels, and cotreatment with docetaxel further decreased these levels

  2. Sulindac sulfide induces autophagic death in gastric epithelial cells via survivin down-regulation: a mechanism of NSAIDs-induced gastric injury.

    Science.gov (United States)

    Chiou, Shiun-Kwei; Hoa, Neil; Hodges, Amy

    2011-06-01

    Sulindac sulfide, a nonsteroidal anti-inflammatory drug (NSAID), has anti-tumorigenic and anti-inflammatory activities, but causes gastric mucosal damage. NSAIDs cause gastric injury in part by down-regulation of Survivin, an apoptosis inhibitor, resulting in apoptosis induction. Autophagy is a process that promotes cellular health by destroying unwanted cellular materials. Excessive autophagy induction could lead to a non-apoptotic cell death (autophagic cell death). The present study showed that sulindac sulfide at a physiological concentration also induces autophagic death in human gastric epithelial AGS and rat gastric epithelial RGM-1 cells, and that Survivin down-regulation is a mechanism involved: Sulindac sulfide treatment increased LC3b-II and APG7 levels and cytosolic vacuole formation, indications of autophagy induction, in AGS and RGM-1 cells. Sulindac sulfide treatment induced AGS and RGM-1 cell death, which was significantly reduced by pretreatment with the autophagy inhibitors 3-methyladenine and chloroquine, indicating that sulindac sulfide induced autophagic cell death. Stable overexpression of Survivin in RGM-1 cells did not inhibit the induction of LC3b-II levels or vacuole formation by sulindac sulfide, but significantly reduced the resulting cell death, suggesting that Survivin may inhibit autophagic cell death downstream of LC3b-II induction and vacuole formation. Indeed, siRNA depletion of LC3b in AGS cells inhibited the down-regulation of Survivin levels and the induction of cell death by sulindac sulfide, confirming that down-regulation of Survivin occurs in the autophagy pathway downstream of LC3b-II induction by sulindac sulfide. Induction of Survivin-dependent autophagic cell death is a novel mechanism by which sulindac sulfide induces gastric mucosal injury.

  3. Effect of microRNA-494 over-expression and RNA interference-mediated Survivin gene inhibition on xenograft prostate cancer growth%微小RNA-494过表达与RNA干扰抑制Survivin基因对前列腺癌移植瘤生长的比较

    Institute of Scientific and Technical Information of China (English)

    孙承文; 王礼平; 臧亚晨; 薛波新; 单玉喜; 许立军; 阳东荣

    2013-01-01

    Objective To investigate the effect of microRNA-494 over-expression and RNA interference-mediated Survivin gene inhibition on on xenograft prostate cancer growth in nude mice.Methods Two adenovirns vectors,Ad-494 which can overexpressed microRNA-494,and Ad-sur loaded with Survivin short hairpin RNA,were transfected into PC-3 cells alone or in combination,and the following groups were established:PBS group (blank control),Ad-GFP group (negative control),Ad-sur group,Ad-494 group,Ad-sur + Ad-494 group.Then a total of 1 × 107 transfected cells were injected to the right armpit of one nude mouse to establish xenograft prostate cancer model.The gross tumor volumes were measured periodically.Fifty-five days after cell injection,the mice were sacrificed and tumor volume was measured.The expression of Survivin gene in tumor sample was detected by using Western blotting and immunohistochemistry.The expression of B lymphocytes/leukemia-2 (bcl-2),bcl-2 associated X protein (bax),and cysteinyl aspartate-specific protease (Caspase)-3 in tumor samples was examined by using immunohistochemistry.Results The tumor growth was inhibited dramatically in Ad-sur group,Ad-494 group and Ad-sur + Ad-494 group as compared with control groups (P < 0.05).Thirty-five days after tumor cell injection,the difference in the tumor volume was found in each group.The average volume in Ad-sur + Ad-494 group was the smallest (40.69 ±0.69) mm3 as compared to that in either Ad-sur group (102.11 ± 5.32) mm3 or Ad-494 group (99.03 ±3.50) mm3(P < 0.05).Fifty-five days after tumor cell injection,the tumor inhibition rate in Ad-sur group,Ad-494 group and Ad-sur + Ad-494 group was 64.62%,65.98% and 86.67%,respectively,suggesting the synergetic anti-tumor effect of Ad-sur + Ad-494 (Q =0.99).Both Western blotting and immunohistochemistry revealed that Survivin gene expression was down-regulated in all experimental groups as compared with control groups,more obviously in Ad-sur + Ad-494 group (P

  4. Resveratrol down-regulates survivin and induces apoptosis in human multidrug-resistant SPC-A-1/CDDP cells.

    Science.gov (United States)

    Zhao, Weiguo; Bao, Pengtao; Qi, Haowen; You, Houcheng

    2010-01-01

    We studied the effect of resveratrol treatment on multidrug-resistant human non-small cell lung cancer cells. Human multidrug-resistant SPC-A-1/CDDP cells were treated with resveratrol at a concentration of 25, 50, or 100 microM in in vitro studies and nude mice were implanted with multidrug-resistant SPC-A-1/and fed a special diet that included resveratrol at a dose of either 1 g/kg/day or 3 g/kg/day in in vivo studies. No adverse toxicological effects of resveratrol treatment were observed. The rate of cell proliferation, apoptosis ratio, cell cycle phase distribution, IC50 values of cisplatin, gefitinib, and paclitaxel, implanted tumour volume, and expression of survivin in resveratrol-treated and control mice were then determined. Resveratrol significantly inhibited the proliferation of SPC-A-1/CDDP cells, induced apoptosis, arrested the cell cycle phase between G0-G1 and S phase or at the G2/M phase, decreased the IC50 values of multiple chemotherapeutic drugs, and showed anti-tumour effects in nude mice that had been implanted with SPC-A-1/CDDP cells. In additional, resveratrol affected the proliferation of SPC-A-1/CDDP cells in a dose- and time-dependent manner. Expression of survivin in SPC-A-1/CDDP cells decreased after they were treated with all concentrations of resveratrol and resveratrol was also found to have a dose-dependent effect on survivin expression. Resveratrol can induce apoptosis in multidrug-resistant human NSCLC SPC-A-1/CDDP cells by down-regulating the expression of survivin.

  5. Survivin promoter-regulated oncolytic adenovirus with Hsp70 gene exerts effective antitumor efficacy in gastric cancer immunotherapy

    OpenAIRE

    Wang, Weiguo; Ji, Weidan; Hu, Huanzhang; Ma, Juming; Li, Xiaoya; Mei, Weiqun; Xu, Yang; Hu, Huizhen; Yan, Yan; Song, Qizhe; Li, Zhigang; Su, Changqing

    2013-01-01

    Gene therapy is a promising adjuvant therapeutic strategy for cancer treatment. To overcome the limitations of current gene therapy, such as poor transfection efficiency of vectors, low levels of transgene expression and lack of tumor targeting, the Survivin promoter was used to regulate the selective replication of oncolytic adenovirus in tumor cells, and the heat shock protein 70 (Hsp70) gene was loaded as the anticancer transgene to generate an AdSurp-Hsp70 viral therapy system. The effica...

  6. Survivin promoter-regulated oncolytic adenovirus with Hsp70 gene exerts effective antitumor efficacy in gastric cancer immunotherapy.

    Science.gov (United States)

    Wang, Weiguo; Ji, Weidan; Hu, Huanzhang; Ma, Juming; Li, Xiaoya; Mei, Weiqun; Xu, Yang; Hu, Huizhen; Yan, Yan; Song, Qizhe; Li, Zhigang; Su, Changqing

    2014-01-15

    Gene therapy is a promising adjuvant therapeutic strategy for cancer treatment. To overcome the limitations of current gene therapy, such as poor transfection efficiency of vectors, low levels of transgene expression and lack of tumor targeting, the Survivin promoter was used to regulate the selective replication of oncolytic adenovirus in tumor cells, and the heat shock protein 70 (Hsp70) gene was loaded as the anticancer transgene to generate an AdSurp-Hsp70 viral therapy system. The efficacy of this targeted immunotherapy was examined in gastric cancer. The experiments showed that the oncolytic adenovirus can selectively replicate in and lyse the Survivin-positive gastric cancer cells, without significant toxicity to normal cells. AdSurp-Hsp70 reduced viability of cancer cells and inhibited tumor growth of gastric cancer xenografts in immuno-deficient and immuno-reconstruction mouse models. AdSurp-Hsp70 produced dual antitumor effects due to viral replication and high Hsp70 expression. This therapeutic system used the Survivin promoter-regulated oncolytic adenovirus vector to mediate targeted expression of the Hsp70 gene and ensure safety and efficacy for subsequent gene therapy programs against a variety of cancers.

  7. Differential regulation of survivin by p53 contributes to cell cycle dependent apoptosis

    Institute of Scientific and Technical Information of China (English)

    Yan JIN; Yong WEI; Lei XIONG; Ying YANG; Jia Rui WU

    2005-01-01

    Recent studies indicate that cell-cycle checkpoints are tightly correlated with the regulation of apoptosis, in which p53 plays an important role. Our present works show that the expression of E6/E7 oncogenes of human papillomavirus in HeLa cells is inhibited in the presence of anti-tumor reagent tripchlorolide (TC), which results in the up-regulation of p53 in HeLa cells. Interestingly, under the same TC-treatment, the cells at the early S-phase are more susceptible to apoptosis than those at the middle S-phase although p53 protein is stabilized to the same level in both situations.Significant difference is exhibited between the two specified expression profiles. Further analysis demonstrates that anti-apoptotic gene survivin is up-regulated by p53 in the TC-treated middle-S cells, whereas it is down-regulated by p53 in the TC-treated early-S cells. Taken together, the present study indicates that the differential p53-regulated expression of survivin at different stages of the cell cycle results in different cellular outputs under the same apoptosis-inducer.

  8. Enhancement of survivin gene downregulation and cell apoptosis by a novel combination: liposome microbubbles and ultrasound exposure.

    Science.gov (United States)

    Chen, Zhiyi; Liang, Kun; Liu, Jianhua; Xie, Mingxing; Wang, Xinfang; Lü, Qing; Zhang, Jing; Fang, Lingyun

    2009-12-01

    Ultrasound-mediated microbubble destruction (sonoporation) is an efficient and safe nonviral technique for gene delivery. In the present work, we hypothesized that short hairpin RNA (shRNA) interference therapy targeting human Survivin gene could be transfected by the novel combination of ultrasound exposure (USE) and liposome microbubbles (LM). ShRNA vectors targeting Survivin were constructed and transfected under USE and LM conditions. The optimal transfection efficiency and cell injury were compared with those of polyethylenimine (PEI)-mediated transfection in different cancer cell lines (HeLa, HepG2, Ishikawa, MCF-7, and B16-F10). The effects of gene downregulation and cell apoptosis were further investigated. The results indicated that P + USE + LM group could significantly increase the gene expression as compared with plasmid group, plasmid + USE group, plasmid + LM group (P < 0.001). The transfection efficiency of the novel combination was nearly equal to PEI-mediated transfection in some cancer cell lines while the cell viability did not decrease markedly. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis also confirmed that Survivin mRNA and protein expression could be knocked down significantly by shRNA transfection under USE and LM condition (P < 0.001). This is the first study to verify the role of shRNA therapy in vitro with novel combination of USE and LM. We concluded that this nonviral technique would be valuable in the gene transfection of shRNA and Survivin gene downregulation would lead to apparent cell apoptosis.

  9. Clinical and immunological evaluation of anti-apoptosis protein, survivin-derived peptide vaccine in phase I clinical study for patients with advanced or recurrent breast cancer

    Directory of Open Access Journals (Sweden)

    Asanuma Hiroko

    2008-05-01

    Full Text Available Abstract Background We previously reported that survivin-2B, a splicing variant of survivin, was expressed in various types of tumors and that survivin-2B peptide might serve as a potent immunogenic cancer vaccine. The objective of this study was to examine the toxicity of and to clinically and immunologically evaluate survivin-2B peptide in a phase I clinical study for patients with advanced or recurrent breast cancer. Methods We set up two protocols. In the first protocol, 10 patients were vaccinated with escalating doses (0.1–1.0 mg of survivin-2B peptide alone 4 times every 2 weeks. In the second protocol, 4 patients were vaccinated with the peptide at a dose of 1.0 mg mixed with IFA 4 times every 2 weeks. Results In the first protocol, no adverse events were observed during or after vaccination. In the second protocol, two patients had induration at the injection site. One patient had general malaise (grade 1, and another had general malaise (grade 1 and fever (grade 1. Peptide vaccination was well tolerated in all patients. In the first protocol, tumor marker levels increased in 8 patients, slightly decreased in 1 patient and were within the normal range during this clinical trial in 1 patient. With regard to tumor size, two patients were considered to have stable disease (SD. Immunologically, in 3 of the 10 patients (30%, an increase of the peptide-specific CTL frequency was detected. In the second protocol, an increase of the peptide-specific CTL frequency was detected in all 4 patients (100%, although there were no significant beneficial clinical responses. ELISPOT assay showed peptide-specific IFN-γ responses in 2 patients in whom the peptide-specific CTL frequency in tetramer staining also was increased in both protocols. Conclusion This phase I clinical study revealed that survivin-2B peptide vaccination was well tolerated. The vaccination with survivin-2B peptide mixed with IFA increased the frequency of peptide-specific CTL more

  10. Decitabine and SAHA-Induced Apoptosis Is Accompanied by Survivin Downregulation and Potentiated by ATRA in p53-Deficient Cells

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    Barbora Brodská

    2014-01-01

    Full Text Available While p53-dependent apoptosis is triggered by combination of methyltransferase inhibitor decitabine (DAC and histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA in leukemic cell line CML-T1, reactive oxygen species (ROS generation as well as survivin and Bcl-2 deregulation participated in DAC + SAHA-induced apoptosis in p53-deficient HL-60 cell line. Moreover, decrease of survivin expression level is accompanied by its delocalization from centromere-related position in mitotic cells suggesting that both antiapoptotic and cell cycle regulation roles of survivin are affected by DAC + SAHA action. Addition of subtoxic concentration of all-trans-retinoic acid (ATRA increases the efficiency of DAC + SAHA combination on viability, apoptosis induction, and ROS generation in HL-60 cells but has no effect in CML-T1 cell line. Peripheral blood lymphocytes from healthy donors showed no damage induced by DAC + SAHA + ATRA combination. Therefore, combination of ATRA with DAC and SAHA represents promising tool for therapy of leukemic disease with nonfunctional p53 signalization.

  11. Analysis of polymorphism in the survivin gene promoter as a potential risk factor for head and neck cancers development

    Directory of Open Access Journals (Sweden)

    Kostić Marija

    2013-01-01

    Full Text Available Introduction. Association studies have shown that gene polymorphisms in various classes of genes can modulate cancer risk. The -31G/C polymorphism in the promoter of survivin gene, affects the expression of the anti-apoptotic protein survivin which in turn may predispose an individual to some types of cancer. Objective. The aim of the study was to determine whether the survivin promoter -31G/C polymorphism could be a susceptibility factor for squamous cell carcinoma (SCC of the oral cavity and basal cell carcinoma (BCC of the skin. Methods. The DNA obtained from 88 patients with SCC, 60 patients with BCC and 111 healthy individuals was subjected to polymerase chain reaction-restriction fragment length polymorphism analysis (PCR- RFLP in order to determine genotype and allele frequencies in patients and control groups. Logistic regression was used for cancer risk assessment. Results. The following distribution of genotypes was obtained: CC genotype 15% in the SCC group, 13% in the BCC group and 12% in controls; CG genotype 41% in SCCs, 35% in BCCs, 48% in controls; GG genotype 44% in SCCs, 52% in BCCs and 40% in controls. Allelic frequencies were as follows: G allele 0.65 in SCCs, 0.69 in BCCs and 0.64 in the control group; C allele 0.35 in SCCs, 0.31 in BCCs and 0.36 in the control group. There was no statistically significant difference in allele or genotype frequencies between the patients and controls (p>0.05. Conclusion. In Serbian population, -31G/C polymorphism in the promoter of the survivin gene cannot be considered as a risk factor for oral squamous cell carcinoma and skin basal cell carcinoma. [Projekat Ministarstva nauke Republike Srbije, br. 175075

  12. CD44 Influences Fibroblast Behaviors Via Modulation of Cell-Cell and Cell-Matrix Interactions, Affecting Survivin and Hippo Pathways.

    Science.gov (United States)

    Tsuneki, Masayuki; Madri, Joseph A

    2016-03-01

    CD44 has been studied in a wide variety of cell types, in a diverse array of cell behaviors and in a diverse range of signaling pathways. We now document a role for CD44 in mediating fibroblast behaviors via regulation of N-cadherin, extracellular matrix expression, Survivin and the Hippo pathway. Here, we report our findings on the roles of CD44 in modulating proliferation, apoptosis, migration and invasion of murine wild-type (WT-FB) and CD44 knockout dermal fibroblasts (CD44KO-FB). As we have documented in microvascular endothelial cells lacking CD44, we found persistent increased proliferation, reduced activation of cleaved caspase 3, increased initial attachment, but decreased strength of cell attachment in high cell density, post confluent CD44KO-FB cultures. Additionally, we found that siRNA knock-down of CD44 mimicked the behaviors of CD44KO-FB, restoring the decreases in N-cadherin, collagen type I, fibronectin, Survivin, nuclear fractions of YAP and phospho-YAP and decreased levels of cleaved caspase 3 to the levels observed in CD44KO-FB. Interestingly, plating CD44KO-FB on collagen type I or fibronectin resulted in significant decreases in secondary proliferation rates compared to plating cells on non-coated dishes, consistent with increased cell adhesion compared to their effects on WT-FB. Lastly, siRNA knockdown of CD44 in WT-FB resulted in increased fibroblast migration compared to WT-FB, albeit at reduced rates compared to CD44KO-FB. These results are consistent with CD44's pivotal role in modulating several diverse behaviors important for adhesion, proliferation, apoptosis, migration and invasion during development, growth, repair, maintenance and regression of a wide variety of mesenchymal tissues.

  13. The role of survivin in the pathogenesis of pulmonary hypertension%Survivin在肺动脉高压发病过程中的作用

    Institute of Scientific and Technical Information of China (English)

    张帅; 樊再雯

    2014-01-01

    Survivin是凋亡抑制蛋白家族的新成员,也是目前发现的最强的凋亡抑制蛋白.Survivin参与调控细胞周期的两个基本过程,即抑制细胞凋亡和促进细胞增殖.目前研究表明survivin调节肺动脉平滑肌细胞的增殖,在肺动脉高压的肺动脉中膜高表达.通过研究survivin在肺动脉高压发病过程中的作用,可以为肺动脉高压的治疗提供新的思路和手段.%Survivin is a new member of the inhibitor of apoptosis (IAP) family of proteins,which is also currently the strongest discovered inhibitor of apoptosis protein.Survivin regulates two essential cellular processes,i.e.,it inhibits apoptosis and promotes cell proliferation.Recent studies show that survivin regulates pulmonary artery smooth muscle cell proliferation,which keeps on a high level expression in media of pulmonary artery from pulmonary hypertension.By studying in the role of survivin in the pathogenesis of pulmonary hypertension can provide new ideas and means in therapy of pulmonary hypertension.

  14. 冬凌草甲素和survivin反义核苷酸对前列腺癌细胞作用的研究%Effects of survivin antisense oligodeoxynecleotides and Oridonin on PC-3 cells

    Institute of Scientific and Technical Information of China (English)

    李进; 杨罗艳; 吴洪涛

    2014-01-01

    Objective To explore the synergistic effects of survivin antisense oligonucleotides combined with Oridonin on growth, apoptosis, and the expression of survivin of PC-3 cells. Methods Human prostate carcinoma cells PC-3 on logarithmic growth phase were used in this study. The cell vitality was determined by MTT assay. The combination index (CI) was calculated using Pharmaconamics CalcuSynsoftware. The apoptotic rate was examined by flow cytometer (FCM). The expression of survivin was detected by Western Blot and Real-time Fluorescent Quantitation-PCR. Results After transfection with antisense Survivin RNAi, the proliferation of PC-3 cells was inhibited markedly. An obvious apoptosis was found in the transfected PC-3 cells. The inhibitory effect of combined administration of survivin antisense and Oridonin on cell proliferation was much stronger than that of the single way (P<0.01). It showed that there was a synergistic effect (Fa<0.80). Western Blot and RT-PCR assays demonstrated that survivin antisense and Oridonin all inhibited the expression of survivin(P <0.01). Conclusion Combined survivin antisense and Oridonin significantly inhibits cell proliferation, induces cell apoptosis and down-regulates survivin expression in PC-3 cells, indicating that survivin antisense and Oridonin have a synergistic effect on PC-3 cells.%目的:探讨冬凌草甲素联合survivin反义核苷酸(反义链)对前列腺癌PC-3细胞株增殖和凋亡以及survivin mRNA和蛋白的影响。方法常规培养PC-3细胞,用四甲基偶氮唑盐法(MTT法)检测survivin反义链联合冬凌草甲素对PC-3细胞增殖的影响;流式细胞仪(FCM)检测PC-3细胞凋亡率;以CalcuSyn药效学软件计算联合指数(CI)评价survivin反义链联合凌草甲素对PC-3细胞的联合效应,并通过荧光定量PCR和Western blot方法检测PC-3细胞survivin基因和蛋白表达变化。结果 survivin反义链转染PC-3细胞后,可以显著抑制PC-3

  15. Promoter methylation of survivin gene in infantile hemangiomas%婴幼儿血管瘤survivin基因启动子甲基化的研究

    Institute of Scientific and Technical Information of China (English)

    熊耕; 刘铭; 彭强; 植勇; 熊小明

    2016-01-01

    infantile hemangiomas and normal skin tissues by bisulfite sequencing PCR (BSP) and examine the possible relationship of promoter methylation in survivin gene and the proliferation or involution of infantile hemangiomas.Methods The immunohistochemical method of S-P was used for detecting the expression of survivin protein in paraffin specimens of infantile proliferative hemangioma (n =30),involuting hemangioma (n =30) and normal foreskin tissues (n =10);tissue genomic DNA was extracted from paeaffin embedding.After purification,the method of BSP was employed for detecting the methylation status of CpG island in survivin gene promoter region of proliferative hemangioma (n =30),involuting hemangioma (n =30) and normal foreskin tissues (n =10).Results The positive expression rates of survivin protein in proliferative hemangiomas involuting hemangiomas and normal foreskin tissues were 76.6% (23/30),33.3% (10/30) and 20% (2/10) respectively.Among 10 cases of normal foreskin tissues,only 1 case (10%) of CpG island was demethylated in survivin gene promoter while 9 cases (90%) were methylated;among 30 cases of involuting hemangiomas,8 cases (26.6%) of CpG island were demethylated in survivin gene promoter while 22 cases (73.3%) were methylated;among 30 cases of proliferative hemangiomas,24 cases (80.0%) of CpG island were demethylated in survivin gene promoter and 6 cases (20%) were methylated;The demethylated rate of CpG island in survivin gene promoter was significantly lower in proliferative hemangiomas than those in involuting hemangiomas and normal foreskin tissues.CpG island in survivin gene promoter non-methylated expressed survivin protein in 31/33 hemangiomas while CpG island in survivin gene promoter methylated did not express survivin protein in 26/27 hemangiomas.Conclusions The expression of survivin protein is significantly greater in proliferative hemangiomas than that in involuting hemangiomas.The methylation status of CpG island in survivin gene

  16. Recognizing rheumatoid arthritis: oncoprotein survivin opens new possibilities: a population-based case-control study.

    Science.gov (United States)

    Chun-Lai, Too; Murad, Shahnaz; Erlandsson, Malin C; Hussein, Heselynn; Sulaiman, Wahinuddin; Dhaliwal, Jasbir S; Bokarewa, Maria I

    2015-01-01

    Survivin is a biomarker of cancer known for its anti-apoptotic and cell-cycle regulating properties. In the context of non-cancer pathology, high levels of survivin may be measured in blood and synovial fluid of patients with rheumatoid arthritis (RA) and associate with early joint damage and poor therapy response. The aim of the study was to investigate the value of survivin measurements in blood for diagnosis of RA in the frame of the Malaysian epidemiological investigation of rheumatoid arthritis (MyEIRA) study. The study enrolled RA patients from eight rheumatology centres in Peninsular Malaysia. The healthy controls matched by age, gender and ethnicity were recruited on the community basis from the residential area of the patients. Levels of survivin were measured in blood of RA patients (n = 1233) and controls (n = 1566) by an enzyme-linked immuno-sorbent assay (ELISA). The risk for RA was calculated as odds ratio (OR) and 95% confidence intervals in the individuals with high levels of survivin. The risk was calculated in relation to antibodies against cyclic citrullinated peptides (ACPA), detected by ELISA and HLA-DRB1 shared epitope (SE) alleles, identified by the polymerase chain reaction using sequence specific oligonucleotide method. High levels of survivin were detected in 625 of 1233 (50.7%) RA cases and in 85 of 1566 (5.4%) controls, indicating its high specificity for RA. Survivin was association with an increase in RA risk in the patients having neither SE-alleles nor ACPA (OR = 5.40, 95% CI 3.81-7.66). For the patients combining survivin, SE, and ACPA, the estimated risk for RA was 16-folds higher compared to the survivin negative patients with SE and ACPA(OR = 16.21, 95% CI 5.70-46.18). To conclude, detection of survivin in blood provides a simple test to improve diagnostic and to increase predictability for RA.

  17. Expression of regulators of mitotic fidelity are associated with intercellular heterogeneity and chromosomal instability in primary breast cancer

    DEFF Research Database (Denmark)

    Roylance, Rebecca; Endesfelder, David; Jamal-Hanjani, Mariam

    2014-01-01

    , as determined by centromeric FISH and defined by modal centromere deviation, was analysed. Significantly poorer clinical outcome was observed in patients with high AURKA expression levels. Expression of SURVIVIN was elevated in ER-negative relative to ER-positive breast cancer. Both AURKA and SURVIVIN increased...

  18. Relationship Between Expression of Stathmin, Survivin and Chemotherapy Effect of Docetaxel in Advanced Non-small Cell Lung Cancer%Stathmin及Survivin在非小细胞肺癌中的表达与多西紫杉醇化疗疗效的相关性分析

    Institute of Scientific and Technical Information of China (English)

    原少斐; 朱林佳; 郑维锷

    2015-01-01

    目的 探讨晚期非小细胞肺癌中Stathmin和Survivin的表达与多西紫杉醇化疗疗效的关系.方法 回顾性分析72例接受多西紫杉醇化疗的晚期非小细胞肺癌患者的临床病理资料.免疫组织化学法检测肿瘤标本Stathmin和Survivin蛋白的表达,并对疗效、不良反应进行分析.结果 Stathmin阳性表达率为65.27%(47/72),Survivin阳性表达率为76.38%(55/72).化疗有效率(CR+PR)为51.38%,Stathmin(+)的患者有效率(33.33%)显著低于Stathmin(-)患者(66.67%)(P<0.05).Survivin(+)组的有效率(38.18%)显著低于Survivin(-)组的76.47%(P<0.05).联合检测显示:Stathmin(+)且Survivin(+)组有效率为28.94%,中位疾病无进展时间为3.7个月,1年生存率为29.9%,Stathmin(-)且Survivin(-)组有效率为87.50%,中位疾病无进展时间为7.1个月,1年生存率为62.5%,差异均有显著性(P<0.05).最常见的不良反应为骨髓抑制、消化道反应和脱发.结论 联合检测Stathmin和Survivin的表达可作为多西紫杉醇化疗方案治疗晚期非小细胞肺癌的疗效预测指标之一.

  19. Activating transcription factor 4 mediates a multidrug resistance phenotype of esophageal squamous cell carcinoma cells through transactivation of STAT3 expression.

    Science.gov (United States)

    Zhu, Hongwu; Chen, Xiong; Chen, Bin; Chen, Bei; Fan, Jianyong; Song, Weibing; Xie, Ziying; Jiang, Dan; Li, Qiuqiong; Zhou, Meihua; Sun, Dayong; Zhao, Yagang

    2014-11-01

    Multidrug resistance (MDR) is a major challenge to the clinical treatment of esophageal cancer. The stress response gene activating transcription factor 4 (ATF4) is involved in homeostasis and cellular protection. However, relatively little is known about the expression and function of ATF4 in esophageal squamous cell carcinoma (ESCC) MDR. In this study, we investigate the potential role and mechanisms of ATF4 in ESCC MDR. We demonstrated that overexpression of ATF4 promotes the MDR phenotype in ESCC cells, while depletion of ATF4 in the MDR ESCC cell line induces drug re-sensitization. We also demonstrated that ATF4 transactivates STAT3 expression by directly binding to the signal transducers and activators of transcription 3 (STAT3) promoter, resulting in MDR in ESCC cells. Significantly, inhibition of STAT3 by small interfering RNA (siRNA) or a selective inhibitor (JSI-124) reintroduces therapeutic sensitivity. In addition, increased Bcl-2, survivin, and MRP1 expression levels were observed in ATF4-overexpressing cells. In conclusion, ATF4 may promote MDR in ESCC cells through the up-regulation of STAT3 expression, and thus is an attractive therapeutic target to combat therapeutic resistance in ESCC.

  20. Design, synthesis and biological studies of survivin dimerization modulators that prolong mitotic cycle.

    Science.gov (United States)

    Chettiar, Somsundaram N; Cooley, James V; Park, In-Hee; Bhasin, Deepak; Chakravarti, Arnab; Li, Pui-Kai; Li, Chenglong; Jacob, Naduparambil Korah

    2013-10-01

    Survivin, a member of the inhibitor of apoptosis protein (IAP) family proteins, has essential roles in cell division and inhibition of apoptosis. Several clinical studies in cancer patients have shown that the elevated levels of survivin correlate with aggressiveness of the disease and resistance to radiation and chemotherapeutic treatments. Survivin is an integral component of chromosomal passenger complex (CPC) where it binds to borealin and INCENP through its dimerization interface. Thus, disruption of functional survivin along its dimer interface with a small molecule is hypothesized to inhibit the proliferation of cancer cells and sensitize them to therapeutic agents and radiation. Recently, a small molecule (Abbott8) was reported to bind at the dimerization interface of survivin. Further development of this compound was accomplished by computational modeling of the molecular interactions along the dimerization interface, which has led to the design of promising survivin dimerization modulators. Two of the most potent survivin modulators, LLP3 and LLP9 at concentrations between 50 and 100nM, caused delay in mitotic progression and major mitotic defects in proliferating human umbilical vein endothelial cells (HUVEC) and prostate cancer cells (PC3).

  1. Study on the apoptosis of Raji cell line induced by arsenic trioxide and its correlation with Survivin gene%三氧化二砷诱导Raji细胞凋亡与Survivin基因关系的研究

    Institute of Scientific and Technical Information of China (English)

    Yi Long; Huimin Li; Chen Qing; Hua Liu; Yanli Zhang; Meijia Yu

    2008-01-01

    Objective:To investigate the apoptosis induction by arsenic trioxide (As2O3) in Raji cells and its correlation with cell cycle arrest and expression of the Survivin gene.Methods:After Raji cells were treated with As2O3 in different concentrations (1,2,4 and 8 μM),for 24,48 and 72 h,respectively,and cell proliferation was tested by MTT assay.Apoptosis was observed with electron microscope end DNA electrophoresis.The distribution of cell cycles and cell apoptosis were detected by flow cytometry.Expression of the Survivin gene was determined by real-time quantitative RT-PCR.Results:As2O3 (1-8 μM) inhibited Raji cells growth effectively in a dose- and time-dependent manner.As2O3 at 2-8μM could induce cell apoptosis and cell cycle arrest.However,As2O3 (1 μM) inhibited Raji proliferation only by cell cycle arrest,without any symptoms of cell apoptosis.At the same time,Survivin gene expression was down-regulated after the treatment.Conclusion:As2O3 could induce substantial proliferation inhibition,cell cycle arrest and apoptosis in Raji cell.Cell cycle arrest might be a reason why apoptosis occurs.As2O3 can markedly down-regulate expression of the Survivin gene in a dose- and timedependent manner.The down-regulated Survivin gene might be leading to cell apoptosis by As2O3.

  2. Survivin siRNA协同5-FU抑制MCF-7细胞的增殖%Synergism between a siRNA targeted to survivin and 5-FU in inhibiting MCF-7 cell proliferation in vitro

    Institute of Scientific and Technical Information of China (English)

    张淑群; 张淑慧; 薛兴欢; 王西京; 姜建涛

    2006-01-01

    Objective To investigate the role of small interfering RNA (siRNA) targeted to survivin in combination with 5-fluorouracil (5-FU) in inhibiting the proliferation of MCF-7 cells. Methods A siRNA targeted to survivin was synthesized and transfected into MCF-7 cells via lipofectin. Changes of the cell growth activity in response to combined treatment with survivin siRNA and 5-FU or 5-FU treatment alone was evaluated by MTT assay. The Q method of Jin Zhenjun was used to evaluated synergism between the synthesized siRNA and 5-FU. Results Treatment with 5 nmol/L siRNA reduced the IC50 of 5-FU from 4.42 to 1.18 μg/ml, and the inhibitory effect of combined treatment on MCF-7 cells was higher than that of 5-FU alone (F=26.74, P<0.01). Synergism effect (Q ≥ 1.15) was observed between 5-FU at lower concentrations and survivin siRNA. Conclusion siRNA may enhance the effectiveness of 5-FU in inhibiting the proliferation of MCF-7 cells.%目的研究以survivin为靶标的小干扰RNA(siRNA)与化疗药5-FU联合应用抑制MCF-7细胞增殖的作用.方法利用T7 RNA聚合酶体外转录合成siRNA并筛选出一条RNA干扰survivin基因的靶序列.以脂质体为载体,将survivin siRNA转染至人乳腺癌MCF-7细胞中,用四氮唑盐(MTT)法染色并计算siRNA联用5-FU对MCF-7细胞的抑制率,用SAS统计软件及金正均Q值法进行统计分析.结果单用5-FU处理细胞,其IC50为4.42 μg/ml,而加入5 nmol/L siRNA后,其IC50降为1.18 μg/ml,siRNA与5-FU联用对MCF-7细胞的抑制作用较单用5-FU明显增强(F=26.74,P<0.01);Q值分析表明survivin siRNA与中低浓度的5-FU联用,有较好的协同作用(Q≥1.15).结论survivinsiRNA可显著增强5-FU对MCF-7细胞增殖的抑制,提高肿瘤细胞对化疗药物的敏感性,克服耐药性的产生.

  3. Emmprin and survivin predict response and survival following cisplatin-containing chemotherapy in patients with advanced bladder cancer

    DEFF Research Database (Denmark)

    Als, Anne B; Dyrskjøt, Lars; von der Maase, Hans;

    2007-01-01

    in an independent material of 124 patients receiving cisplatin-containing therapy. RESULTS: Fifty-five differentially expressed genes correlated significantly to survival time. Two of the protein products (emmprin and survivin) were validated using immunohistochemistry. Multivariate analysis identified emmprin......PURPOSE: Cisplatin-containing chemotherapy is the standard of care for patients with locally advanced and metastatic transitional cell carcinoma of the urothelium. The response rate is approximately 50% and tumor-derived molecular prognostic markers are desirable for improved estimation of response...... and survival. EXPERIMENTAL DESIGN: Affymetrix GeneChip expression profiling was carried out using tumor material from 30 patients. A set of genes with an expression highly correlated to survival time after chemotherapy was identified. Two genes were selected for validation by immunohistochemistry...

  4. Emmprin and Survivin predict response and survival following cisplatin-containing chemotherapy in patients with advanced bladder cancer

    DEFF Research Database (Denmark)

    Als, Anne Birgitte; Andersen, Lars Dyrskjøt; Maase, Hans von der;

    2007-01-01

    in an independent material of 124 patients receiving cisplatin-containing therapy. RESULTS: Fifty-five differentially expressed genes correlated significantly to survival time. Two of the protein products (emmprin and survivin) were validated using immunohistochemistry. Multivariate analysis identified emmprin......PURPOSE: Cisplatin-containing chemotherapy is the standard of care for patients with locally advanced and metastatic transitional cell carcinoma of the urothelium. The response rate is approximately 50% and tumor-derived molecular prognostic markers are desirable for improved estimation of response...... and survival. EXPERIMENTAL DESIGN: Affymetrix GeneChip expression profiling was carried out using tumor material from 30 patients. A set of genes with an expression highly correlated to survival time after chemotherapy was identified. Two genes were selected for validation by immunohistochemistry...

  5. The Nuclear Staining of Survivin and Livin and Its Clinical Significance in Non-Small Cell Lung Cancer%Livin、Survivin蛋白在非小细胞肺癌胞核中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    王栋; 钱有辉; 王光锁; 任康奇

    2013-01-01

    目的 研究凋亡抑制蛋白Survivin和Livin在非小细胞肺癌(NSCLC)细胞核中阳性表达(核阳)水平及其临床意义.方法 采用免疫组织化学SP法检测40例NSCLC组织和20例癌旁正常肺组织中Survivin蛋白和Livin蛋白核阳水平,并分析其与临床特征的关系.结果 在NSCLC和癌旁正常肺组织中,Survivin蛋白细胞核表达的阳性率分别为80%(32/40)和5%(1/20),Livin蛋白细胞核表达的阳性率分别为80%(32/40)和10%(2/20).Survivin 蛋白和Livin蛋白核阳在NSCLC组织中的表达显著高于癌旁组织(P<0.05).Survivin蛋白核阳与淋巴结转移、肿瘤分期高度相关(P<0.05).结论 Survivin蛋白和Livin蛋白在NSCLC细胞核中高表达,可能成为NSCLC诊断的新指标和治疗的潜在靶点.%Objective In this study,we will investigate the nuclear staining of Survivin and livin and its clinical significance in non-small cell lung cancer (NSCLC). Methods 40 Paraffin-embedded NSCLC tissue blocks obtain randomly and 20 Paraffin-embedded bullae of lung tissue blocks were obtained for immunohistochemical staining to evaluate the expression of Survivin and Livin. The Chi-Square test was performed to evaluate the correlation of staining and clinical outcome. Results The nuclear positive rates of Survivin in NSCLC and bullae of lung tissue were 80% (32/40) and 5% (1/20), respectively. The nuclear positive rates of Livin in NSCLC and bullae of lung tissue were 80% (32/40) and 10% (2/20), respectively. The nuclear staining of Survivin and Livin were higher in NSCLC tissue samples than that in bullae of lung samples (P<0.05). The nuclear staining of Survivin was associate with lymph node metastasis and stage of tumor (P<0.05). Conclusions The expressions of Survivin and Livin were high in NSCLC. It will provide a new target for the diagnosis and treatment of lung carcinoma.

  6. Rapamycin potentiates cytotoxicity by docetaxel possibly through downregulation of Survivin in lung cancer cells

    Directory of Open Access Journals (Sweden)

    Li Hui

    2011-03-01

    Full Text Available Abstract Background To elucidate whether rapamycin, the inhibitor of mTOR (mammalian target of rapamycin, can potentiate the cytotoxic effect of docetaxel in lung cancer cells and to probe the mechanism underlying such enhancement. Methods Lung cancer cells were treated with docetaxel and rapamycin. The effect on the proliferation of lung cancer cells was evaluated using the MTT method, and cell apoptosis was measured by flow cytometry. Protein expression and level of phosphorylation were assayed using Western Blot method. Results Co-treatment of rapamycin and docetaxel was found to favorably enhance the cytotoxic effect of docetaxel in four lung cancer cell lines. This tumoricidal boost is associated with a reduction in the expression and phosphorylation levels of Survivin and ERK1/2, respectively. Conclusion The combined application of mTOR inhibitor and docetaxel led to a greater degree of cancer cell killing than that by either compound used alone. Therefore, this combination warrants further investigation in its suitability of serving as a novel therapeutic scheme for treating advanced and recurrent lung cancer patients.

  7. Specific survivin dual fluorescence resonance energy transfer molecular beacons for detection of human bladder cancer cells

    Institute of Scientific and Technical Information of China (English)

    Zhi-qiang WANG; Jun ZHAO; Jin ZENG; Kai-jie WU; Yu-le CHEN; Xin-ya ng WANG; Luke S CHANG; Da-lin HE

    2011-01-01

    Survivin molecular beacons can be used to detectbladder cancer cells in urine samples non-invasively.The aim of this study is to improve the specificity of detection of bladder cancer cells using survivin dual fluorescence resonance energy transfer molecular beacons (FRET MBs) that have fluorophores forming one donor-acceptor pair.Methods:Survivin-targeting dual fluorescence resonance energy transfer molecular beacons with unique target sequences were designed,which had no overlap with the other genes in the apoptosis inhibitor protein family.Human bladder cancer cell lines 5637,253J and T24,as well as the exfoliated cells in the urine of healthy adults and patients with bladder cancer were examined.Images of cells were taken using a laser scanning confocal fluorescence microscope.For assays using dual FRET MBs,the excitation wavelength was 488 nm,and the emission detection wavelengths were 520+20 nm and 560+20 nm,respectively.Results:The human bladder cancer cell lines and exfoliated cells in the urine of patients with bladder cancer incubated with the survivin dual FRET MBs exhibited strong fluorescence signals.In contrast,no fluorescence was detected in the survivin-negative human dermal fibroblasts-adult (HDF-a) cells or exfoliated cells in the urine of healthy adults incubated with the survivin dual FRET MBs.Conclusion:The results suggest that the survivin dual FRET MBs may be used as a specific and non-invasive method for early detection and follow-up of patients with bladder cancer.

  8. Downregulation of Epidermal Growth Factor Receptor Expression Contributes to α-TEA's Proapoptotic Effects in Human Ovarian Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Ming-Chieh Shun

    2010-01-01

    Full Text Available RRR-α-tocopherol derivative α-TEA (RRR-α-tocopherol ether-linked acetic acid analog has been shown to be a potent antitumor agent both in vivo and in vitro. In this study, we investigated the effects of α-TEA on the expression of epidermal growth factor receptor (EGFR family members, ErbB1, 2 and 3, and the role of ErbB 2 and 3 in α-TEA-induced apoptosis and suppression of Akt, FLIP and survivin in the cisplatin-sensitive (A2780S and -resistant (A2780/CP70R human ovarian cancer cell lines. Data show that α-TEA's ability to induced apoptosis was associated with reduced expression of ErbB1 (cisplatin-resistant cells, 2 and 3 (both cell types and reduced levels of the phosphorylated (active form of Akt; as well as, reduced levels of FLIP and survivin proteins in both cell types. Ectopic overexpression and siRNA knockdown studies showed that ErbB2, ErbB3, Akt, FLIP and survivin are involved in α-TEA-induce apoptosis and that α-TEA downregulates FLIP and survivin via suppression of pAkt, which is mediated by ErbB2 and ErB3. Thus, α-TEA is a potent pro-apoptotic agent for both cisplatin-sensitive and -resistant ovarian cancer cell lines in cell culture and it produces cell death, at least in part, by downregulation of members of the EGFR family.

  9. Regulation of Aicda expression and AID activity.

    Science.gov (United States)

    Zan, Hong; Casali, Paolo

    2013-03-01

    Activation-induced cytidine deaminase (AID) is expressed in a B cell differentiation stage-specific fashion and is essential for immunoglobulin (Ig) gene class switch DNA recombination (CSR) and somatic hypermutation (SHM). CSR and SHM play a central role in the maturation of antibody and autoantibody responses. AID displays a mutagenic activity by catalyzing targeted deamination of deoxycytidine (dC) residues in DNA resulting in dU:dG mismatches, which are processed into point-mutations in SHM or double-strand breaks (DSBs) in CSR. Although AID specifically targets the Ig gene loci (IgH, Igκ and Igλ), it can also home into a wide array of non-Ig genes in B-and non-B-cell backgrounds. Aberrant expression of AID is associated with multiple diseases such as allergy, inflammation, autoimmunity and cancer. In autoimmune systemic lupus erythematosus, dysregulated AID expression underpins increased CSR, SHM and autoantibody production. As a potent mutator, AID is under stringent transcriptional, post-transcriptional and post-translational regulation. AID is also regulated in its targeting and enzymatic function. In resting naïve or memory B cells, AID transcripts and protein are undetectable. These, however, are readily and significantly up-regulated in B cells induced to undergo CSR and/or SHM. Transcription factors, such as HoxC4 and NF-κB, which are up-regulated in a B cell lineage-and/or differentiation stage-specific manner, regulate the induction of AID. HoxC4 induces AID expression by directly binding to the AID gene promoter through an evolutionarily conserved 5'-ATTT-3' motif. HoxC4 is induced by the same stimuli that induce AID and CSR. It is further up-regulated by estrogen through three estrogen responsive elements in its promoter region. The targeting of AID to switch (S) regions is mediated by 14-3-3 adaptor proteins, which specifically bind to 5'-AGCT-3' repeats that are exist at high frequency in S region cores. Like HoxC4, 14-3-3 adaptors are induced

  10. 慢病毒介导TIEG1基因乳腺癌靶向载体的构建及活性测定%Construction of lentiviral vector expressing TIEG1 gene with breast cancer specific promoter and identification of activity

    Institute of Scientific and Technical Information of China (English)

    蒋磊; 林飞燕; 王福乐; 叶爱芳; 吴建波

    2011-01-01

    Objective Construction of lentivral vector expressing TIEG1 gene with survivin promoter and identifition of its activity. Methods Survivin promoter and TIEG1 gene full length fragment were obtained by PCR amplification. Lentivial vector containing TIEG1 gene and survivin promoter were constructed. The inserted fragment was verified by double enzyme digestion and DNA sequencing. The VSVG pseudotyped lentiviruses were produced and transduced into SK-BR-3 breast cancer cells and normal HUVEC cells. ECJFP expression was examined by fluorescence microscopy. Results The lentiviral TIEG1 gene expression vector with survivin promoter was constructed successfully and transduced into SK-BR-3 and HUVEC cells. Green fluorescence was observed in SK-BR-3 cells and not in HUVEC cells. Overexpression of TIEG1 gene was verified by RT-PCR in SK-BR-3 cells. Conclusions These data suggest that the survivin promoter drive the specific gene expression in breast cancer cell, which are promising candidates in targeted tumor gene therapy.%目的:构建慢病毒介导的TIEG1基因乳腺癌特异性靶向载体,并鉴定其活性.方法:将乳腺癌特异性survivin启动子片段和TIEG1基因先后克隆进带有绿色荧光蛋白的慢病毒载体中,构建survivin启动子驱动的TIEG1基因慢病毒表达载体,酶切及测序鉴定.包装纯化慢病毒颗粒,感染人脐静脉内皮细胞HUVEC和乳腺癌细胞SK-BR-3,观察绿色荧光蛋白的特异性表达.结果:成功构建带有肿瘤特异性survivin启动子驱动的TIEG1基因慢病毒表达载体.感染细胞后,乳腺癌SK-BR-3细胞可观察到绿色荧光表达,而人正常脐静脉内皮细胞基本无表达.半定量RT-PCR证实TIEG1基因在乳腺癌SK-BR-3细胞中过表达.结论:构建的survivin启动子驱动的慢病毒表达载体具有一定的肿瘤特异性,将为实现以慢病毒为载体的TIEG1基因肿瘤靶向治疗提供良好的实验基础.

  11. Active inflammatory biomarkers in oral lichen planus.

    Science.gov (United States)

    Santarelli, Andrea; Mascitti, Marco; Rubini, Corrado; Bambini, Fabrizio; Zizzi, Antonio; Offidani, Annamaria; Ganzetti, Giulia; Laino, Luigi; Cicciù, Marco; Lo Muzio, Lorenzo

    2015-12-01

    Oral lichen planus (OLP) is a chronic disease, with a central role to cell-mediated autoimmunity. Osteopontin promotes migration and recruitment of immune cells, CD44 is its receptor, and Survivin seems to be important in skin/mucosa homeostasis. The aim of this study was to investigate their expression in biopsy specimens of patients with different OLP clinical types and healthy controls.Biopsy specimens from 30 patients with OLP (15 atrophic and 15 hyperplastic) and 15 healthy controls were subjected to immune-histochemical analysis, to detect the expression of osteopontin, CD44, and Survivin in oral epithelia. The distributions of positively stained cells were evaluated with a quantitative method, while the inflammation degree was evaluated with a semi-quantitative one.Expression of osteopontin and CD44 was higher in OLP than controls, while Survivin expression was lower in OLP patients. There was a greater reduction of Survivin expression in atrophic OLP than hyperplastic OLP. A correlation between osteopontin expression and a high degree of inflammation was found. Furthermore, Survivin expression was higher in cases with low intensity of inflammation.Osteopontin, CD44, and Survivin seem to be involved in OLP pathogenesis, and further investigations are needed for clarifying their role in this oral disease.

  12. Tension sensing by Aurora B kinase is independent of survivin-based centromere localization.

    Science.gov (United States)

    Campbell, Christopher S; Desai, Arshad

    2013-05-01

    Accurate segregation of the replicated genome requires chromosome biorientation on the spindle. Biorientation is ensured by Aurora B kinase (Ipl1), a member of the four-subunit chromosomal passenger complex (CPC). Localization of the CPC to the inner centromere is central to the current model for how tension ensures chromosome biorientation: kinetochore-spindle attachments that are not under tension remain close to the inner centromere and are destabilized by Aurora B phosphorylation, whereas kinetochores under tension are pulled away from the influence of Aurora B, stabilizing their microtubule attachments. Here we show that an engineered truncation of the Sli15 (known as INCENP in humans) subunit of budding yeast CPC that eliminates association with the inner centromere nevertheless supports proper chromosome segregation during both mitosis and meiosis. Truncated Sli15 suppresses the deletion phenotypes of the inner-centromere-targeting proteins survivin (Bir1), borealin (Nbl1), Bub1 and Sgo1 (ref. 6). Unlike wild-type Sli15, truncated Sli15 localizes to pre-anaphase spindle microtubules. Premature targeting of full-length Sli15 to microtubules by preventing Cdk1 (also known as Cdc28) phosphorylation also suppresses the inviability of Bir1 deletion. These results suggest that activation of Aurora B kinase by clustering either on chromatin or on microtubules is sufficient for chromosome biorientation.

  13. The impact of SiRNA-induced silencing of CD147 on the expression of survivin in HepG2 cells%SiRNA抑制肝癌细胞株HepG2中CD147基因表达对survivin表达的影响

    Institute of Scientific and Technical Information of China (English)

    陈志刚; 周雄心

    2009-01-01

    目的:探讨CD147与survivin(SVV)的关系.方法:设计、合成两对CD147编码基因的反向重复序列,运用瞬时转染方法抑制HepG2中CD147表达,运用RT-PCR、Western blot方法检测干扰后CD147、SVV及caspase-3表达的改变,流式细胞术检测干扰后肿瘤细胞的凋亡情况.结果:SiRNA sequence 1、2均可有效地抑制CD147基因的表达(P<0.05),伴随着SVV mRNA和蛋白表达水平降低(P<0.05);干扰后caspase-3 mRNA水平升高(P<0.05),活性caspase-3蛋白水平增高(P<0.05),肿瘤细胞凋亡增加,以细胞早期凋亡改变最为明显(P<0.05).结论:沉默HepG2中CD147基因能下调SVV基因表达,致使肿瘤细胞凋亡增加.CD147与SVV在细胞内可能存在着相互调节机制,然而其确切机制还有待进一步研究.

  14. Livin基因和Survivin基因联合靶向siRNA抑制大肠癌细胞增殖的作用%Inhibitory effect of multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes on human: colon cancer cells proliferation

    Institute of Scientific and Technical Information of China (English)

    蔡明; 王国斌; 陶凯雄; 蔡昌学

    2009-01-01

    采用1个载体编码2个shRNA技术,构建Livin基因和Survivin基因联合靶向的siRNA重组表达载体,研究其对人大肠癌细胞生长协同抑制的增效作用.构建Livin基因和Survivin基凶联合靶向的siRNA重组表达载体并转染大肠癌细胞,通过RT-PCR和Western blotting方法检测Livin基因与Survivin基因的表达,利用流式细胞仪榆测处理后细胞的凋亡效应.经酶切鉴定和测序分析证实Livin基因和Survivin基因联介靶向的siRNA重组表达载体构建成功,它对大肠癌细胞Livin和Survivin mRNA的抑制率分别为27.90%和32.24%,对Livin和Survivin蛋白表达的抑制率分别为22.28%和40.86%,诱导肿瘤细胞凋亡率为(11.69±1.37)%,但协同十扰作用比单独干扰Livin 基因或Survivin基因弱.Livin基因和Survivin基冈联合靶向的siRNA重组表达载体构建成功并能抑制Livin基因和Survivin基因的表达,但协同抑制作用比单独干扰Livin基因或Survivin基因弱.

  15. Survivin and NAIP in Human Benign Prostatic Hyperplasia: Protective Role of the Association of Serenoa repens, Lycopene and Selenium from the Randomized Clinical Study

    Science.gov (United States)

    Morgia, Giuseppe; Micali, Antonio; Rinaldi, Mariagrazia; Irrera, Natasha; Marini, Herbert; Puzzolo, Domenico; Pisani, Antonina; Privitera, Salvatore; Russo, Giorgio I.; Cimino, Sebastiano; Ieni, Antonio; Trichilo, Vincenzo; Altavilla, Domenica; Squadrito, Francesco; Minutoli, Letteria

    2017-01-01

    Benign prostatic hyperplasia (BPH) treatment includes the apoptosis machinery modulation through the direct inhibition of caspase cascade. We previously demonstrated that Serenoa repens (Ser) with lycopene (Ly) and selenium (Se) reawakened apoptosis by reducing survivin and neuronal apoptosis inhibitory protein (NAIP) levels in rats. The aim of this study was to evaluate the effectiveness of Ser-Se-Ly association on survivin and NAIP expression in BPH patients. Ninety patients with lower urinary tract symptoms (LUTS) due to clinical BPH were included in this randomized, double-blind, placebo-controlled trial. Participants were randomly assigned to receive placebo (Group BPH + placebo, n = 45) or Ser-Se-Ly association (Group BPH + Ser-Se-Ly; n = 45) for 3 months. At time 0, all patients underwent prostatic biopsies. After 3 months of treatment, they underwent prostatic re-biopsy and specimens were collected for molecular, morphological, and immunohistochemical analysis. After 3 months, survivin and NAIP were significantly decreased, while caspase-3 was significantly increased in BPH patients treated with Ser-Se-Ly when compared with the other group. In BPH patients treated with Ser-Se-Ly for 3 months, the glandular epithelium was formed by a single layer of cuboidal cells. PSA showed high immunoexpression in all BPH patients and a focal positivity in Ser-Se-Ly treated patients after 3 months. Evident prostate specific membrane antigen (PSMA) immunoexpression was shown in all BPH patients, while no positivity was present after Ser-Se-Ly administration. Ser-Se-Ly proved to be effective in promoting apoptosis in BPH patients. PMID:28327526

  16. Survivin and NAIP in Human Benign Prostatic Hyperplasia: Protective Role of the Association of Serenoa repens, Lycopene and Selenium from the Randomized Clinical Study

    Directory of Open Access Journals (Sweden)

    Giuseppe Morgia

    2017-03-01

    Full Text Available Benign prostatic hyperplasia (BPH treatment includes the apoptosis machinery modulation through the direct inhibition of caspase cascade. We previously demonstrated that Serenoa repens (Ser with lycopene (Ly and selenium (Se reawakened apoptosis by reducing survivin and neuronal apoptosis inhibitory protein (NAIP levels in rats. The aim of this study was to evaluate the effectiveness of Ser-Se-Ly association on survivin and NAIP expression in BPH patients. Ninety patients with lower urinary tract symptoms (LUTS due to clinical BPH were included in this randomized, double-blind, placebo-controlled trial. Participants were randomly assigned to receive placebo (Group BPH + placebo, n = 45 or Ser-Se-Ly association (Group BPH + Ser-Se-Ly; n = 45 for 3 months. At time 0, all patients underwent prostatic biopsies. After 3 months of treatment, they underwent prostatic re-biopsy and specimens were collected for molecular, morphological, and immunohistochemical analysis. After 3 months, survivin and NAIP were significantly decreased, while caspase-3 was significantly increased in BPH patients treated with Ser-Se-Ly when compared with the other group. In BPH patients treated with Ser-Se-Ly for 3 months, the glandular epithelium was formed by a single layer of cuboidal cells. PSA showed high immunoexpression in all BPH patients and a focal positivity in Ser-Se-Ly treated patients after 3 months. Evident prostate specific membrane antigen (PSMA immunoexpression was shown in all BPH patients, while no positivity was present after Ser-Se-Ly administration. Ser-Se-Ly proved to be effective in promoting apoptosis in BPH patients.

  17. Performance of survivin mRNA as a biomarker for bladder cancer in the prospective study UroScreen.

    Directory of Open Access Journals (Sweden)

    Georg Johnen

    Full Text Available BACKGROUND: Urinary biomarkers have the potential to improve the early detection of bladder cancer. Most of the various known markers, however, have only been evaluated in studies with cross-sectional design. For proper validation a longitudinal design would be preferable. We used the prospective study UroScreen to evaluate survivin, a potential biomarker that has multiple functions in carcinogenesis. METHODS/RESULTS: Survivin was analyzed in 5,716 urine samples from 1,540 chemical workers previously exposed to aromatic amines. The workers participated in a surveillance program with yearly examinations between 2003 and 2010. RNA was extracted from urinary cells and survivin was determined by Real-Time PCR. During the study, 19 bladder tumors were detected. Multivariate generalized estimation equation (GEE models showed that β-actin, representing RNA yield and quality, had the strongest influence on survivin positivity. Inflammation, hematuria and smoking did not confound the results. Survivin had a sensitivity of 21.1% for all and 36.4% for high-grade tumors. Specificity was 97.5%, the positive predictive value (PPV 9.5%, and the negative predictive value (NPV 99.0%. CONCLUSIONS: In this prospective and so far largest study on survivin, the marker showed a good NPV and specificity but a low PPV and sensitivity. This was partly due to the low number of cases, which limits the validity of the results. Compliance, urine quality, problems with the assay, and mRNA stability influenced the performance of survivin. However, most issues could be addressed with a more reliable assay in the future. One important finding is that survivin was not influenced by confounders like inflammation and exhibited a relatively low number of false-positives. Therefore, despite the low sensitivity, survivin may still be considered as a component of a multimarker panel.

  18. Competitive inhibition of survivin using a cell-permeable recombinant protein induces cancer-specific apoptosis in colon cancer model

    Directory of Open Access Journals (Sweden)

    Roy K

    2015-02-01

    Full Text Available Kislay Roy,1 Rupinder K Kanwar,1 Subramanian Krishnakumar,2,3 Chun Hei Antonio Cheung,4 Jagat R Kanwar1 1Nanomedicine-Laboratory of Immunology and Molecular Biomedical Research (NLIMBR, Molecular and Medical Research (MMR Strategic Research Centre, School of Medicine (SoM, Faculty of Health, Deakin University, Waurn Ponds, VIC, Australia; 2Department of Nanobiotechnology, 3Larsen & Toubro (L&T Ocular Pathology Department, Vision Research Foundation, Kamalnayan Bajaj Institute for Research in Vision and Ophthalmology, Chennai, India; 4Department of Pharmacology, College of Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China Abstract: Endogenous survivin expression has been related with cancer survival, drug resistance, and metastasis. Therapies targeting survivin have been shown to significantly inhibit tumor growth and recurrence. We found out that a cell-permeable dominant negative survivin (SurR9-C84A, referred to as SR9 competitively inhibited endogenous survivin and blocked the cell cycle at the G1/S phase. Nanoencapsulation in mucoadhesive chitosan nanoparticles (CHNP substantially increased the bioavailability and serum stability of SR9. The mechanism of nanoparticle uptake was studied extensively in vitro and in ex vivo models. Our results confirmed that CHNP–SR9 protected primary cells from autophagy and successfully induced tumor-specific apoptosis via both extrinsic and intrinsic apoptotic pathways. CHNP–SR9 significantly reduced the tumor spheroid size (three-dimensional model by nearly 7-fold. Effects of SR9 and CHNP–SR9 were studied on 35 key molecules involved in the apoptotic pathway. Highly significant (4.26-fold, P≤0.005 reduction in tumor volume was observed using an in vivo mouse xenograft colon cancer model. It was also observed that net apoptotic (6.25-fold, P≤0.005 and necrotic indexes (3.5-fold, P≤0.05 were comparatively higher in CHNP–SR9 when compared to void CHNP and CHNP–SR9

  19. Expression of survivin and SIRT1 mRNA and their correlation in non-small cell lung cancer%Survivin和SIRT1 mRNA在非小细胞肺癌中的表达及两者相关性分析

    Institute of Scientific and Technical Information of China (English)

    毛旭华; 汤俊明; 井昶雯; 曹海霞

    2016-01-01

    目的 检测survivin和SIRT1 mRNA在非小细胞肺癌中的表达,探讨其相互关系.方法 用实时定量PCR的方法检测江苏省肿瘤医院42组非小细胞肺癌组织和对应的癌旁组织中survivin和SIRT1 mRNA的表达,统计survivin与SIRT1的相关性.观察EGFR酪氨酸酶抑制剂吉非替尼处理非小细胞肺癌细胞系HCC827后survivin和SIRT1的表达改变.结果 85.7% (36/42)肿瘤组织survivin mRNA表达明显高于对应的癌旁组织.61.9% (26/42)肿瘤组织SIRT1 mRNA的表达低于其癌旁组织,33.3%(14/42)肿瘤组织SIRT1 mRNA表达高于其癌旁组织.Survivin表达与SIRT1表达有统计学意义的相关性(P<0.05).吉非替尼能增加HCC827细胞中的SIRT1表达,降低survivin的表达水平,呈时间和剂量依赖性.结论 Survivin在非小细胞肺癌组织中高表达,而大部分SIRT1基因呈低表达状态.survivin与SIRT1表达呈负相关.在细胞水平上,吉非替尼能增加SIRT1的表达从而降低survivin的水平.提示两者在非小细胞肺癌的发生发展中可能发挥重要作用.

  20. Survivin mRNA antagonists using locked nucleic acid, potential for molecular cancer therapy

    DEFF Research Database (Denmark)

    Fisker, Niels; Westergaard, Majken; Hansen, Henrik Frydenlund;

    2007-01-01

    synergistic effect when combining the mRNA antagonists against Survivin with the chemotherapeutic Taxol. This effect was demonstrated at concentrations of antagonists far lower than any previously demonstrated, indicating the high potential of locked nucleic acid for therapeutic use. Further characterisations...

  1. Survivin mediates targeting of the chromosomal passenger complex to the centromere and midbody

    NARCIS (Netherlands)

    Vader, G; Kauw, JJW; Medema, RH; Lens, SMA

    2006-01-01

    The chromosomal passenger complex (CPC) coordinates chromosomal and cytoskeletal events of mitosis. The enzymatic core of this complex (Aurora-B) is guided through the mitotic cell by its companion chromosomal passenger proteins, inner centromere protein (INCENP), Survivin and Borealin/Dasra-B, ther

  2. Role of Survivin associated with clinicopathological features in cervical cancer:a Meta-analysis%Survivin蛋白和宫颈癌风险荟萃分析

    Institute of Scientific and Technical Information of China (English)

    王燕琴; 王志莲; 郝敏

    2016-01-01

    ObjectiveTo evaluate the relationship between expression of Survivin and cervical cancer.MethodsUsed the keywords "surviving", "cervical cancer", "cervical carcinoma", "cancer of cervix" "Survivin", "cervical cancer", "cancer of the cervix", to search Pubmed, Embase, CNKI (China National Knowledge Infrastructure) for studies up to September 2015.ResultsSixteen articles were included, including 1 071 patients, of which 780 cervical cancer cases and 291 normal control cases. The results wereRRs (Risk Ratios)=0.215, and 95%CI (confidence interval) 0.100-0.459,P<0.001. In subgroup analysis, IHC (immunihistochemistry) GroupP=0.001, ISH (in situ hybridization) Group P=0.038, were statistically significant. And RT-PCR (Real time-Polymerase Chain Reaction) +IHC group wasP=0.300, the difference was not statistically significant. Survivin protein location subgroup,I2=75.7%, Cyto and Nu/Cyto group wasI2=74.8%, no-reported group wasI2=67.8%, suggesting that protein location existed heterogeneity. After eliminating three heterogeneity of studies, the pooled data wereRR=0.15, 95%CI 0.01-1.57, theP-value did not changed. The result was stable.ConclusionsSurvivin expression was significantly correlated with cervical cancer, which played an important role in the development of cervical cancer. Survivin could be targeted as a treatment for cervical cancer.%目的:评估Survivin蛋白和宫颈癌发病风险的相关性。方法利用关键词“Survivin、存活蛋白、子宫颈癌”,英文检索词包括“uterine cervical neoplasms、cervical cancer、Survivin”,通过Pubmed、Embase、CNKI(China National Knowledge Infrastructure)进行文献检索。检索时限至2015年9月15日。结果最终纳入16篇文献,1071例患者,其中宫颈癌780例、正常对照291例。宫颈癌和正常对照Survivin蛋白表达差异有统计学意义,RR=0.21、95%CI 0.10~0.46,P=0.000。亚组分析,免疫组化组(IHC 组)P=0.001,原位杂交组(ISH 组)P=0

  3. Pro-apoptosis Effect of Survivin T34A Mutant on Cancer Cells in Vitro and Vivo

    Institute of Scientific and Technical Information of China (English)

    SUN Jing; MA Lan; WU Hao; LIU Lian-ke

    2014-01-01

    Objective:To explore the pro-apoptosis effect of Survivin T34A mutant on cancer cells in vitro and vivo. Methods: After highly-metastasized breast cancer cells (4T1 cells) were transfected with Survivin T34A plasmid and wild survivin plasmid, cell proliferation was analyzed using tetrazolium blue (MTT) assay and apoptosis was detected with lfow cytometry. In animal experiments, mice were vaccinated subcutaneously with 4T1 cells and treated with T34A plasmid and wild survivin plasmid. The tumor volume and weight, wet weight of the lung, number of pulmonary metastasis nodule were measured. H&E staining and TUNEL detection of tumor apoptosis were performed after mice were executed. Results: The cell survival rate was signiifcantly decreased (P<0.01) and apoptotic rate increased (P<0.01) after treatment with Survivin T34A plasmid in vitro. In vivo, 4 days after treatment, tumor volume was signiifcantly smaller, mean tumor weight and mean wet weight of the lung were obviously lighter, and pulmonary metastasis nodule was evidently fewer (P<0.05). The apoptotic cells and large areas of necrosis were observed, and apoptotic index was increased (P<0.05). Conclusion:Survivin T34A can induce the apoptosis of 4T1 cells with specificity and may become a new approach to breast cancer.

  4. Robust, synergistic regulation of human gene expression using TALE activators.

    Science.gov (United States)

    Maeder, Morgan L; Linder, Samantha J; Reyon, Deepak; Angstman, James F; Fu, Yanfang; Sander, Jeffry D; Joung, J Keith

    2013-03-01

    Artificial activators designed using transcription activator-like effector (TALE) technology have broad utility, but previous studies suggest that these monomeric proteins often exhibit low activities. Here we demonstrate that TALE activators can robustly function individually or in synergistic combinations to increase expression of endogenous human genes over wide dynamic ranges. These findings will encourage applications of TALE activators for research and therapy, and guide design of monomeric TALE-based fusion proteins.

  5. Study on effect of survivin antioligonucleotide on subcutaneous transplanted Xunwei lung adenocarcinoma nude mice model%Survivin反义核苷酸对宣威肺腺癌移植瘤裸鼠模型作用的研究

    Institute of Scientific and Technical Information of China (English)

    王巍炜; 洪志鹏; 李高峰; 王绍佳; 陈瑞彬; 张继朋

    2012-01-01

    目的 选用宣威肺腺癌细胞裸鼠移植瘤模型,采用survivinmRNA的反义寡核苷酸(ASODN)抑制宣威肺腺癌细胞,观察其疗效,为进一步临床研究提供实验依据.方法 建立宣威肺腺癌细胞XWLC-05裸鼠皮下移植瘤模型,选取48只成瘤裸鼠,随机分为:对照组、单纯脂质体转染(LIP)组、survivin正义寡核苷酸(SODN)组、survivinASODN组,通过皮下移植瘤内多点注射后,观察裸鼠一般情况、移植瘤体积,计算抑瘤率等.病理学检测各组肿瘤的变化以及对心脏、肝脏、肾脏的影响.结果 survivinASODN组注射survivinASODN后,裸小鼠表现为生长速度较为缓慢,质量低于其他3组(P>0.05),其生长抑制率高于其他3组(P>0.05).而对照组、LIP组、survivinSODN组在肿瘤质量及生长抑制率方面均无明显差异.病理检查见瘤组织中有大量的细胞坏死灶.各组裸鼠均未出现死亡,病理检查重要脏器未见明显损害.结论 survivinASODN能够抑制裸鼠皮下移植瘤的生长,未对其他脏器产生明显损害.%Objective By selecting transplanted Xunwei lung adenocarcinoma nude mice model, to adopt survivinnmRNA anti-sense oligonucleotide to inhibit Xunwei lung adenocarcinoma cells and to observe its effects to provide the experiment evidence for further clinical study be offered in clinical treatment. Methods After modeling subcutaneous transplanted Xunwei lung adenocarcinoma cell XWLC-05 nude mice,48 nude mice were selected and devided into 4 groups randomly: control group (blank) , Lip group (simple liposome) , survivin-SODN group (injecting sense oligonucleotide) and survivin-ASODN group (injecting antisense oligonucleotide). We injected different drugs according to different groups. Then the general activity of nude mice in different groups was observed and the tumor size was measured. Also,the antineoplastic rates in different groups were calculated. The pathological methods were used to analyse the effects of 4

  6. Heparanase expression upregulates platelet adhesion activity and thrombogenicity

    Science.gov (United States)

    Österholm, Cecilia; Zhang, Xiao; Hedin, Ulf; Vlodavsky, Israel; Li, Jin-Ping

    2016-01-01

    Heparanase is an endo-glucuronidase that specifically cleaves heparan sulfate (HS) and heparin polysaccharides. The enzyme is expressed at low levels in normal tissues, but is often upregulated under pathological conditions such as cancer and inflammation. Normal human platelets express exceptionally high levels of heparanase, but the functional consequences of this feature remain unknown. We investigated functional roles of heparanase by comparing the properties of platelets expressing high (Hpa-tg) or low (Ctr) levels of heparanase. Upon activation, Hpa-tg platelets exhibited a much stronger adhesion activity as compared to Ctr platelets, likely contributing to a higher thrombotic activity in a carotid thrombosis model. Furthermore, we found concomitant upregulated expression of both heparanase and CD62P (P-selectin) upon activation of mouse and human platelets. As platelets play important roles in tumor metastasis, these findings indicate contribution of the platelet heparanase to hyper-thrombotic conditions often seen in patients with metastatic cancer. PMID:27129145

  7. Evaluation of alpha 1-antitrypsin and the levels of mRNA expression of matrix metalloproteinase 7, urokinase type plasminogen activator receptor and COX-2 for the diagnosis of colorectal cancer.

    Directory of Open Access Journals (Sweden)

    Luis Bujanda

    Full Text Available BACKGROUND: Colorectal cancer (CRC is the second most common cause of death from cancer in both men and women in the majority of developed countries. Molecular tests of blood could potentially provide this ideal screening tool. AIM: Our objective was to assess the usefulness of serum markers and mRNA expression levels in the diagnosis of CRC. METHODS: In a prospective study, we measured mRNA expression levels of 13 markers (carbonic anhydrase, guanylyl cyclase C, plasminogen activator inhibitor, matrix metalloproteinase 7 (MMP7, urokinase-type plasminogen activator receptor (uPAR, urokinase-type plasminogen activator, survivin, tetranectin, vascular endothelial growth factor (VEGF, cytokeratin 20, thymidylate synthase, cyclooxygenase 2 (COX-2, and CD44 and three proteins in serum (alpha 1 antitrypsin, carcinoembryonic antigen (CEA and activated C3 in 42 patients with CRC and 33 with normal colonoscopy results. RESULTS: Alpha 1-antitrypsin was the serum marker that was most useful for CRC diagnosis (1.79 ± 0.25 in the CRC group vs 1.27 ± 0.25 in the control group, P<0.0005. The area under the ROC curve for alpha 1-antitrypsin was 0.88 (0.79-0.96. The mRNA expression levels of five markers were statistically different between CRC cases and controls: those for which the ROC area was over 75% were MMP7 (0.81 and tetranectin (0.80, COX-2 (0.78, uPAR (0.78 and carbonic anhydrase (0.77. The markers which identified early stage CRC (Stages I and II were alpha 1-antitrypsin, uPAR, COX-2 and MMP7. CONCLUSIONS: Serum alpha 1-antitrypsin and the levels of mRNA expression of MMP7, COX-2 and uPAR have good diagnostic accuracy for CRC, even in the early stages.

  8. Neuronal Activity Regulates Hippocampal miRNA Expression

    Science.gov (United States)

    Eacker, Stephen M.; Keuss, Matthew J.; Berezikov, Eugene; Dawson, Valina L.; Dawson, Ted M.

    2011-01-01

    Neuronal activity regulates a broad range of processes in the hippocampus, including the precise regulation of translation. Disruptions in proper translational control in the nervous system are associated with a variety of disorders that fall in the autistic spectrum. MicroRNA (miRNA) represent a relatively recently discovered player in the regulation of translation in the nervous system. We have conducted an in depth analysis of how neuronal activity regulates miRNA expression in the hippocampus. Using deep sequencing we exhaustively identify all miRNAs, including 15 novel miRNAs, expressed in hippocampus of the adult mouse. We identified 119 miRNAs documented in miRBase but less than half of these miRNA were expressed at a level greater than 0.1% of total miRNA. Expression profiling following induction of neuronal activity by electroconvulsive shock demonstrates that most miRNA show a biphasic pattern of expression: rapid induction of specific mature miRNA expression followed by a decline in expression. These results have important implications into how miRNAs influence activity-dependent translational control. PMID:21984899

  9. Neuronal activity regulates hippocampal miRNA expression.

    Directory of Open Access Journals (Sweden)

    Stephen M Eacker

    Full Text Available Neuronal activity regulates a broad range of processes in the hippocampus, including the precise regulation of translation. Disruptions in proper translational control in the nervous system are associated with a variety of disorders that fall in the autistic spectrum. MicroRNA (miRNA represent a relatively recently discovered player in the regulation of translation in the nervous system. We have conducted an in depth analysis of how neuronal activity regulates miRNA expression in the hippocampus. Using deep sequencing we exhaustively identify all miRNAs, including 15 novel miRNAs, expressed in hippocampus of the adult mouse. We identified 119 miRNAs documented in miRBase but less than half of these miRNA were expressed at a level greater than 0.1% of total miRNA. Expression profiling following induction of neuronal activity by electroconvulsive shock demonstrates that most miRNA show a biphasic pattern of expression: rapid induction of specific mature miRNA expression followed by a decline in expression. These results have important implications into how miRNAs influence activity-dependent translational control.

  10. Active AU Based Patch Weighting for Facial Expression Recognition

    Science.gov (United States)

    Xie, Weicheng; Shen, Linlin; Yang, Meng; Lai, Zhihui

    2017-01-01

    Facial expression has many applications in human-computer interaction. Although feature extraction and selection have been well studied, the specificity of each expression variation is not fully explored in state-of-the-art works. In this work, the problem of multiclass expression recognition is converted into triplet-wise expression recognition. For each expression triplet, a new feature optimization model based on action unit (AU) weighting and patch weight optimization is proposed to represent the specificity of the expression triplet. The sparse representation-based approach is then proposed to detect the active AUs of the testing sample for better generalization. The algorithm achieved competitive accuracies of 89.67% and 94.09% for the Jaffe and Cohn–Kanade (CK+) databases, respectively. Better cross-database performance has also been observed. PMID:28146094

  11. Active AU Based Patch Weighting for Facial Expression Recognition

    Directory of Open Access Journals (Sweden)

    Weicheng Xie

    2017-01-01

    Full Text Available Facial expression has many applications in human-computer interaction. Although feature extraction and selection have been well studied, the specificity of each expression variation is not fully explored in state-of-the-art works. In this work, the problem of multiclass expression recognition is converted into triplet-wise expression recognition. For each expression triplet, a new feature optimization model based on action unit (AU weighting and patch weight optimization is proposed to represent the specificity of the expression triplet. The sparse representation-based approach is then proposed to detect the active AUs of the testing sample for better generalization. The algorithm achieved competitive accuracies of 89.67% and 94.09% for the Jaffe and Cohn–Kanade (CK+ databases, respectively. Better cross-database performance has also been observed.

  12. α-Solanine Modulates the Radiosensitivity of Esophageal Cancer Cells by Inducing MicroRNA 138 Expression

    Directory of Open Access Journals (Sweden)

    Yuanyuan Wang

    2016-08-01

    Full Text Available Background: Esophageal cancer (EC is one of the most common malignant tumors in the world. Due to difficulties with performing the operation, most patients choose to have palliative treatment instead. Radiotherapy is one of the main palliative treatments of EC. However, the clinical efficacy of radiotherapy is not satisfactory α-Solanine is a bioactive component of steroidal glycoalkaloids which has been demonstrated to exhibit anti-metastasis activity in different cancers. In the present study, we determined the effect of α-solanine on the radiosensitivity of EC cells and priliminarily explored the underlying molecular mechanisms. Methods: Cell Counting Kit-8 (CCK-8 assay was conducted to found the cytotoxic effect of α-solanine on EC cells. CCK-8 assay and colony-forming survival assays were performed to explore the effect of α-solanine on cell viability and proliferation of EC cells after irradiation. Immunofluorescence and comet assays were used to detect the effect of α-solanine on DNA repair capacity of EC cells after irradiation. The flow cytometry (FCM and Hoechst/PI staining were conductd to study the effect of α-solanine on apoptosis of EC cells after irradiation. Results: The cytotoxic effect of α-solanine to EC cells was dose-dependent. The results of CCK-8, colony-forming survival assay, immunofluorescence, comet assay, FCM and Hoechst/PI staining showed that α-solanine could enhance the radiosensitivity of EC cells. α-Solanine could downregulate Survivin expression level by upregulating miR-138 expression in EC cells. Upregulation of miR-138 and knock down Survivin both enhanced the radiosensitivity of EC cells. Moreover, Survivin could restore the effect of α-solanine and miR-138 on radiosensitivity of EC cells. Conclusions: α-solanine could enhance the radiosensitivity of esophageal cancer cells by inducing microRNA-138 expression, and probably be an effective radiosensitizer in treating EC.

  13. Human survivin and Trypanosoma cruzi calreticulin act in synergy against a murine melanoma in vivo.

    Directory of Open Access Journals (Sweden)

    Lorena Aguilar-Guzmán

    Full Text Available Immune-based anti-tumor or anti-angiogenic therapies hold considerable promise for the treatment of cancer. The first approach seeks to activate tumor antigen-specific T lymphocytes while, the second, delays tumor growth by interfering with blood supply. Tumor Associated Antigens are often employed to target tumors with therapeutic drugs, but some are also essential for tumor viability. Survivin (Surv is a member of the inhibitor of apoptosis protein family that is considered a Tumor Associated Antigen important for cancer cell viability and proliferation. On the other hand, Trypanosoma cruzi (the agent of Chagas' disease calreticulin (TcCRT displays remarkable anti-angiogenic properties. Because these molecules are associated with different tumor targets, we reasoned that immunization with a Surv-encoding plasmid (pSurv and concomitant TcCRT administration should generate a stronger anti-tumor response than application of either treatment separately. To evaluate this possibility, C57BL/6 mice were immunized with pSurv and challenged with an isogenic melanoma cell line that had been pre-incubated with recombinant TcCRT (rTcCRT. Following tumor cell inoculation, mice were injected with additional doses of rTcCRT. For the combined regimen we observed in mice that: i. Tumor growth was impaired, ii. Humoral anti-rTcCRT immunity was induced and, iii. In vitro rTcCRT bound to melanocytes, thereby promoting the incorporation of human C1q and subsequent macrophage phagocytosis of tumor cells. These observations are interpreted to reflect the consequence of the following sequence of events: rTcCRT anti-angiogenic activity leads to stress in tumor cells. Murine CRT is then translocated to the external membrane where, together with rTcCRT, complement C1 is captured, thus promoting tumor phagocytosis. Presentation of the Tumor Associated Antigen Surv induces the adaptive anti-tumor immunity and, independently, mediates anti-endothelial cell immunity leading

  14. Expression profile of apoptotic and proliferative proteins in hypoxic HUVEC treated with statins.

    Science.gov (United States)

    Li, Xiaochen; Liu, Xiansheng; Xu, Yongjian; He, Yuanzhou; Liu, Jin; Xie, Min

    2015-02-01

    Vascular endothelial hyperproliferation is involved in the pathophysiological process of angiogenesis, which is indispensable for tumor growth and spread in hypoxic adaptation. There is increasing evidence indicating that statins have potential anti-angiogenesis benefits. However, the intracellular signaling mechanism underlying the effect of statins in vascular endothelial cells is undefined. The present study was conducted to investigate the effect of fluvastatin on cell proliferation and apoptosis in normoxic and hypoxic human umbilical vein endothelial cells (HUVEC). Flow cytometric analyses revealed that statins reversed hypoxia-induced cell proliferation by slowing down G1 to S transition and inducing cell apoptosis. To get further insights into the downstream effects of statins, we measured the expression of various apoptosis-associated proteins in hypoxic HUVEC using human apoptosis antibody array. The results suggested that cell apoptosis was accompanied by upregulation of caspase-3, p27, IGFBP-6 and a decrease of bcl-2, survivin levels. Subsequent studies confirmed the results of array and demonstrated that fluvastatin activated mitochondrial apoptosis through enhancing bax/bcl-2 ratio, releasing cytochrome c, in turn activating caspase-9 and caspase-3, and eventually cleaving PARP. Further experiments showed that inhibition of cell proliferation by fluvastatin was associated with elevated IGFBP-6, p27, p53 levels and reduced survivin, cyclin B1, cyclin D1 and VEGF expression. Taken together, fluvastatin suppressed cell proliferation and induced apoptosis of HUVEC in hypoxia via multiple signaling pathways, providing a theoretical basis for statins in the therapy of cancer.

  15. Nitric oxide synthase expression and enzymatic activity in multiple sclerosis

    DEFF Research Database (Denmark)

    Broholm, H; Andersen, B; Wanscher, B

    2004-01-01

    We used post-mortem magnetic resonance imaging (MRI) guidance to obtain paired biopsies from the brains of four patients with clinical definite multiple sclerosis (MS). Samples were analyzed for the immunoreactivity (IR) of the three nitric oxide (NO) synthase isoforms [inducible, neuronal...... and endothelial nitric oxide synthase (NOS)], and enzymatic NO synthase activity. MRI guided biopsies documented more active plaques than macroscopic examination, and histological examination revealed further lesions. Inducible NOS (iNOS) was the dominant IR isoform, while reactive astrocytes were the dominant i......NOS expressing cells in active lesions. NOS IR expressing cells were widely distributed in plaques, in white and gray matter that appeared normal macroscopically, and on MR. Endothelial NOS (eNOS) was highly expressed in intraparenchymal vascular endothelial cells of MS patients. A control group matched for age...

  16. Downregulation of sulfotransferase expression and activity in diseased human livers.

    Science.gov (United States)

    Yalcin, Emine B; More, Vijay; Neira, Karissa L; Lu, Zhenqiang James; Cherrington, Nathan J; Slitt, Angela L; King, Roberta S

    2013-09-01

    Sulfotransferase (SULT) function has been well studied in healthy human subjects by quantifying mRNA and protein expression and determining enzyme activity with probe substrates. However, it is not well known if sulfotransferase activity changes in metabolic and liver disease, such as diabetes, steatosis, or cirrhosis. Sulfotransferases have significant roles in the regulation of hormones and excretion of xenobiotics. In the present study of normal subjects with nonfatty livers and patients with steatosis, diabetic cirrhosis, and alcoholic cirrhosis, we sought to determine SULT1A1, SULT2A1, SULT1E1, and SULT1A3 activity and mRNA and protein expression in human liver tissue. In general, sulfotransferase activity decreased significantly with severity of liver disease from steatosis to cirrhosis. Specifically, SULT1A1 and SULT1A3 activities were lower in disease states relative to nonfatty tissues. Alcoholic cirrhotic tissues further contained lower SULT1A1 and 1A3 activities than those affected by either of the two other disease states. SULT2A1, on the other hand, was only reduced in alcoholic cirrhotic tissues. SULT1E1 was reduced both in diabetic cirrhosis and in alcoholic cirrhosis tissues, relative to nonfatty liver tissues. In conclusion, the reduced levels of sulfotransferase expression and activity in diseased versus nondiseased liver tissue may alter the metabolism and disposition of xenobiotics and affect homeostasis of endobiotic sulfotransferase substrates.

  17. Expression of protooncogenes during lymphocyte activation by growth factors.

    Science.gov (United States)

    Bulanova, E G; Budagyan, V M; Yarilin, A A; Mazurenko, N N

    1997-09-01

    Effects of growth factors of non-immune origin including somatotropin (ST) and platelet-derived growth factor (PDGF) on the expression of the proteins encoded by c-fos, c-myc, c-fun, and c-ets family protooncogenes were studied for the first time. The dynamics of the oncoprotein expression in activated CD(3+)-lymphocytes was investigated by immunoblotting. The accumulation of the Fos and Myc proteins was enhanced in T-lymphocytes treated with ST, PDGF, or phytohemagglutinin; the accumulation was maximum at 30-60 min and decreased in 2 h; the data indicate that the oncoproteins participate in the early lymphocyte activation by various growth factors. The Jun protein appears only in 3 h after the onset of lymphocyte activation; this suggests independent participation of Fos in the early stages of lymphocyte activation prior to the appearance of Jun, preceding the joint action of Fos and Jun within the AP-1 transcription complex. The products of the c-ets family are differentially activated by the studied growth factors. Resting lymphocytes actively accumulate the Ets-1 protein; ST and PDGF activation decreases Ets-1 expression in 2 h. The Ets-2 protein is not detected in resting cells and PDGF-activated lymphocytes, whereas lymphocyte activation by ST is associated with accumulation of Ets-2. The data suggest that the product of the c-ets-1 gene is more important in the regulation of resting cells and the product of the c-ets-2 gene is important during activation of lymphocytes by ST. The results indicate that activation of lymphocytes with growth factors of non-immune origin is mediated by several signal transduction pathways.

  18. Preliminary Study on the Anti-tumor Effect of HLA-A2 .1 Restricted Survivin-△Ex3 -derived CTL Epitope Gene Vaccine%Survivin-△Ex3蛋白HLA-A2.1限制性CTL表位的基因疫苗抗肿瘤效果初探

    Institute of Scientific and Technical Information of China (English)

    陈英利; 葛存兴; 刘思岐; 张婷婷; 隋广宇; 曹明月; 常亚娟; 刘欣宇

    2014-01-01

    To predict and synthesize HLA-A2.1 restricted survivin-△Ex3-derived CTL epitope gene vaccine,and discuss the protective effects of oral attenuated salmonella vaccine with epitope gene on the model mice of transplanted hepatic cellular cancer. Epitope gene sequences with highest scores in prediction were connected to establish eukaryotic expression vector of pVAX1 -STEs-EGFP,which was then transferred into the attenuated salmonella to act as oral vaccine.Experimental animals were divided into the control group (un-vaccinated),oral salmonella group ,oral attenuated salmonella group (transfected by pVAX1 plasmid),oral atten-uated salmonella group (transfected by pVAX1 -Survivin -△3 (T34A)-EGFP plasmid)and oral attenuated salmonella group (transfected by pVAX1 -STEs-EGFP plasmid).The mice were given immunization and then injected liver cancer cells subcutane-ously.The size of the mass at the injection site was measured.In 5 groups of the mice,the average diameters of the tumor were respec-tively 15.11 ±2.43 mm,13.70 ±2.97 mm,13.05 ±1.77 mm,7.46 ±2.61 mm and 9.05 ±2.18 mm;Epitope gene vaccine groups have significant difference when compared with the other groups (P<0.01 ).We concluded that HLA-A2.1 restricted surviving-△Ex3-derived CTL epitope gene vaccine can inhibit the proliferation and migration of mice liver cancer cells after oral administration to mice.%预测并合成survivin-△Ex3 HLA-A2.1限制性CTL表位的基因疫苗,探讨携带表位基因的减毒鼠伤寒沙门氏菌口服疫苗对小鼠移植肝癌模型的保护作用。将表位基因序列连接,构建pVAX1-STEs-EGFP真核表达载体,转化入减毒鼠伤寒沙门氏菌作为口服疫苗。实验动物分为未免疫对照组、口服沙门氏菌组、pVAX1质粒转染的减毒鼠伤寒沙门氏菌组;口服pVAX1-Survivin-△3(T34A)! EGFP质粒转染的的减毒鼠伤寒沙门氏菌组和口服pVAX1-STEs ! EGFP质粒转染的的减毒鼠伤寒沙门

  19. Lotus hairy roots expressing inducible arginine decarboxylase activity.

    Science.gov (United States)

    Chiesa, María A; Ruiz, Oscar A; Sánchez, Diego H

    2004-05-01

    Biotechnological uses of plant cell-tissue culture usually rely on constitutive transgene expression. However, such expression of transgenes may not always be desirable. In those cases, the use of an inducible promoter could be an alternative approach. To test this hypothesis, we developed two binary vectors harboring a stress-inducible promoter from Arabidopsis thaliana, driving the beta-glucuronidase reporter gene and the oat arginine decarboxylase. Transgenic hairy roots of Lotus corniculatus were obtained with osmotic- and cold-inducible beta-glucuronidase and arginine decarboxylase activities. The increase in the activity of the latter was accompanied by a significant rise in total free polyamines level. Through an organogenesis process, we obtained L. corniculatus transgenic plants avoiding deleterious phenotypes frequently associated with the constitutive over-expression of arginine decarboxylation and putrescine accumulation.

  20. Activation of peroxisome proliferator-activated receptor α in human endothelial cells increases plasminogen activator inhibitor type-1 expression

    Institute of Scientific and Technical Information of China (English)

    叶平; 胡晓晖; 刘永学; 赵亚力

    2003-01-01

    Objective To investigate the effect of peroxisome proliferator-activated receptors (PPARs) activators on plasminogen activator inhibitor 1 (PAI-1) expression in human umbilical vein endothelial cells and elucidate a possible mechanism.Methods Human umbilical vein endothelial cells (HUVECs) were obtained from normal fetus, and cultured conventionally. Then the HUVEC were exposed to fatty acids and prostaglandin J2 in varying concentrations with fresh media. RT-PCR and ELISA were used to determine the expression of PPAR and PAI-1 in HUVECs. Transient co-transfection of PAI-1 promoter and PPARα gene or PPARγ gene to ECV304 was performed.Results PPARα, PPARδ and PPARγ mRNA in HUVECs were detected by RT-PCR. Treatment of HUVECs with PPARα and PPARγ activators-linolenic acid, linoleic acid, oleic acid and prostaglandin J2, but not with stearic acid could augment PAI-I mRNA expression and protein secretion in a concentration-dependent manner. Proportional induction of PAI-1 promoter activity was observed through increasing amounts of PPARα DNA in HUVECs through a transient gene transfection assay, although the mRNA expression of the 3 subtypes of PPAR with their activators were not changed compared with controls.Conclusions HUVECs express PPARs. PPARs activators may increase PAI-1 expression in endothelial cells (EC). Although PPARs expression was not enhanced after being stimulated by their activators in EC, the functionally active PPARα is probably involved in regulating PAI-1 expression in EC.

  1. Pre-Vaccination Frequencies of Th17 Cells Correlate with Vaccine-Induced T-Cell Responses to Survivin-Derived Peptide Epitopes

    DEFF Research Database (Denmark)

    Køllgaard, Tania; Ugurel-Becker, Selma; Idorn, Manja

    2015-01-01

    Various subsets of immune regulatory cells are suggested to influence the outcome of therapeutic antigen-specific anti-tumor vaccinations. We performed an exploratory analysis of a possible correlation of pre-vaccination Th17 cells, MDSCs, and Tregs with both vaccination-induced T-cell responses...... an altered activity of immune regulatory cells. Moreover, the frequencies of Th17 cells (p=0.03) and Tregs (p=0.02) were elevated as compared to healthy donors. IL-17-secreting CD4+ T cells displayed an impact on the immunological and clinical effects of vaccination: Patients characterized by high...... frequencies of Th17 cells at pre-vaccination were more likely to develop survivin-specific T-cell reactivity post-vaccination (p=0.03). Furthermore, the frequency of Th17 (p=0.09) and Th17/IFNγ+ (p=0.19) cells associated with patient survival after vaccination. In summary, our explorative, hypothesis...

  2. Prefrontal parvalbumin interneurons shape neuronal activity to drive fear expression.

    Science.gov (United States)

    Courtin, Julien; Chaudun, Fabrice; Rozeske, Robert R; Karalis, Nikolaos; Gonzalez-Campo, Cecilia; Wurtz, Hélène; Abdi, Azzedine; Baufreton, Jerome; Bienvenu, Thomas C M; Herry, Cyril

    2014-01-02

    Synchronization of spiking activity in neuronal networks is a fundamental process that enables the precise transmission of information to drive behavioural responses. In cortical areas, synchronization of principal-neuron spiking activity is an effective mechanism for information coding that is regulated by GABA (γ-aminobutyric acid)-ergic interneurons through the generation of neuronal oscillations. Although neuronal synchrony has been demonstrated to be crucial for sensory, motor and cognitive processing, it has not been investigated at the level of defined circuits involved in the control of emotional behaviour. Converging evidence indicates that fear behaviour is regulated by the dorsomedial prefrontal cortex (dmPFC). This control over fear behaviour relies on the activation of specific prefrontal projections to the basolateral complex of the amygdala (BLA), a structure that encodes associative fear memories. However, it remains to be established how the precise temporal control of fear behaviour is achieved at the level of prefrontal circuits. Here we use single-unit recordings and optogenetic manipulations in behaving mice to show that fear expression is causally related to the phasic inhibition of prefrontal parvalbumin interneurons (PVINs). Inhibition of PVIN activity disinhibits prefrontal projection neurons and synchronizes their firing by resetting local theta oscillations, leading to fear expression. Our results identify two complementary neuronal mechanisms mediated by PVINs that precisely coordinate and enhance the neuronal activity of prefrontal projection neurons to drive fear expression.

  3. [Comparison of expression and antibacterial activities of recombinant porcine lactoferrin expressed in four Lactobacillus species].

    Science.gov (United States)

    Yu, Hui; Jiang, Yanping; Cui, Wen; Wu, Xiao; He, Jia; Qiao, Xinyuan; Li, Yijing; Tang, Lijie

    2014-09-01

    The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.

  4. Diverse intracellular pathogens activate type III interferon expression from peroxisomes.

    Science.gov (United States)

    Odendall, Charlotte; Dixit, Evelyn; Stavru, Fabrizia; Bierne, Helene; Franz, Kate M; Durbin, Ann Fiegen; Boulant, Steeve; Gehrke, Lee; Cossart, Pascale; Kagan, Jonathan C

    2014-08-01

    Type I interferon responses are considered the primary means by which viral infections are controlled in mammals. Despite this view, several pathogens activate antiviral responses in the absence of type I interferons. The mechanisms controlling type I interferon-independent responses are undefined. We found that RIG-I like receptors (RLRs) induce type III interferon expression in a variety of human cell types, and identified factors that differentially regulate expression of type I and type III interferons. We identified peroxisomes as a primary site of initiation of type III interferon expression, and revealed that the process of intestinal epithelial cell differentiation upregulates peroxisome biogenesis and promotes robust type III interferon responses in human cells. These findings highlight the importance of different intracellular organelles in specific innate immune responses.

  5. Clinical study on diagnostic utility of Survivin-molecular beacons in bladder cancer%Survivin分子信标诊断膀胱肿瘤的临床应用研究

    Institute of Scientific and Technical Information of China (English)

    王新阳; 贺大林; 杨小杰; 赵军

    2013-01-01

    Objective:To develop a sensitive method for detection of bladder cancer cells the cast-off cells of bladder cancer patients,we examine the feasibility by using molecular beacon (MB) probes specific for a tumor specific Survivin mRNA.Methods:MB analyzed the survivn mRNA in bladder cancer cell 5637,J82 and identified by Western blot.Urine cytology,MB test,and Western blot were carried to test,cancer tissue of 35 bladder cancer patients and 35 health adults.Results:Survivin MB could detect the expression of Survivin gene and generated fluorescent signals in the cancer cells,and not detect in normal prostate fibroblast cells.MB detected cancerous cells in 80% of confirmed bladder cancer patients (28/35).The specificity was 77.1% (27/35).Survivin protein was detected by western blot in 71.4% (25/35) of these patients.The two methods had high consistency.The sensitivity and specificity of urine cytology was 28.6% (10/35),100% (35/35).Conclusion:Survivin MB is specific and sensitive molecular probe for detecting bladder cancer cells and urine cast-off cells of bladder cancer patients.It has great potential for the development of a clinical diagnostic procedure for early detection of bladder cancer and follow-up after operation.%目的:探讨分子信标检测尿脱落细胞Survivn mRNA的可行性,寻找一种能够早期诊断膀胱肿瘤的方法.方法:分子信标检测膀胱肿瘤5637、J82细胞Survivin mRNA的表达,并通过Western bolt方法验证,并对35例膀胱移行细胞癌患者和35名正常健康成人行分子信标检测尿脱落细胞,Western bolt检测组织中的Survivin含量,同时行尿脱落细胞学检查.结果:Survivin分子信标检测肿瘤细胞内的Survivin表达且具有高特异性.以随机100个细胞中60个以上的细胞为阳性做为阳性标准,确定MB-cy3的阳性率为80% (28/35),特异性为77.1%(27/35);Western bolt检测的阳性率为71.4%(25/35).两种实验方法对细胞和蛋白质中Survivin的检

  6. Guiding individualized therapy in advanced non-small cell lung cancer by detection of Survivin protein%检测Survivin蛋白指导NSCLC的个体化治疗

    Institute of Scientific and Technical Information of China (English)

    朱建荣; 郁震; 万小蔹; 崔小川

    2015-01-01

    目的 探讨根据肿瘤组织中Survivin蛋白的表达情况指导非小细胞肺癌(NSCLC)进行个体化治疗的作用和意义.方法 117例确诊为NSCLC的患者按2∶1比例随机分2组.个体化治疗组(n=78)根据组织标本中Survivin蛋白的表达情况,选择个体化方案化疗,Survivin蛋白阳性患者采用非铂化疗方案,Survivin蛋白阴性患者采用含铂类药物化疗方案;标准治疗组(n=39)选择含铂类药物的一线标准方案化疗.比较2个治疗组的化疗效果,以Kaplan-Meier法分析2个治疗组患者生存期的差异.结果 NSCLC患者中Survivin蛋白表达的阳性率为51.3%;个体化治疗组和标准治疗组的化疗有效率分别为55.1%和33.3%(x2=4.949,P=0.026),两组的平均生存期分别为13.7个月和10.8个月,个体化治疗组的疗效明显优于标准治疗组(P =0.009).结论 检测Survivin蛋白以指导NSCLC患者进行个体化治疗可以提高化疗效果,并延长生存时间.%Objective To investigate the value of detecting Survivin protein in biopsy specimens of advanced non-small cell lung cancer (NSCLC) patients for individualized therapy.Methods 117 pathologically proven NSCLC patients were enrolled and randomly assigned in 2 ∶ 1 ratio to either the individualized treatment group or the standard treatment group.In individualized treatment group,platinum-based chemotherapy was given to Survivin protein negative patients,chemotherapy without platinum was given to Survivin protein positive patients after Survivin assessment.The standard treatment group received first-line platinum-based chemotherapy regimen.Differences in treatment effect between the groups were statistically analyzed.Survival differences were analyzed by Kaplan-Meier survival curves.Results The expression of Survivin protein was 51.3% among 78 cases.There was statistical significance in outcome between the individualized treatment group and the standard treatment group (response rate:55.1% vs 33.3

  7. Expression of Telomerase Activity in Gastric Cancer Cells

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objective To study the relationship between telomerase activity and biological behavior in human gastric cells and appraise the clinical significance of detecting telomerase activity. Methods The telomerase activity in 47 gastric cancer tissue samples,their matched nomal tissues,7 gastric ulcer and 2 gastric cancer cell lines was detected using a PCR-based non-radioisotopic telomeric repeat amplification protocol(TRAP) assay. Results None of the 47 samples from normal gastric tissues expressed telomerase activity.The 41 of 47 cases of gastric cancer presented telomerase activity with an 87.2% positive rate (P<0.001). 2/2 gastric cancer cell lines and 0/7 gastric ulcer line were also positive for telmerase activity.The activity of telomerase was associated with the pathological differentiation of gastric cancer. Conclusion Telomerase activity may be related to the biological behavior of gastric cancer and can help in assessing the malignant poten-tial of gastric cancer.Telomerase activity will be a good diagnostic marker for the detection of gastric cancer.

  8. Pre-Vaccination Frequencies of Th17 Cells Correlate with Vaccine-Induced T-Cell Responses to Survivin-Derived Peptide Epitopes.

    Science.gov (United States)

    Køllgaard, Tania; Ugurel-Becker, Selma; Idorn, Manja; Andersen, Mads Hald; Becker, Jürgen C; Straten, Per Thor

    2015-01-01

    Various subsets of immune regulatory cells are suggested to influence the outcome of therapeutic antigen-specific anti-tumor vaccinations. We performed an exploratory analysis of a possible correlation of pre-vaccination Th17 cells, MDSCs, and Tregs with both vaccination-induced T-cell responses as well as clinical outcome in metastatic melanoma patients vaccinated with survivin-derived peptides. Notably, we observed dysfunctional Th1 and cytotoxic T cells, i.e. down-regulation of the CD3ζchain (p=0.001) and an impaired IFNγ-production (p=0.001) in patients compared to healthy donors, suggesting an altered activity of immune regulatory cells. Moreover, the frequencies of Th17 cells (p=0.03) and Tregs (p=0.02) were elevated as compared to healthy donors. IL-17-secreting CD4+ T cells displayed an impact on the immunological and clinical effects of vaccination: Patients characterized by high frequencies of Th17 cells at pre-vaccination were more likely to develop survivin-specific T-cell reactivity post-vaccination (p=0.03). Furthermore, the frequency of Th17 (p=0.09) and Th17/IFNγ+ (p=0.19) cells associated with patient survival after vaccination. In summary, our explorative, hypothesis-generating study demonstrated that immune regulatory cells, in particular Th17 cells, play a relevant role for generation of the vaccine-induced anti-tumor immunity in cancer patients, hence warranting further investigation to test for validity as predictive biomarkers.

  9. Adaptation of muscle gene expression to changes in contractile activity

    Science.gov (United States)

    Booth, F. W.; Babij, P.; Thomason, D. B.; Wong, T. S.; Morrison, P. R.

    1987-01-01

    A review of the existing literature regarding the effects of different types of physical activities on the gene expression of adult skeletal muscles leads us to conclude that each type of exercise training program has, as a result, a different phenotype, which means that there are multiple mechanisms, each producing a unique phenotype. A portion of the facts which support this position is presented and interpreted here. [Abstract translated from the original French by NASA].

  10. TNF gene expression in macrophage activation and endotoxin tolerance

    OpenAIRE

    Chow, Nancy Ann-Marie

    2013-01-01

    TNF is an inflammatory cytokine that plays a critical role in the acute phase response to infection, and its dysregulation has been implicated in the pathology of several inflammatory and autoimmune disorders. TNF gene expression is regulated in a cell type- and inducer-specific manner that involves chromatin alterations at both the TNF promoter and distal DNase I hypersensitive (DH) sites within the TNF/LT locus. While the mechanisms underlying TNF gene activation in monocytes/macrophages an...

  11. Inhibition of STAT3 Expression and Signaling in Resveratrol-Differentiated Medulloblastoma Cells

    Directory of Open Access Journals (Sweden)

    Li-Jun Yu

    2008-07-01

    Full Text Available In this study, the potential influence of resveratrol (3,5,4′-trihydroxy-trans-stilbene in signal transducer and activator of transcription 3 (STAT3 signaling of medulloblastoma cells was evaluated by checking the status of STAT3 signaling and its downstream gene expression in two medulloblastoma cell lines (UW228-2 and UW228-3 with and without resveratrol treatment. The results revealed that resveratrol induced neuronal differentiation of medulloblastoma cells. Signal transducer and activator of transcription 3 expression and phosphorylation were detected in normally cultured UW228-2 and UW228-3 cells that were apparently attenuated after resveratrol treatment. The expression of STAT3 downstream genes, survivin, cyclin D1, Cox-2, and c-Myc, was suppressed but Bcl-2 was enhanced by resveratrol. Meanwhile, the production and secretion of leukemia inhibitory factor, a STAT3 activator, became active in resveratrol-treated cells. To further ascertain the significance of STAT3 signaling for medulloblastoma cells, AG490, a selective inhibitor of STAT3 phosphorylation, was used to treat UW228-3 cells. Phosphorylation of STAT3 was inhibited by AG490 accompanied with growth suppression, differentiation-like changes, and down-regulation of survivin, cyclin D1, Cox-2, and c-Myc. Our data thus suggest the importance of STAT3 signaling in maintenance and survival of medulloblastoma cells. This signaling may be the major target of resveratrol. Enhanced leukemia inhibitory factor and Bcl-2 expressions in resveratrol-treated cells might reflect a compensatory response to the loss of STAT3 function.

  12. Expression and activity of a novel cathelicidin from domestic cats.

    Directory of Open Access Journals (Sweden)

    Brian C Leonard

    Full Text Available Cathelicidins are small cationic antimicrobial peptides found in many species including primates, mammals, marsupials, birds and even more primitive vertebrates, such as the hagfish. Some animals encode multiple cathelicidins in their genome, whereas others have only one. This report identifies and characterizes feline cathelicidin (feCath as the sole cathelicidin in domestic cats (Felis catus. Expression of feCath is predominantly found in the bone marrow, with lower levels of expression in the gastrointestinal tract and skin. By immunocytochemistry, feCath localizes to the cytoplasm of neutrophils in feline peripheral blood. Structurally, the mature feCath sequence is most similar to a subgroup of cathelicidins that form linear α-helices. feCath possesses antimicrobial activity against E. coli D31, Salmonella enterica serovar Typhimurium (IR715, Listeria monocytogenes and Staphylococcus pseudintermedius (clinical isolate similar to that of the human ortholog, LL-37. In contrast, feCath lacks the DNA binding activity seen with LL-37. Given its similarity in sequence, structure, tissue expression, and antimicrobial activity, the cathelicidin encoded by cats, feCath, belongs to the subgroup of linear cathelicidins found not only in humans, but also non-human primates, dogs, mice, and rats.

  13. Expression of biologically active murine interleukin-18 in Lactococcus lactis.

    Science.gov (United States)

    Feizollahzadeh, Sadegh; Khanahmad, Hossein; Rahimmanesh, Ilnaz; Ganjalikhani-Hakemi, Mazdak; Andalib, Alireza; Sanei, Mohammad Hossein; Rezaei, Abbas

    2016-11-01

    The food-grade bacterium Lactococcus lactis is increasingly used for heterologous protein expression in therapeutic and industrial applications. The ability of L. lactis to secrete biologically active cytokines may be used for the generation of therapeutic cytokines. Interleukin (IL)-18 enhances the immune response, especially on mucosal surfaces, emphasizing its therapeutic potential. However, it is produced as an inactive precursor and has to be enzymatically cleaved for maturation. We genetically manipulated L. lactis to secrete murine IL-18. The mature murine IL-18 gene was inserted downstream of a nisin promoter in pNZ8149 plasmid and the construct was used to transform L. lactis NZ3900. The transformants were selected on Elliker agar and confirmed by restriction enzyme digestion and sequencing. The expression and secretion of IL-18 protein was verified by SDS-PAGE, western blotting and ELISA. The biological activity of recombinant IL-18 was determined by its ability to induce interferon (IFN)-γ production in L. lactis co-cultured with murine splenic T cells. The amounts of IL-18 in bacterial lysates and supernatants were 3-4 μg mL(-1) and 0.6-0.7 ng mL(-1), respectively. The successfully generated L. lactis strain that expressed biologically active murine IL-18 can be used to evaluate the possible therapeutic effects of IL-18 on mucosal surfaces.

  14. Regulation of gene expression in ovarian cancer cells by luteinizing hormone receptor expression and activation

    Directory of Open Access Journals (Sweden)

    Dam Phuongan

    2011-06-01

    Full Text Available Abstract Background Since a substantial percentage of ovarian cancers express gonadotropin receptors and are responsive to the relatively high concentrations of pituitary gonadotropins during the postmenopausal years, it has been suggested that receptor activation may contribute to the etiology and/or progression of the neoplasm. The goal of the present study was to develop a cell model to determine the impact of luteinizing hormone (LH receptor (LHR expression and LH-mediated LHR activation on gene expression and thus obtain insights into the mechanism of gonadotropin action on ovarian surface epithelial (OSE carcinoma cells. Methods The human ovarian cancer cell line, SKOV-3, was stably transfected to express functional LHR and incubated with LH for various periods of time (0-20 hours. Transcriptomic profiling was performed on these cells to identify LHR expression/activation-dependent changes in gene expression levels and pathways by microarray and qRT-PCR analyses. Results Through comparative analysis on the LHR-transfected SKOV-3 cells exposed to LH, we observed the differential expression of 1,783 genes in response to LH treatment, among which five significant families were enriched, including those of growth factors, translation regulators, transporters, G-protein coupled receptors, and ligand-dependent nuclear receptors. The most highly induced early and intermediate responses were found to occupy a network impacting transcriptional regulation, cell growth, apoptosis, and multiple signaling transductions, giving indications of LH-induced apoptosis and cell growth inhibition through the significant changes in, for example, tumor necrosis factor, Jun and many others, supportive of the observed cell growth reduction in in vitro assays. However, other observations, e.g. the substantial up-regulation of the genes encoding the endothelin-1 subtype A receptor, stromal cell-derived factor 1, and insulin-like growth factor II, all of which are

  15. 生存蛋白与口腔疾病间的关系%Relationship between survivin and oral diseases

    Institute of Scientific and Technical Information of China (English)

    李倩(综述); 潘亚萍(审校)

    2013-01-01

    Survivin,?a?member?of?the?inhibitor?of?apoptosis?protein?family,?functions?as?a?regulator?of?mitosis?and?cell-death?decisions.?Survivin?affects?the?initiation?and?progression?of?diseases?by?interfering?with?cell?proliferation?and?apoptosis.?In?recent?years,?the?relationship?between?survivin?and?oral?diseases?has?attracted?considerable?research?attention.?This?article?focuses?on?the?biological?functions?of?survivin?as?well?as?relevant?research?progress?in?oral?cancer,?periodontal?diseases,?and?salivary?gland?development.%生存蛋白(survivin)具有调控细胞有丝分裂和抑制程序性细胞死亡的双重功能,可干扰细胞的增殖与程序性死亡,进而影响疾病的发生发展进程。近年来,生存蛋白与口腔疾病间的关系备受瞩目,本文就生存蛋白及其与口腔肿瘤、牙周病和唾液腺发育间的关系等研究进展作一综述。

  16. Pharmacological and Genetic Modulation of REV-ERB Activity and Expression Affects Orexigenic Gene Expression.

    Directory of Open Access Journals (Sweden)

    Ariadna Amador

    Full Text Available The nuclear receptors REV-ERBα and REV-ERBβ are transcription factors that play pivotal roles in the regulation of the circadian rhythm and various metabolic processes. The circadian rhythm is an endogenous mechanism, which generates entrainable biological changes that follow a 24-hour period. It regulates a number of physiological processes, including sleep/wakeful cycles and feeding behaviors. We recently demonstrated that REV-ERB-specific small molecules affect sleep and anxiety. The orexinergic system also plays a significant role in mammalian physiology and behavior, including the regulation of sleep and food intake. Importantly, orexin genes are expressed in a circadian manner. Given these overlaps in function and circadian expression, we wanted to determine whether the REV-ERBs might regulate orexin. We found that acute in vivo modulation of REV-ERB activity, with the REV-ERB-specific synthetic ligand SR9009, affects the circadian expression of orexinergic genes in mice. Long term dosing with SR9009 also suppresses orexinergic gene expression in mice. Finally, REV-ERBβ-deficient mice present with increased orexinergic transcripts. These data suggest that the REV-ERBs may be involved in the repression of orexinergic gene expression.

  17. Signaling lymphocyte activating molecule (SLAM) expression in subacute sclerosing panencephalitis.

    Science.gov (United States)

    Piskin, A Kevser; Akpinar, Pinar; Muftuoglu, Sevda; Anlar, Banu

    2007-08-01

    Signaling lymphocyte activating molecule (SLAM) is a receptor for measles virus which also has immunomodulatory activity. We analyzed SLAM expression in mononuclear cells (MNC) of patients with SSPE (n=7) and control subjects (n=7) from the same population. Native 10% PAGE analysis in cell and brain tissue extracts followed by Western blotting using monoclonal anti-human SLAM showed four types of bands. Differences in the type and amount of SLAM expression were observed between SSPE and control cases. Lymphocytes of SSPE patients showed two types of SLAM bands in comparison to only one in control lymphocytes. Stimulation of cells with lipopolysaccharide (80 u/ml) and concanavalin A (1 microg/ml) in vitro led to the appearance of a second isoform in both groups. Brain homogenates of SSPE patients (n=2) displayed all four types of SLAM isoforms at significantly higher levels than those of control brains (n=2). Our results show native PAGE enables the detection of all SLAM isotypes. The expression of SLAM is increased in lymphocytes, monocytes, and brain tissues of SSPE patients.

  18. PTEN基因对慢性粒细胞白血病Survivin、Xiap、Smac调控的研究%Regulation of wild type PTEN gene on Survivin, Xiap and Smac in chronic leukemia cells

    Institute of Scientific and Technical Information of China (English)

    成志勇; 万建设; 王亚丽; 梁丽青; 梁文同; 穆敬; 芦希; 潘崚

    2011-01-01

    Objective To explore the effects of tumor-suppressing gene wild type PTEN on the cell proliferation,apoptosis and the possible regulations of apoptosis-related molecules Survivin,Xiap and Smac gene in human chronic myeloid leukemia(CML)and cell line K562 cells.Methods(1)The recombinant adenovirus containing green fluorescent protein(GFP)and PTEN(Ad-PTEN-GFP)or empty vector(AdGFP)was transfected into K562 cells.The growth of K562 cells was observed by MTT assay while cell cycle and apoptotic rate were assessed by flow cytometry(FCM).PTEN,Survivin,Xiap and Smac mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR(FQ-PCR)while PTEN protein levels analyzed by Western blot.(2)The expression levels of PTEN,Survivin,Xiap and Smac mRNA were detected in 10 chronic myelogenous leukemia(CML)patients in chronic phase(CML-CP),10 CML patients in blast crises(CML-BC)and 10 normal control marrow mononuclear cells(MMNC).Results The growth of K562 cells was suppressed markedly.And the maximal growth inhibition rate was 38.6% after the tranfection of PTEN.Survivin,Xiap,Smac mRNA expression levels were down-regulated by around 6.14,7.44 and 2.95 folds respectively(0.0700 ±0.0059,0.0089 ±0.0006,0.0600 ±0.0039 vs 0.4370 ± 0.0790,0.0661 ± 0.0072,0.1580 ± 0.0078 vs 0.4530 ± 0.0810,0.0700 ± 0.0079.0.1770 ±0.0085,all P < 0.01).The mRNA expression level of PTEN in CML-BC patients was lower than that in CML-CP patients and normal control.But Survivin,Xiap,Smae mRNA expression levels were higher in CML-BC patients than those in CML-CP and normal control.Conclusion The over-expression of PTEN gene may inhibit the proliferation of K562 cells and promote cell apoptosis via the regulation of Survivin,Xiap and Smac genes.%目的 探讨肿瘤抑制基因PTEN对人慢性粒细胞白血病(CML)中生存素(Survivin)、X连锁凋亡抑制蛋白(Xiap)、线粒体促凋亡蛋白(Smac)调控的作用.方法(1)将携带有野生型PTEN和绿

  19. Expression, Purification and Anti-tumor Activity of Curcin

    Institute of Scientific and Technical Information of China (English)

    Meng-Jun LUO; Xin-Yu YANG; Wei-Xin LIU; Ying XU; Ping HUANG; Fang YAN; Fang CHEN

    2006-01-01

    Curcin, purified from the seeds of Jatropha curcas, can be used as a cell-killing agent. Understanding the anti-tumor activity of the recombinant protein of curcin is important for its application in clinical medicine.The segment encoding the mature protein of curcin was inserted into Escherichia coli strain M15, and the recombinant strain was induced to express by isopropyl-β-D-thiogalactopyranoside at concentration of 0.5 mM. The recombinant protein was expressed in the form of inclusion bodies and purified by Ni-NTA affinity chromatography. The target protein was incubated with the tumor cells at different concentrations for different times and the results demonstrated that the target protein could inhibit the growth of tumor cells (NCL-H446, SGC-7901 and S180) at 5 μg/ml.

  20. Behavioral meaningful opioidergic stimulation activates kappa receptor gene expression

    Energy Technology Data Exchange (ETDEWEB)

    Teodorov, E. [Centro de Matemática, Computação e Cognição, Universidade Federal do ABC, São Paulo, SP (Brazil); Ferrari, M.F.R. [Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, São Paulo, SP (Brazil); Fior-Chadi, D.R. [Departamento de Fisiologia, Instituto de Biociências, Universidade de São Paulo, São Paulo, SP (Brazil); Camarini, R. [Departamento de Farmacologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP (Brazil); Felício, L.F. [Departamento de Patologia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP (Brazil)

    2012-06-01

    The periaqueductal gray (PAG) has been reported to be a location for opioid regulation of pain and a potential site for behavioral selection in females. Opioid-mediated behavioral and physiological responses differ according to the activity of opioid receptor subtypes. The present study investigated the effects of the peripheral injection of the kappa-opioid receptor agonist U69593 into the dorsal subcutaneous region of animals on maternal behavior and on Oprk1 gene activity in the PAG of female rats. Female Wistar rats weighing 200-250 g at the beginning of the study were randomly divided into 2 groups for maternal behavior and gene expression experiments. On day 5, pups were removed at 7:00 am and placed in another home cage that was distant from their mother. Thirty minutes after removing the pups, the dams were treated with U69593 (0.15 mg/kg, sc) or 0.9% saline (up to 1 mL/kg) and after 30 min were evaluated in the maternal behavior test. Latencies in seconds for pup retrieval, grouping, crouching, and full maternal behavior were scored. The results showed that U69593 administration inhibited maternal behavior (P < 0.05) because a lower percentage of U69593 group dams showed retrieval of first pup, retrieving all pups, grouping, crouching and displaying full maternal behavior compared to the saline group. Opioid gene expression was evaluated using real-time reverse-transcription polymerase chain reaction (RT-PCR). A single injection of U69593 increased Oprk1 PAG expression in both virgin (P < 0.05) and lactating female rats (P < 0.01), with no significant effect on Oprm1 or Oprd1 gene activity. Thus, the expression of kappa-opioid receptors in the PAG may be modulated by single opioid receptor stimulation and behavioral meaningful opioidergic transmission in the adult female might occur simultaneously to specific changes in gene expression of kappa-opioid receptor subtype. This is yet another alert for the complex role of the opioid system in female

  1. Behavioral meaningful opioidergic stimulation activates kappa receptor gene expression

    Directory of Open Access Journals (Sweden)

    E. Teodorov

    2012-10-01

    Full Text Available The periaqueductal gray (PAG has been reported to be a location for opioid regulation of pain and a potential site for behavioral selection in females. Opioid-mediated behavioral and physiological responses differ according to the activity of opioid receptor subtypes. The present study investigated the effects of the peripheral injection of the kappa-opioid receptor agonist U69593 into the dorsal subcutaneous region of animals on maternal behavior and on Oprk1 gene activity in the PAG of female rats. Female Wistar rats weighing 200-250 g at the beginning of the study were randomly divided into 2 groups for maternal behavior and gene expression experiments. On day 5, pups were removed at 7:00 am and placed in another home cage that was distant from their mother. Thirty minutes after removing the pups, the dams were treated with U69593 (0.15 mg/kg, sc or 0.9% saline (up to 1 mL/kg and after 30 min were evaluated in the maternal behavior test. Latencies in seconds for pup retrieval, grouping, crouching, and full maternal behavior were scored. The results showed that U69593 administration inhibited maternal behavior (P < 0.05 because a lower percentage of kappa group dams showed retrieval of first pup, retrieving all pups, grouping, crouching and displaying full maternal behavior compared to the saline group. Opioid gene expression was evaluated using real-time reverse-transcription polymerase chain reaction (RT-PCR. A single injection of U69593 increased Oprk1 PAG expression in both virgin (P < 0.05 and lactating female rats (P < 0.01, with no significant effect on Oprm1 or Oprd1 gene activity. Thus, the expression of kappa-opioid receptors in the PAG may be modulated by single opioid receptor stimulation and behavioral meaningful opioidergic transmission in the adult female might occur simultaneously to specific changes in gene expression of kappa-opioid receptor subtype. This is yet another alert for the complex role of the opioid system in

  2. The Value of Combined Use of Survivin mRNA and Matrix Metalloproteinase 2 and 9 for Bladder Cancer Detection in Voided Urine

    Directory of Open Access Journals (Sweden)

    Sanaa Eissa

    2013-01-01

    Full Text Available Objective: In a trial to improve the diagnostic efficacy of conventional urine cytology we determine survivin RNA and matrix metalloproteinase 2 and 9 in urine of bladder cancer cases.

  3. 口腔黏膜下纤维化癌变过程中G2、M期细胞周期蛋白与存活素磷酸化的研究%Molecules of G2/M phase and the phosphorylation of survivin in the carcinogenesis of oral submucosal fibrosis

    Institute of Scientific and Technical Information of China (English)

    周晌辉; 李力力; 翦新春; 王颖; 陈新群; 高兴

    2008-01-01

    目的 探讨细胞周期G2、M期重要分子细胞周期蛋白B1(Cyclin B1)、细胞分裂周期蛋白2(cell division cycle 2,p34cdc2)和存活素磷酸化在口腔黏膜下纤维化(oral submucosa fibrosis,OSF)癌变过程中的作用与意义.方法 应用Western blotting检测10例正常口腔黏膜上皮组织、40例OSF上皮组织及42例OSF癌变组织中Cyclin B1、p34cdc2和存活素磷酸化的表达情况,免疫共沉淀实验分析p34cdc2激酶与存活素的相关性.结果 Cyclin B1、p34cdc2、磷酸化p34cdc(p-p34cdc2)和存活素磷酸化在OSF组织中表达显著高于正常口腔黏膜(P<0.05);在OSF癌变组织中的表达显著高于OSF组织(P<0.05);但在OSF早、中、晚期3组间差异无统计学意义(P>0.05).免疫共沉淀实验证实了p34cdc与存活素的结合.结论 细胞周期G2、M期重要分子Cyclin B1、p34cdc2及存活素磷酸化在OSF细胞分裂增殖过程中起促进作用,对OSF癌变的早期诊断和治疗具有重要的临床意义.%Objective To determine the expression of Cyclin B1,p34cdc2 and the phosphorylation of survivin(p-survivin)in oral squamous cell carcinoma and oral submucosa fibrosis(OSF),and to discuss their possible role in carcinogenesis of OSF.Methods The expression of Cyclin B1,p34cdc2 and p-survivin were analyzed by Western blotting assay in 10 cases of normal oral mucosa epithelium,40 cases of OSF epithelium and 42 cases of oral squamous cell carcinoma(OSCC)originated from OSF,respectively.Immunoprecipitation was used to confirrn the relationship between the p34cdc2 and survivin.Results The expression of Cyclin B1,p34cdc2,p-p34cdc2 and p-survivin in OSF group were significantly higher than those in normal group(P<0.05).The expression of these molecules showed significant different(P<0.05)between the OSF and OSCC originated from OSF,but there was no significant difference among the early stage,the moderately advanced stage and the advanced stage of OSF.Immunoprecipitation assay

  4. Brain Activity while Reading Sentences with Kanji Characters Expressing Emotions

    Science.gov (United States)

    Yuasa, Masahide; Saito, Keiichi; Mukawa, Naoki

    In this paper, we describe the brain activity associated with kanji characters expressing emotion, which are places at the end of a sentence. Japanese people use a special kanji character in brackets at the end of sentences in text messages such as those sent through e-mail and messenger tools. Such kanji characters plays a role to expresses the sender's emotion (such as fun, laughter, sadness, tears), like emoticons. It is a very simple and effective way to convey the senders' emotions and his/her thoughts to the receiver. In this research, we investigate the effects of emotional kanji characters by using an fMRI study. The experimental results show that both the right and left inferior frontal gyrus, which have been implicated on verbal and nonverbal information, were activated. We found that we detect a sentence with an emotional kanji character as the verbal and nonverval information, and a sentence with emotional kanji characters enrich communication between the sender and the reciever.

  5. Activated Expression of WRKY57 Confers Drought Tolerance in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Yanjuan Jiang; Gang Liang; Diqiu Yu

    2012-01-01

    Drought is one of the most serious environmental factors that limit the productivity of agricultural crops worldwide.However,the mechanism underlying drought tolerance in plants is unclear.WRKY transcription factors are known to function in adaptation to abiotic stresses.By screening a pool of WRKY-associated T-DNA insertion mutants,we isolated a gain-of-function mutant,acquired drought tolerance (adt),showing improved drought tolerance.Under drought stress conditions,adt accumulated higher levels of ABA than wild-type plants.Stomatal aperture analysis indicated that adt was more sensitive to ABA than wild-type plants.Molecular genetic analysis revealed that a T-DNA insertion in adt led to activated expression of a WRKY gene that encodes the WRKR57 protein.Constitutive expression of WRKY57 also conferred similar drought tolerance.Consistently with the high ABA content and enhanced drought tolerance,three stress-responsive genes (RD29A,NCED3,and ABA3) were up-regulated in adt.ChIP assays demonstrated that WRKY57 can directly bind the W-box of RD29A and NCED3 promoter sequences.In addition,during ABA treatment,seed germination and early seedling growth of adt were inhibited,whereas,under high osmotic conditions,adt showed a higher seed germination frequency.In summary,our results suggested that the activated expression of WRKY57 improved drought tolerance of Arabidopsis by elevation of ABA levels.Establishment of the functions of WRKY57 will enable improvement of plant drought tolerance through gene manipulation approaches.

  6. Sulindac metabolites inhibit epidermal growth factor receptor activation and expression

    Directory of Open Access Journals (Sweden)

    Pangburn Heather A

    2005-09-01

    Full Text Available Abstract Background Regular use of nonsteroidal anti-inflammatory drugs (NSAIDs is associated with a decreased mortality from colorectal cancer (CRC. NSAIDs induce apoptotic cell death in colon cancer cells in vitro and inhibit growth of neoplastic colonic mucosa in vivo however, the biochemical mechanisms required for these growth inhibitory effects are not well defined. We previously reported that metabolites of the NSAID sulindac downregulate extracellular-signal regulated kinase 1/2 (ERK1/2 signaling and that this effect is both necessary and sufficient for the apoptotic effects of these drugs. The goal of this project was to specifically test the hypothesis that sulindac metabolites block activation and/or expression of the epidermal growth factor (EGF receptor (EGFR. Methods HT29 human colon cancer cells were treated with EGF, alone, or in the presence of sulindac sulfide or sulindac sulfone. Cells lysates were assayed by immunoblotting for phosphorylated EGFR (pEGFR, pY1068, total EGFR, phosphorylated ERK1/2 (pERK1/2, total ERK1/2, activated caspase-3, and α-tubulin. Results EGF treatment rapidly induced phosphorylation of both EGFR and ERK1/2 in HT29 colon cancer cells. Pretreatment with sulindac metabolites for 24 h blocked EGF-induced phosphorylation of both EGFR and ERK1/2 and decreased total EGFR protein expression. Under basal conditions, downregulation of pEGFR and total EGFR was detected as early as 12 h following sulindac sulfide treatment and persisted through at least 48 h. Sulindac sulfone induced downregulation of pEGFR and total EGFR was detected as early as 1 h and 24 h, respectively, following drug treatment, and persisted through at least 72 h. EGFR downregulation by sulindac metabolites was observed in three different CRC cell lines, occurred prior to the observed downregulation of pERK1/2 and induction of apoptosis by these drugs, and was not dependent of caspase activation. Conclusion These results suggest that

  7. Evaluation of the Antioxidant Activity and Antiproliferative Effect of the Jaboticaba (Myrciaria cauliflora Seed Extracts in Oral Carcinoma Cells

    Directory of Open Access Journals (Sweden)

    Wen-Hung Wang

    2014-01-01

    Full Text Available It is becoming increasingly evident that certain phytochemicals possess cancer chemopreventive properties. In this study, the antiproliferative activity of extracts from different parts of the jaboticaba (Myrciaria cauliflora plant was evaluated for its effect on human oral carcinoma cell lines. The cytotoxicities of various plant extract concentrations were examined and the 50% maximal inhibitory concentration (IC50 was determined. Water extracts of jaboticaba seeds showed concentration-dependent antiproliferative effects. Annexin V/propidium iodide positivity with active caspase-3 induction indicated that the treated cells underwent apoptosis. Several important regulatory proteins (Bcl-2, Bcl-xL, Bid, and survivin involved in apoptosis were also evaluated. The antioxidant activity of jaboticaba was investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS assays, and the drug concentration eliciting 50% maximum stimulation (SC50 was determined. The present findings suggest that water extracts of jaboticaba seeds exhibit an antiproliferative effect against oral cancer cells by inducing apoptosis through downregulating survivin expression and thereby activating caspase-mediated Bid cleavage.

  8. Efficient expression and purification of biologically active human cystatin proteins.

    Science.gov (United States)

    Chauhan, Sakshi; Tomar, Raghuvir S

    2016-02-01

    Cystatins are reversible cysteine protease inhibitor proteins. They are known to play important roles in controlling cathepsins, neurodegenerative disease, and in immune system regulation. Production of recombinant cystatin proteins is important for biochemical and function characterization. In this study, we cloned and expressed human stefin A, stefin B and cystatin C in Escherichia coli. Human stefin A, stefin B and cystatin C were purified from soluble fraction. For cystatin C, we used various chaperone plasmids to make cystatin C soluble, as it is reported to localize in inclusion bodies. Trigger factor, GroES-GroEL, DnaK-DnaJ-GrpE chaperones lead to the presence of cystatin C in the soluble fraction. Immobilized metal affinity chromatography, glutathione sepharose and anion exchange chromatography techniques were employed for efficient purification of these proteins. Their biological activities were tested by inhibition assays against cathepsin L and H3 protease.

  9. Synergistic Inhibitory Effect of survivin siRNA in Combination with 5-Fu on Inhibiting Proliferation of MCF-7 Cells%靶向survivin siRNA与5-Fu协同抑制MCF-7细胞增殖

    Institute of Scientific and Technical Information of China (English)

    薛兴欢; 张淑群; 姜建涛; 王西京; 薛锋杰; 刘晓旭

    2008-01-01

    目的 研究以survivin为靶标的小干扰RNA(siRNA)与化疗药5-Fu联合应用抑制MCF-7细胞增殖的作用.方法 以脂质体为载体,将survivin siRNA转染至MCF-7细胞中,用四氮唑盐(MTT)法染色并计算siRNA联用5-Fu对MCF-7细胞的抑制率,用SAS统计软件及金正均Q值法进行统计分析.结果 单用5-Fu,TC50为4.42μg/ml;加入5 nmol/L siRNA后,IC50降为1.18μg/ml;siRNA与5-Fu联用的抑制作用较单用5-Fu强(F=26.74,P<0.01);Q值分析表明survivin siRNA与中低浓度的5-Fu联用,有较好的协同作用(Q≥1.15).结论 survivin siRNA与5-Fu联用,可显著增强对MCF-7细胞增殖的抑制,提高肿瘤细胞对化疗药物的敏感性.

  10. Noscapine Induced Apoptosis via Downregulation of Survivin in Human Neuroblastoma Cells Having Wild Type or Null p53

    OpenAIRE

    Shiwang Li; Jing He; Shuai Li; Guoqing Cao; Shaotao Tang; Qiangsong Tong; Joshi, Harish C.

    2012-01-01

    Neuroblastoma is the most common extracranial solid tumor of childhood. It accounts for 15% of pediatric cancer deaths. Chemotherapy is the mainstay of treatment in children with advanced neuroblastoma. Noscapine, a nontoxic natural compound, can trigger apoptosis in many cancer types. We now show that p53 is dispensable for Noscapine-induced cell death in neuroblastoma cell lines, proapoptotic response to this promising chemopreventive agent is mediated by suppression of survivin protein exp...

  11. Exspression of STAT3, VEGF and survivin in gastric carcinoma%信号传导与转录活化因子3、血管内皮生长因子和Survivin在胃癌组织中的表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    刘翔宇; 邓靖宇; 张霖; 梁寒

    2010-01-01

    Objective To evaluate the expression of STAT3, VEGF and Survivin in human gastric carcinoma and its clinicopathological significance. Methods The expression of STAT3, VEGF, survivin was determined by immunohistochemical staining of specimens from 53 cases undergoing radical gastrectomy and 53 cases of normal gastric mucous membranae. We evaluated the relationship between expression of these proteins and various clinicopothological factors. Results The expression rate of STAT3, VEGF and survivin in 53 gastric carcinoma tissues was 58%, 62% and 74%, respectively, which was significantly higher than those in the normal group(P <0. 01). STAT3 expression correlated with VEGF(r =0. 608 ,P <0. 01) ,survivin(r = 0. 451, P = 0. 001). Positive STAT3, VEGF staining was significantly associated with tumor size, Lauren's classification,lymph node metastasis and clinical staging(P < 0. 05). Survivin staining was significantly associated with Lauren's classification, lymph node metastasis and clinical staging(P <0. 05). Multivariate analysis revealed STAT3 expression and lymph node metastasis were independently prognostic factors of poor survival. Conclusion VEGF, survivin possibly regulated by STAT3 leads to tumor angiogenesis and anti-apoptosis. The expression of STAT3 is an independent prognostic factors in gastric carcinoma.%目的 探讨信号传导与转录活化因子3(STAT3)、血管内皮生长因子(VEGF)和Survivin蛋白在胃癌组织中的表达情况以及三者与胃癌临床病理特征的关系.方法 用免疫组织化学方法检测53例行胃癌根治切除的胃癌组织和53例正常胃组织中STAT3、VEGF和Survivin蛋白表达情况,并且分析3种蛋白表达与胃癌生物学行为的关系.结果 在本组53例胃癌标本中,STAT3、VEGF和Survivin蛋白的阳性表达率分别为58%、62%、74%,明显高于正常胃组织(P<0.01).胃癌组织中STAT3与VEGF、Survivin蛋白的阳性表达呈正相关(r=0.608、0.451,P<0.01),STAT3、VEGF的表达与

  12. Piezometric biosensors for anti-apoptotic protein survivin based on buried positive-potential barrier and immobilized monoclonal antibodies.

    Science.gov (United States)

    Stobiecka, Magdalena; Chalupa, Agata; Dworakowska, Beata

    2016-10-15

    The anti-apoptotic protein survivin (Sur) plays an important role in the regulation of cell division and inducing the chemotherapeutic drug resistance. The Sur protein and its mRNA have recently been studied as cancer biomarkers and potential targets for cancer therapy. In this work, we have focused on the design of immunosensors for the detection of Sur based on buried positive-potential barrier layer structure and anti-survivin antibody. The modification of solid AuQC piezoelectrodes was monitored by recording the resonance frequency shift and electrochemical measurements during each step of the sensor preparation. Our results indicate that the immunosensor with covalently bound monoclonal anti-survivin antibody can detect Sur with the limit of detection, LOD=1.7nM (S/N=3σ). The immunosensor applicability for the analysis of real samples was assessed by testing samples of cell lysate solutions obtained from human astrocytoma (glioblastoma) U-87MG cell line, with the experiments performed using the standard addition method. The good linearity of the calibration curves for PBS and lysate solutions at low Sur concentrations confirm the high specificity of the proposed biosensor and good discrimination against nonspecific interactions with lysate components. The calculations indicate that there is still room to increase the Sur capture capacity for Sur while miniaturizing the sensor. The important advantage of the sensor is that it can be reused by a simple regeneration procedure.

  13. Xenoestrogenic gene expression: structural features of active polycyclic aromatic hydrocarbons.

    Science.gov (United States)

    Schultz, T Wayne; Sinks, Glendon D

    2002-04-01

    Estrogenicity was assessed using the Saccharomyces cerevisiae-based Lac-Z reporter assay and was reported as the logarithm of the inverse of the 50% molar beta-galactosidase activity (log[EC50(-1)]). In an effort to quantify the relationship between molecular structure of polycyclic aromatic hydrocarbons (PAHs) and estrogenic gene expression, a series of PAHs were evaluated. With noted exceptions, the results of these studies indicate that the initial two-dimensional structural warning for estrogenicity, the superpositioning of a hydroxylated aromatic system on the phenolic A-ring of 17-beta-estradiol, can be extended to the PAHs. This two-dimensional-alignment criterion correctly identified estrogenicity of 22 of the 29 PAHs evaluated. Moreover, the estrogenic potency of these compounds was directly related to the size of the hydrophobic backbone. The seven compounds classified incorrectly by this structural feature were either dihydroxylated naphthalenes or aromatic nitrogen-heterocyclic compounds; all such compounds were false positives. Results with dihydroxylated naphthalenes reveal derivatives that were nonestrogenic when superimposed on the phenolic A-ring of 17-beta-estradiol had the second hydroxyl group in the position of the C-ring or were catechol-like in structure. Structural alerts for nitrogen-heterocyclic compounds must take into account the position of the hydroxyl group and the in-ring nitrogen atom; compounds with the hydroxyl group and nitrogen atom involved with the same ring were observed to be nonactive.

  14. Opinions expressed by Italian National Advisory Toxicological Committee on some active ingredients of pesticides

    Energy Technology Data Exchange (ETDEWEB)

    Camoni, I. [Ist. Superiore di Sanita`, Rome (Italy). Lab. di Tossicologia Applicata

    1996-03-01

    The opinions expressed by the Italian National Advisory Toxicological Committee (CCTN) on some active ingredients of pesticides are presented. Carcinogenic and mutagenic effects of these substances have been examined and, on this basis, an evaluation and relative classification were expressed.

  15. Clinical Development of Gamitrinib, a Novel Mitochondrial-Targeted Small Molecule Hsp90 Inhibitor

    Science.gov (United States)

    2015-09-01

    trophoblast, an actively invasive tissue at the interface between fetal and maternal circulation [30], whereas expression of this molecule in the adult...chondrial survivin (3). Furthermore, reconstitution of survivin-depleted PC3 cells with adenovirus ( pAd ) encoding mitochondrial-targeted survivin (3... pAd - mt-SVV) stimulated O2 consumption (Fig. 2G). In contrast, PC3 cells transfected with nontargeting siRNA and reconstituted with pAd -mt-survivin

  16. Arabidopsis TTG2 regulates TRY expression through enhancement of activator complex-triggered activation.

    Science.gov (United States)

    Pesch, Martina; Dartan, Burcu; Birkenbihl, Rainer; Somssich, Imre E; Hülskamp, Martin

    2014-10-01

    Trichome patterning in Arabidopsis thaliana is regulated by a regulatory feedback loop of the trichome promoting factors TRANSPARENT TESTA GLABRA1 (TTG1), GLABRA3 (GL3)/ENHANCER OF GL3 (EGL3), and GL1 and a group of homologous R3MYB proteins that act as their inhibitors. Together, they regulate the temporal and spatial expression of GL2 and TTG2, which are considered to control trichome cell differentiation. In this work, we show that TTG2 is a specific activator of TRY (but not CPC or GL2). The WRKY protein TTG2 binds to W-boxes in a minimal promoter fragment of TRY, and these W-boxes are essential for rescue of the try mutant phenotype. We further show that TTG2 alone is not able to activate TRY expression, but rather drastically enhances the activation by TTG1 and GL3. As TTG2 physically interacts with TTG1 and because TTG2 can associate with GL3 through its interaction with TTG1, we propose that TTG2 enhances the activity of TTG1 and GL3 by forming a protein complex.

  17. Expression of IAP family proteins and its clinical importance in breast cancer patients.

    Science.gov (United States)

    Pluta, P; Jeziorski, A; Cebula-Obrzut, A Pluta B; Wierzbowska, A; Piekarski, J; Smolewski, P

    2015-01-01

    Inhibitor of apoptosis (IAP) family proteins is involved in mechanisms of resistance to apoptosis in various cancer cells. The aim of this study was to assess the expression of selected IAP proteins such as XIAP, cIAP-1, cIAP-2 and survivin in breast cancer patients and evaluates their relationship with the prognostic and predictive factors and their impact to overall survival (OS) and progression free survival (PFS). The study was conducted with the use of tissue samples prospectively collected from 92 previously untreated female breast cancer patients. The control encompassed 10 fibroadenoma patients. The expression of XIAP, cIAP-1, cIAP-2 and survivin was assessed using flow multicolor cytometry. XIAP expression was present in 99 % of the breast cancer patients (91/92) with the median expression 13.65% (range 1-66.8%). Expression of XIAP in breast cancer was significantly higher compared to the control group (p=0.006). Median expression of cIAP-1, cIAP-2 and survivin in the study group was 25.95% (range 0.8-83.7%), 16.7% (range 1-53.2%) and 4.6% (range 0-43%) respectively. In the rank Spearman test, strong correlations (pproteins and survival. However, low expression of XIAP in breast cancer showed trend to longer PFS (p=0.08). XIAP, cIAP-1 cIAP-2 and survivin participate in antiapoptotic mechanisms in breast cancer and XIAP and survivin seem to have the most significant prognostic importance. Further studies are needed to establish more complete prognostic and predictive values of IAP family proteins in breast cancer patients.

  18. Structural Basis for Recognition of H3T3ph and Smac/DIABLO N-terminal Peptides by Human Survivin

    Energy Technology Data Exchange (ETDEWEB)

    Du, Jiamu; Kelly, Alexander E.; Funabiki, Hironori; Patel, Dinshaw J. (MSKCC); (Rockefeller)

    2012-03-02

    Survivin is an inhibitor of apoptosis family protein implicated in apoptosis and mitosis. In apoptosis, it has been shown to recognize the Smac/DIABLO protein. It is also a component of the chromosomal passenger complex, a key player during mitosis. Recently, Survivin was identified in vitro and in vivo as the direct binding partner for phosphorylated Thr3 on histone H3 (H3T3ph). We have undertaken structural and binding studies to investigate the molecular basis underlying recognition of H3T3ph and Smac/DIABLO N-terminal peptides by Survivin. Our crystallographic studies establish recognition of N-terminal Ala in both complexes and identify intermolecular hydrogen-bonding interactions in the Survivin phosphate-binding pocket that contribute to H3T3ph mark recognition. In addition, our calorimetric data establish that Survivin binds tighter to the H3T3ph-containing peptide relative to the N-terminal Smac/DIABLO peptide, and this preference can be reversed through structure-guided mutations that increase the hydrophobicity of the phosphate-binding pocket.

  19. Anger expression and natural killer cell activity in family caregivers participating in a physical activity trial.

    Science.gov (United States)

    Wilcox, S; King, A C; Vitaliano, P P; Brassington, G S

    2000-07-01

    Associations between psychological functioning and natural killer cell activity (NKA) were examined in 23 older (62.2 ± 7.5 years) family caregivers randomized to a moderate intensity four-month exercise program or to a wait-list control condition. At baseline, although NKA was related to anger-control (r = -.42; trend p caregiver burden. After controlling for baseline NKA, changes in anger-control explained 14 percent of the variance in NKA four months later. Decreases in anger-control predicted increases in NKA. Group assignment (exercise vs control) was unrelated to changes in NKA over the four-month period; however, the study was not powered to detect this effect. These results are consistent with reported relationships of anger expression with other physiological measures, and extend the importance of anger expression to immune functioning in older family caregivers.

  20. α-Tomatine-mediated anti-cancer activity in vitro and in vivo through cell cycle- and caspase-independent pathways.

    Directory of Open Access Journals (Sweden)

    Min-Wu Chao

    Full Text Available α-Tomatine, a tomato glycoalkaloid, has been reported to possess antibiotic properties against human pathogens. However, the mechanism of its action against leukemia remains unclear. In this study, the therapeutic potential of α-tomatine against leukemic cells was evaluated in vitro and in vivo. Cell viability experiments showed that α-tomatine had significant cytotoxic effects on the human leukemia cancer cell lines HL60 and K562, and the cells were found to be in the Annexin V-positive/propidium iodide-negative phase of cell death. In addition, α-tomatine induced both HL60 and K562 cell apoptosis in a cell cycle- and caspase-independent manner. α-Tomatine exposure led to a loss of the mitochrondrial membrane potential, and this finding was consistent with that observed on activation of the Bak and Mcl-1 short form (Mcl-1s proteins. Exposure to α-tomatine also triggered the release of the apoptosis-inducing factor (AIF from the mitochondria into the nucleus and down-regulated survivin expression. Furthermore, α-tomatine significantly inhibited HL60 xenograft tumor growth without causing loss of body weight in severe combined immunodeficiency (SCID mice. Immunohistochemical test showed that the reduced tumor growth in the α-tomatine-treated mice was a result of increased apoptosis, which was associated with increased translocation of AIF in the nucleus and decreased survivin expression ex vivo. These results suggest that α-tomatine may be a candidate for leukemia treatment.

  1. Transgenic mice expressing constitutive active MAPKAPK5 display gender-dependent differences in exploration and activity

    Directory of Open Access Journals (Sweden)

    Moens Ugo

    2007-11-01

    Full Text Available Abstract Background The mitogen-activated protein kinases, MAPKs for short, constitute cascades of signalling pathways involved in the regulation of several cellular processes that include cell proliferation, differentiation and motility. They also intervene in neurological processes like fear conditioning and memory. Since little remains known about the MAPK-Activated Protein Kinase, MAPKAPK5, we constructed the first MAPKAPK knockin mouse model, using a constitutive active variant of MAPKAPK5 and analyzed the resulting mice for changes in anxiety-related behaviour. Methods We performed primary SHIRPA observations during background breeding into the C57BL/6 background and assessed the behaviour of the background-bred animals on the elevated plus maze and in the light-dark test. Our results were analyzed using Chi-square tests and homo- and heteroscedatic T-tests. Results Female transgenic mice displayed increased amounts of head dips and open arm time on the maze, compared to littermate controls. In addition, they also explored further into the open arm on the elevated plus maze and were less active in the closed arm compared to littermate controls. Male transgenic mice displayed no differences in anxiety, but their locomotor activity increased compared to non-transgenic littermates. Conclusion Our results revealed anxiety-related traits and locomotor differences between transgenic mice expressing constitutive active MAPKAPK5 and control littermates.

  2. Building gene expression signatures indicative of transcription factor activation to predict AOP modulation

    Science.gov (United States)

    Building gene expression signatures indicative of transcription factor activation to predict AOP modulation Adverse outcome pathways (AOPs) are a framework for predicting quantitative relationships between molecular initiatin...

  3. Detecting microRNA activity from gene expression data

    LENUS (Irish Health Repository)

    Madden, Stephen F

    2010-05-18

    Abstract Background MicroRNAs (miRNAs) are non-coding RNAs that regulate gene expression by binding to the messenger RNA (mRNA) of protein coding genes. They control gene expression by either inhibiting translation or inducing mRNA degradation. A number of computational techniques have been developed to identify the targets of miRNAs. In this study we used predicted miRNA-gene interactions to analyse mRNA gene expression microarray data to predict miRNAs associated with particular diseases or conditions. Results Here we combine correspondence analysis, between group analysis and co-inertia analysis (CIA) to determine which miRNAs are associated with differences in gene expression levels in microarray data sets. Using a database of miRNA target predictions from TargetScan, TargetScanS, PicTar4way PicTar5way, and miRanda and combining these data with gene expression levels from sets of microarrays, this method produces a ranked list of miRNAs associated with a specified split in samples. We applied this to three different microarray datasets, a papillary thyroid carcinoma dataset, an in-house dataset of lipopolysaccharide treated mouse macrophages, and a multi-tissue dataset. In each case we were able to identified miRNAs of biological importance. Conclusions We describe a technique to integrate gene expression data and miRNA target predictions from multiple sources.

  4. Detecting microRNA activity from gene expression data.

    LENUS (Irish Health Repository)

    Madden, Stephen F

    2010-01-01

    BACKGROUND: MicroRNAs (miRNAs) are non-coding RNAs that regulate gene expression by binding to the messenger RNA (mRNA) of protein coding genes. They control gene expression by either inhibiting translation or inducing mRNA degradation. A number of computational techniques have been developed to identify the targets of miRNAs. In this study we used predicted miRNA-gene interactions to analyse mRNA gene expression microarray data to predict miRNAs associated with particular diseases or conditions. RESULTS: Here we combine correspondence analysis, between group analysis and co-inertia analysis (CIA) to determine which miRNAs are associated with differences in gene expression levels in microarray data sets. Using a database of miRNA target predictions from TargetScan, TargetScanS, PicTar4way PicTar5way, and miRanda and combining these data with gene expression levels from sets of microarrays, this method produces a ranked list of miRNAs associated with a specified split in samples. We applied this to three different microarray datasets, a papillary thyroid carcinoma dataset, an in-house dataset of lipopolysaccharide treated mouse macrophages, and a multi-tissue dataset. In each case we were able to identified miRNAs of biological importance. CONCLUSIONS: We describe a technique to integrate gene expression data and miRNA target predictions from multiple sources.

  5. Apoptosis of HL-60 human leukemia cells induced by Asiatic acid through modulation of B-cell lymphoma 2 family proteins and the mitogen-activated protein kinase signaling pathway.

    Science.gov (United States)

    Wu, Qiuling; Lv, Tingting; Chen, Yan; Wen, Lu; Zhang, Junli; Jiang, Xudong; Liu, Fang

    2015-07-01

    The toxicities of conventional chemotherapeutic agents to normal cells restrict their dosage and clinical efficacy in acute leukemia; therefore, it is important to develop novel chemotherapeutics, including natural products, which selectively target cancer-specific pathways. The present study aimed to explore the effect of the chemopreventive agent asiatic acid (AA) on the proliferation and apoptotic rate of the leukemia cell line HL-60 and investigated the mechanisms underlying its anti-tumor activity. The effect of AA on the proliferation of HL-60 cells was evaluated using the MTT assay. Annexin V-fluorescein isothiocyanate/propidium iodide double staining followed by flow cytometric analysis as well as Hoechst 33258 staining were used to analyze the apoptotic rate of the cells. Furthermore, changes of survivin, B-cell lymphoma 2 (Bcl-2), myeloid cell leukemia 1 (Mcl-1), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 expressions were detected by western blot analysis. AA blocked the growth of HL-60 cells in a dose- and time-dependent manner. The IC50-value of AA on HL-60 cells was 46.67 ± 5.08 µmol/l for 24 h. AA induced apoptosis in a dose-dependent manner, which was inhibited in the presence of Z-DEVD-FMK, a specific inhibitor of caspase. The anti-apoptotic proteins Bcl-2, Mcl-1 and survivin were downregulated by AA in a dose-dependent manner. Concurrently, AA inhibited ERK and p38 phosphorylation in a dose-dependent manner, while JNK phosphorylation was not affected. In conclusion, the present study indicated that the p38 and ERK pathways, as well as modulation of Bcl-2 family and survivin proteins were key regulators of apoptosis induced in HL-60 cells in response to AA.

  6. Immune Killing Activity of Lymphocytes on Hela Cells Expressing Interleukin-12 In Vitro

    Institute of Scientific and Technical Information of China (English)

    Huiyan WANG; Suhua CHEN

    2008-01-01

    The killing effects of lymphocytes on Hela cells expressing intedeukin-12 (IL-12) in vitro were explored. By using gene transfection technique, full length IL-12 gene was transfected into Hela cells. The expression of IL-12 in Hela cells was detected quantitatively by ELISA; Changes in killing effects of lymphocytes on Hela cells expressing IL-12 were observed by MTT. It was found that Hela cells could express IL-12 between 24h and 72h after transfection. Killing activity of lymphocytes on Hela cells expressing IL-12 was significantly enhanced. It was concluded by cell transfection technique, Hela cells could express IL-12 and were more easily killed by lymphocytes.

  7. Curcumin inhibits srebp-2 expression in activated hepatic stellate cells in vitro by reducing the activity of specificity protein-1.

    Science.gov (United States)

    Kang, Qiaohua; Chen, Anping

    2009-12-01

    Elevated levels of cholesterol/low-density lipoprotein (LDL) are a risk factor for the development of nonalcoholic steatohepatitis and its associated hepatic fibrosis. However, underlying mechanisms remain elusive. We previously reported that curcumin induced gene expression of peroxisome proliferator-activated receptor (PPAR)-gamma and stimulated its activity, leading to the inhibition of the activation of hepatic stellate cells (HSCs), the major effector cells during hepatic fibrogenesis. We recently showed that curcumin suppressed gene expression of LDL receptor in activated HSCs in vitro by repressing gene expression of the transcription factor sterol regulatory element binding protein-2 (SREBP-2), leading to the reduction in the level of intracellular cholesterol in HSCs and to the attenuation of the stimulatory effects of LDL on HSCs activation. The current study aimed at exploring molecular mechanisms by which curcumin inhibits srebp-2 expression in HSCs. Promoter deletion assays, mutagenesis assays, and EMSAs localize a specificity protein-1 (SP-1) binding GC-box in the srebp-2 promoter, which is responsible for enhancing the promoter activity and responding to curcumin in HSCs. Curcumin suppresses gene expression of SP-1 and reduces its trans-activation activity, which are mediated by the activation of PPARgamma. The inhibitory effect of curcumin on SP-1 binding to the GC-box is confirmed by chromatin immuno-precipitation. In summary, our results demonstrate that curcumin inhibits srebp-2 expression in cultured HSCs by activating PPARgamma and reducing the SP-1 activity, leading to the repression of ldlr expression. These results provide novel insights into molecular mechanisms by which curcumin inhibits LDL-induced HSC activation.

  8. Peroxisome proliferator-activated receptor-g is essential in the pathogenesis of gastric carcinoma

    Institute of Scientific and Technical Information of China (English)

    Xiu-Mei Ma; Hong Yu; Na Huai

    2009-01-01

    AIM: To investigate whether peroxisome proliferatoractivated receptor γ(PPAR-γ) is expressed in human gastric carcinoma and whether PPAR-γ is a potential target for gastric carcinoma therapy. METHODS: PPAR-γ protein in gastric carcinoma was examined by immunohistochemistry. In the gastric carcinoma cell line MGC803, PPAR-γ, survivin, Skp2 and p27 protein and mRNA were examined by Western blotting and real-time reverse transcription-polymerase chain reaction, respectively; proliferation was examined by MTT; apoptosis was examined by chromatin staining with Hoechst 33342 and fluorescence activated cell sorting (FACS). and cell cycle was examined by FACS; the knockdown of PPAR-g was done by RNA interference.RESULTS: A high level of expression of PPAR-γ was observed in human gastric carcinoma and in a human gastric carcinoma cell line MGC803. The PPAR-γ agonist 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) inhibited growth, and induced apoptosis and G1/G0 cell cycle arrest in MGC803 cells in a concentration-dependent and time-dependent manner. The effect of 15d-PGJ2 on MGC803 cells was not reversed by the selective and irreversible antagonist GW9662 for PPAR-γ. Furthermore,survivin and Skp2 expression were decreased, whereas p27 expression was enhanced following 15d-PGJ2 treatment in a dose-dependent manner in MGC803 cells.Interestingly, we also found that small interfering RNA for PPAR-γ inhibited growth and induced apoptosis in MGC803 cells. The inhibition of PPAR-γ function may be a potentially important and novel modality for treatment and prevention of gastric carcinoma.CONCLUSION: A PPAR-γ agonist inhibited growth of human gastric carcinoma MGC803 cells by inducing apoptosis and G1/G0 cell cycle arrest with the involvement of survivin, Skp2 and p27 and not via PPAR-γ.

  9. Hepatic cytochrome P450 activity, abundance, and expression throughout human development

    Energy Technology Data Exchange (ETDEWEB)

    Sadler, Natalie C.; Nandhikonda, Premchendar; Webb-Robertson, Bobbie-Jo M.; Ansong, Charles; Anderson, Lindsey N.; Smith, Jordan N.; Corley, Richard A.; Wright, Aaron T.

    2016-07-01

    Cytochrome P450s are Phase I metabolic enzymes that play critical roles in the biotransformation of endogenous compounds and xenobiotics. The expression and activity of P450 enzymes can vary considerably throughout human development, especially when comparing fetal development to neonates, children, and adults. In an effort to develop a more comprehensive understanding of the ontogeny of P450 expression and activity we employed a multi-omic characterization of P450 transcript expression, protein abundance, and functional activity. To quantify the functional activity of individual P450s we employ activity-based protein profiling, which uses modified mechanism-based inhibitors of P450s as chemical probes, in tandem with proteomic analyses to quantify activity. Our results reveal life-stage-dependent variability in P450 expression, abundance, and activity throughout human development and frequent discordant relationships between expression and activity. The results were used to distribute P450s into three general classes based upon developmental stage of expression and activity. We have significantly expanded the knowledge of P450 ontogeny, particularly at the level of individual P450 activity. We anticipate that our ontogeny results will be useful for enabling predictive therapeutic dosing, and for avoiding potentially adverse and harmful reactions during maturation from both therapeutic drugs and environmental xenobiotics.

  10. Knockdown of STAT3 expression by RNAi induces apoptosis in astrocytoma cells

    Directory of Open Access Journals (Sweden)

    Kruger Mathew M

    2003-09-01

    Full Text Available Abstract Background Astrocytomas are the most common type of primary central nervous system tumors. They are frequently associated with genetic mutations that deregulate cell cycle and render these tumors resistant to apoptosis. STAT3, signal transducer and activator of transcription 3, participates in several human cancers by inducing cell proliferation and inhibiting apoptosis and is frequently activated in astrocytomas. Methods RNA interference was used to knockdown STAT3 expression in human astrocytes and astrocytoma cell lines. The effect of STAT3 knockdown on apoptosis, cell proliferation, and gene expression was then assessed by standard methods. Results We have found that STAT3 is constitutively activated in several human astrocytoma cell lines. Knockdown of STAT3 expression by siRNA induces morphologic and biochemical changes consistent with apoptosis in several astrocytoma cell lines, but not in primary human astrocytes. Moreover, STAT3 is required for the expression of the antiapoptotic genes survivin and Bcl-xL in the A172 glioblastoma cell line. Conclusion These results show that STAT3 is required for the survival of some astrocytomas. These studies suggest STAT3 siRNA could be a useful therapeutic agent for the treatment of astrocytomas.

  11. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard

    2009-01-01

    We show that inhibition of HDAC activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC inhibitor-mediated Hsp70 surface expression was confined to the apoptotic Annexin V...... activity selectively induces surface expression of Hsp70 on hematopoietic cancer cells and that this may increase immunorecognition of these cells.......-positive cells and blocked by inhibition of apoptosis. Other chemotherapeutic inducers of apoptosis such as etoposide and camptothecin also led to a robust induction of Hsp70 surface expression. Hsp70 expression was, however, not caused by induction of apoptosis per se, as activated CD4 T cells remained Hsp70...

  12. Lack of correlation between telomere length and telomerase activity and expression in leukemic cells.

    Science.gov (United States)

    Januszkiewicz, Danuta; Wysoki, Jacek; Lewandowski, Krzysztof; Pernak, Monika; Nowicka, Karina; Rembowska, Jolanta; Nowak, Jerzy

    2003-12-01

    The expression of three components of telomerase complex (hTR, hTERT, TP1) along with telomerase activity and telomere length in leukemic cells was investigated. Cells were isolated from peripheral blood and/or bone marrow of children with acute lymphoblastic (ALL) and non-lymphoblastic (ANLL) leukemia. Expression of three components of telomerase as well as telomerase activity was found in all leukemic cells. Chemiluminescent detection of terminal restriction fragments (TRF) from DNA isolated from ALL cells showed variable patterns expressing considerable heterogeneity of telomere length. The ALL cells appeared to have both long and short telomere lengths, in contrast to normal peripheral lymphocytes, which produced limited pattern of TRF. The ANLL cells produced predominantly short telomere pattern despite high telomerase activity and expression. It can be concluded that high telomerase activity and expression in leukemic cells is not always correlated with long telomeres (TRF pattern).

  13. Novel polyacrylate-based cationic nanoparticles for survivin siRNA delivery combined with mitoxantrone for treatment of breast cancer.

    Science.gov (United States)

    Arami, Sanam; Mahdavi, Majid; Rashidi, Mohammad Reza; Fathi, Marziyeh; Hejazi, Mohammad-Saeid; Samadi, Nasser

    2016-11-01

    As a gene delivery method in breast cancer therapy, knocking down the undesired genes in the cancerous cells would be promising. Inhibitors of Apoptosis Protein (IAP) family genes are some of the genes whose responsibility is inhibition of apoptosis in cells. Silencing these genes seems to be helpful directing the tumor cells to death. siRNA sequence designed against survivin anti-apoptotic gene can play this role if carried to the cytoplasm. Here we prepared a positive charged biocompatible nano-sized particle made up of a Fe3O4 core covered respectively by polyacrylate (PA) and polyethyleneimine (PEI) layer, which could successfully deliver the siRNA into the MCF-7 cells. The particle structure was checked and having less than 50 nm diameter in size, positive charge and, safety towards MCF-7 cells besides being able to form nanoplexes with the siRNA strand helps it entering into the biologic assays part. The siRNA delivery evaluated via flowcytometry. Apoptosis induction was determined by DAPI staining. The efficiency of survivin gene knockdown was evaluated in mRNA and protein levels using Real time PCR and western blotting methods. Overall, the Fe3O4-PA-PEI nanoparticles can deliver siRNA effectively into the cytoplasm of the MCF-7 breast cancer cells and induce apoptosis.

  14. Variable epitope library carrying heavily mutated survivin-derived CTL epitope variants as a new class of efficient vaccine immunogen tested in a mouse model of breast cancer.

    Science.gov (United States)

    NoeDominguez-Romero, Allan; Zamora-Alvarado, Rubén; Servín-Blanco, Rodolfo; Pérez-Hernández, Erendira G; Castrillon-Rivera, Laura E; Munguia, Maria Elena; Acero, Gonzalo; Govezensky, Tzipe; Gevorkian, Goar; Manoutcharian, Karen

    2014-01-01

    The antigenic variability of tumor cells leading to dynamic changes in cancer epitope landscape along with escape from immune surveillance by down-regulating tumor antigen expression/presentation and immune tolerance are major obstacles for the design of effective vaccines. We have developed a novel concept for immunogen construction based on introduction of massive mutations within the epitopes targeting antigenically variable pathogens and diseases. Previously, we showed that these immunogens carrying large combinatorial libraries of mutated epitope variants, termed as variable epitope libraries (VELs), induce potent, broad and long lasting CD8+IFN-γ+ T-cell response as well as HIV-neutralizing antibodies. In this proof-of-concept study, we tested immunogenic properties and anti-tumor effects of the VELs bearing survivin-derived CTL epitope (GWEPDDNPI) variants in an aggressive metastatic mouse 4T1 breast tumor model. The constructed VELs had complexities of 10,500 and 8,000 individual members, generated as combinatorial M13 phage display and synthetic peptide libraries, respectively, with structural composition GWXPXDXPI, where X is any of 20 natural amino acids. Statistically significant tumor growth inhibition was observed in BALB/c mice immunized with the VELs in both prophylactic and therapeutic settings. Vaccinated mice developed epitope-specific spleen cell and CD8+ IFN-γ+ T-cell responses that recognize more than 50% of the panel of 87 mutated epitope variants, as demonstrated in T-cell proliferation assays and FACS analysis. These data indicate the feasibility of the application of this new class of immunogens based on VEL concept as an alternative approach for the development of molecular vaccines against cancer.

  15. Differences in associations between active transportation and built environmental exposures when expressed using different components of individual activity spaces.

    Science.gov (United States)

    van Heeswijck, Torbjorn; Paquet, Catherine; Kestens, Yan; Thierry, Benoit; Morency, Catherine; Daniel, Mark

    2015-05-01

    This study assessed relationships between built environmental exposures measured within components of individual activity spaces (i.e., travel origins, destinations and paths in-between), and use of active transportation in a metropolitan setting. Individuals (n=37,165) were categorised as using active or sedentary transportation based on travel survey data. Generalised Estimating Equations analysis was used to test relationships with active transportation. Strength and significance of relationships between exposures and active transportation varied for different components of the activity space. Associations were strongest when including travel paths in expression of the built environment. Land use mix and greenness were negatively related to active transportation.

  16. SIRT1 expression is associated with poor prognosis of lung adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Li C

    2015-04-01

    Full Text Available Chong Li,1,2,* Lingling Wang,3,* Liang Zheng,4 Xianghong Zhan,4 Bin Xu,1,2 Jingting Jiang,1,2 Changping Wu1,2 1Department of Tumor Biological Treatment, the Third Affiliated Hospital, Soochow University, Changzhou, 2Cancer Immunotherapy Engineering Research Center of Jiangsu Province, Changzhou, 3Department of Medical Education, Jinling Hospital, Medical School of Nanjing University, Nanjing, 4Department of Thoracic Surgery, the Third Affiliated Hospital, Soochow University, Changzhou, Jiangsu, People’s Republic of China *These authors contributed equally to this work Abstract: Several studies have reported that the overexpression of Sirtuin 1 (SIRT1 was associated with poor prognosis in various human cancers. However, little is known regarding the prognostic value of SIRT1 in lung adenocarcinoma. Therefore, the aim of this study is to evaluate the role of SIRT1 in the prognosis of lung adenocarcinoma patients. Using a tissue microarray, we detected SIRT1 expression by immunohistochemistry in lung adenocarcinoma tissue, as well as in corresponding noncancerous tissues (NCTs. A high expression level of SIRT1 was observed in 74.7% (56/75 of patients with lung adenocarcinoma and 6.7% (5/75 of NCTs (P<0.001. SIRT1 expression was significantly associated with high pathological stage. Importantly, we found that SIRT1 expression was associated with worse overall survival in these lung adenocarcinoma patients (67.0 months vs 104.5 months; P=0.005. In addition, anaplastic lymphoma kinase, epidermal growth factor receptor, vascular endothelial growth factor (VEGF, and Survivin expression were evaluated by fluorescent in situ hybridization or immunohistochemistry, respectively. We found that VEGF and Survivin were both highly expressed in the lung adenocarcinoma tissues, as compared to NCTs. Moreover, the SIRT1 and VEGF expression statuses were significantly positively correlated (r=0.238, P=0.039, while SIRT1 and Survivin expression status were not

  17. Reactive oxygen species in signalling the transcriptional activation of WIPK expression in tobacco.

    Science.gov (United States)

    Xu, Juan; Yang, Kwang-Yeol; Yoo, Seung Jin; Liu, Yidong; Ren, Dongtao; Zhang, Shuqun

    2014-07-01

    Plant mitogen-activated protein kinases represented by tobacco WIPK (wounding-induced protein kinase) and its orthologs in other species are unique in their regulation at transcriptional level in response to stress and pathogen infection. We previously demonstrated that transcriptional activation of WIPK is essential for induced WIPK activity, and activation of salicylic acid-induced protein kinase (SIPK) by the constitutively active NtMEK2(DD) is sufficient to induce WIPK gene expression. Here, we report that the effect of SIPK on WIPK gene expression is mediated by reactive oxygen species (ROS). Using a combination of pharmacological and gain-of-function transgenic approaches, we studied the relationship among SIPK activation, WIPK gene activation in response to fungal cryptogein, light-dependent ROS generation in chloroplasts, and ROS generated via NADPH oxidase. In the conditional gain-of-function GVG-NtMEK2(DD) transgenic tobacco, induction of WIPK expression is dependent on the ROS generation in chloroplasts. Consistently, methyl viologen, an inducer of ROS generation in chloroplasts, highly activated WIPK expression. In addition to chloroplast-originated ROS, H(2)O(2) generated from the cell-surface NADPH oxidase could also activate WIPK gene expression, and inhibition of cryptogein-induced ROS generation also abolished WIPK gene activation. Our data demonstrate that WIPK gene activation is mediated by ROS, which provides a mechanism by which ROS influence cellular signalling processes in plant stress/defence response.

  18. Ornithine decarboxylase, mitogen-activated protein kinase and matrix metalloproteinase-2 expressions in human colon tumors

    Institute of Scientific and Technical Information of China (English)

    Takahiro Nemoto; Shunichiro Kubota; Hideyuki Ishida; Nobuo Murata; Daijo Hashimoto

    2005-01-01

    AIM: To investigate the expressions of omithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors.METHODS: ODC activity, MMP-2 expression, and mitogenactivated protein (MAP) kinase activity (Erk phosphorylation) were determined in 58 surgically removed human colon tumors and their adjacent normal tissues, using [1-14C]-ornithine as a substrate, ELISA assay, and Western blotting, respectively.RESULTS: ODC activity, MMP-2 expression, and Erk phosphorylation were significantly elevated in colon tumors, compared to those in adjacent normal tissues. A significant correlation was observed between ODC activities and MMP-2 levels.CONCLUSION: This is the first report showing a significant correlation between ODC activities and MMP-2 levels in human colon tumors. As MMP-2 is involved in cancer invasion and metastasis, and colon cancer overexpresses ODC, suppression of ODC expression may be a rational approach to treat colon cancer which overexpresses ODC.

  19. Survivin反义寡核苷酸协同Taxol诱导肺癌细胞株凋亡%Effect of Survivin Antisense Oligonucleotide Combined with Taxol on Induced Apoptosis in Lung Cancer Cells

    Institute of Scientific and Technical Information of China (English)

    陈余清; 夏雪梅; 蔡映云; 黄礼年; 李殿明; 胡俊锋

    2005-01-01

    [目的]研究Survivin反义寡核苷酸(antisense oligonucleotide,ASODN)单独或联合Taxol对肺癌细胞株Survivin mRNA和蛋白表达,细胞凋亡,生长抑制率的影响.[方法]Survivin ASODN经脂质体介导转染人小细胞肺癌细胞株NCI-H446,用RT-PCR法、Western blot法检测Survivin表达;Survivin ASODN单独、联合Taxol作用NCI-H446细胞后,用MTT法检测细胞生长抑制率,台盼兰拒染实验检测细胞死亡率,流式细胞仪检测细胞凋亡并计算两药相互作用指数(CDI).[结果]Survivin ASODN转染NCI-H446细胞后,Survivin mRNA表达和蛋白表达明显下调,其中SurvivinASODN 500nM作用72h时Survivin mRNA抑制率达62.72%,效果最佳;Survivin ASODN单独或联合Taxol作用NCI-H446细胞后发现Survivin ASODN联合Taxol作用的效果明显优于两药单独应用(P<0.01).其联用时细胞凋亡率达73.3%,而单用时分别为43.6%和23.8%.其联用时细胞生长抑制率达80.1%,而单用时抑制率分别为50.4%和30.5%(P均<0.01).两药联用组细胞死亡率达69.9%,高于两药单用时的41.4%和24.8%(P均<0.01);CDI值为0.43,表明两药具有显著协同作用.[结论]Survivin ASODN能够抑制肺癌细胞株Survivin mRNA和蛋白表达并诱导肺癌细胞凋亡;Survivin ASODN能够增加Taxol的敏感性.

  20. Neuronal MHC Class I Expression Is Regulated by Activity Driven Calcium Signaling.

    Directory of Open Access Journals (Sweden)

    Dan Lv

    Full Text Available MHC class I (MHC-I molecules are important components of the immune system. Recently MHC-I have been reported to also play important roles in brain development and synaptic plasticity. In this study, we examine the molecular mechanism(s underlying activity-dependent MHC-I expression using hippocampal neurons. Here we report that neuronal expression level of MHC-I is dynamically regulated during hippocampal development after birth in vivo. Kainic acid (KA treatment significantly increases the expression of MHC-I in cultured hippocampal neurons in vitro, suggesting that MHC-I expression is regulated by neuronal activity. In addition, KA stimulation decreased the expression of pre- and post-synaptic proteins. This down-regulation is prevented by addition of an MHC-I antibody to KA treated neurons. Further studies demonstrate that calcium-dependent protein kinase C (PKC is important in relaying KA simulation activation signals to up-regulated MHC-I expression. This signaling cascade relies on activation of the MAPK pathway, which leads to increased phosphorylation of CREB and NF-κB p65 while also enhancing the expression of IRF-1. Together, these results suggest that expression of MHC-I in hippocampal neurons is driven by Ca2+ regulated activation of the MAPK signaling transduction cascade.

  1. Role of the STAT3/survivin signaling pathway in the EML4-ALK-positive lung adenocarcinoma cell line H2228 before and after crizotinib-induced resistance

    Institute of Scientific and Technical Information of China (English)

    Haiyan Peng; Wenhua Zhao Co-first author; Cuiyun Su; Xiangqun Song; Aiping Zeng; Huilin Wang; Ruiling Ning; Shaozhang Zhou 

    2015-01-01

    Objective This study investigated the role of the STAT3/survivin signaling pathway in the EML4-ALK–positive lung adenocarcinoma cel line H2228 before and after crizotinib-induced resistance. The mecha-nism of resistance was studied. Methods Cel viability was determined using the MTT assay. Crizotinib-induced apoptosis in H2228 and H2228 crizotinib-resistant cel s treated with the indicated doses of crizotinib was measured at dif erent times (24 h, 48 h, 72 h) using flow cytometry. The levels of p-ALK, ALK, p-STAT3, STAT3, and survivin after treatment of cel s with 0, 0.3, and 1μM crizotinib for 72 h were determined using Western blot analysis. DNA sequencing was used to identify mutations in H2228 crizotinib-resistant cel s. Results The crizotinib IC50 values in H2228 and H2228 crizotinib-resistant cel s at 72 h were 334.5 nM and 3418 nM, respectively. The resistance index of H2228 crizotinib-resistant cel s was 10.20. Crizotinib induced apoptosis in H2228 cel s and reduced the levels of p-ALK, p-STAT3, and survivin. In contrast, no changes in the levels of p-ALK, p-STAT3, and survivin were observed in H2228 crizotinib-resistant cel s. The mutations 2067G→A and 2182G→C in EML4-ALK were present in the H2228 crizotinib-resistant cel s. Conclusion Crizotinib decreased the viability of H2228 cel s in a dose- and time-dependent manner. In the STAT3/survivin pathway, downregulation of p-ALK, p-STAT3, and survivin might contribute to crizo-tinib-induced apoptosis in H2228 cel s. However, the STAT3/survivin pathway in H2228 crizotinib-resistant cel s was unaf ected by crizotinib treatment. Acquired resistance in H2228 cel s might be related to ALK mutations.

  2. Growth enhancement and gene expression of Arabidopsis thaliana irradiated with active oxygen species

    Science.gov (United States)

    Watanabe, Satoshi; Ono, Reoto; Hayashi, Nobuya; Shiratani, Masaharu; Tashiro, Kosuke; Kuhara, Satoru; Inoue, Asami; Yasuda, Kaori; Hagiwara, Hiroko

    2016-07-01

    The characteristics of plant growth enhancement effect and the mechanism of the enhancement induced by plasma irradiation are investigated using various active species in plasma. Active oxygen species in oxygen plasma are effective for growth enhancement of plants. DNA microarray analysis of Arabidopsis thaliana indicates that the genes coding proteins that counter oxidative stresses by eliminating active oxygen species are expressed at significantly high levels. The size of plant cells increases owing to oxygen plasma irradiation. The increases in gene expression levels and cell size suggest that the increase in the expression level of the expansin protein is essential for plant growth enhancement phenomena.

  3. Cytotoxicity of calotropin is through caspase activation and downregulation of anti-apoptotic proteins in K562 cells.

    Science.gov (United States)

    Wang, Shih-Chung; Lu, Mei-Chin; Chen, Hsiu-Lin; Tseng, Hsing-I; Ke, Yu-Yuan; Wu, Yang-Chang; Yang, Pei-Yu

    2009-12-01

    Calotropin is one of cardenolides isolated from milkweed used for medicinal purposes in many Asian countries. Whereas calotropin possesses cytotoxicity against several cancer cells, the mechanisms of action remain unclear. We set out to evaluate the cytotoxic mechanism of calotropin on human chronic myeloid leukemia K562 cells. Calotropin inhibited the growth of K562 cells in a time- and dose-dependent manner by G(2)/M phase arrest. It upregulated the expression of p27 leading to this arrest by downregulating the G2/M regulatory proteins, cyclins A and B, and by upregulating the cdk inhibitor, p27. Furthermore, it downregulated anti-apoptotic signaling (XIAP and survivin) and survival pathways (p-Akt and NFkappaB), leading to caspase-3 activation which resulted in the induction of apoptosis. In all, calotropin exerted its anticancer activity on K562 cells by modulating the pro-survival signaling that leads to induction of apoptosis.

  4. Effects of fulvestrant on biological activity and Wnt expression in rat GH3 cells

    Institute of Scientific and Technical Information of China (English)

    Jiwei Bai; Yan Wang; Chuzhong Li; Yazhuo Zhang

    2012-01-01

    The present study investigated the influence of anti-estrogen treatment (fulvestrant) on pituitary adenoma cell line GH3 biological activity, the estrogen receptor α pathway, the WnT pathway, and mechanisms of decreased Wnt inhibitory factor-1 expression in GH3 cells. Results showed that fulvestrant suppressed GH3 cell proliferation and reduced hormone secretion in a dose-dependent manner. Estrogen receptor α and Wnt4 expression decreased, but Wnt inhibitory factor-1 expression increased in a dose-dependent manner following fulvestrant treatment, and β-catenin expression remained unchanged. Inhibitors of DNA methylation and histone modification upregulated Wnt inhibitory factor-1 expression. Results suggested that fulvestrant suppressed biological activity of GH3 cells via the estrogen receptor α and Wnt pathways. These results suggested that decreased Wnt inhibitory factor-1 expression in GH3 cells played a role in epigenetic mechanisms. Anti-estrogen therapies could provide novel treatments for growth hormone adenomas.

  5. Signal transduction by HLA class II antigens expressed on activated T cells

    DEFF Research Database (Denmark)

    Ødum, Niels; Martin, P J; Schieven, G L;

    1991-01-01

    Human T cells express HLA class II antigens upon activation. Although activated, class II+ T cells can present alloantigens under certain circumstances, the functional role of class II antigens on activated T cells remains largely unknown. Here, we report that cross-linking of HLA-DR molecules ex...

  6. Activation of perineuronal net-expressing excitatory neurons during associative memory encoding and retrieval

    Science.gov (United States)

    Morikawa, Shota; Ikegaya, Yuji; Narita, Minoru; Tamura, Hideki

    2017-01-01

    Perineuronal nets (PNNs), proteoglycan-rich extracellular matrix structures, are thought to be expressed around inhibitory neurons and contribute to critical periods of brain function and synaptic plasticity. However, in some specific brain regions such as the amygdala, PNNs were predominantly expressed around excitatory neurons. These neurons were recruited during auditory fear conditioning and memory retrieval. Indeed, the activation of PNN-expressing excitatory neurons predicted cognitive performance. PMID:28378772

  7. Neuronal Activity Regulates Hippocampal miRNA Expression

    NARCIS (Netherlands)

    Eacker, Stephen M.; Keuss, Matthew J.; Berezikov, Eugene; Dawson, Valina L.; Dawson, Ted M.

    2011-01-01

    Neuronal activity regulates a broad range of processes in the hippocampus, including the precise regulation of translation. Disruptions in proper translational control in the nervous system are associated with a variety of disorders that fall in the autistic spectrum. MicroRNA (miRNA) represent a re

  8. HLA-DR expression and disease activity in ulcerative colitis

    DEFF Research Database (Denmark)

    Poulsen, L O; Elling, P; Sørensen, Flemming Brandt

    1986-01-01

    In 12 patients with active ulcerative colitis (UC) the rectal epithelial cells were analyzed for HLA-DR antigens by an immunohistochemical technique. The clinical, rectoscopic, and histologic stages were also determined. The investigations were carried out at the beginning of the study and 2 weeks...

  9. Expression and Activation of STAT Transcription Factors in Breast Cancer

    Science.gov (United States)

    1998-05-08

    clinicians. J~, 273: 577-585, 1995. 183 Hundertmark 5, Buhler H, Rudolf M, Weitzel HK, Ragosch V: Inhibition of 11 beta-hydroxysteroid dehydrogenase...activated protein kinase through a Jakl-dependent pathway. Mol. Cell. Bioi., 17:3833-40, 1997. Stewart JF, Rubens RO, King RJ, Minton MJ, Steiner R

  10. Telomerase activity, estrogen receptors (α, β), Bcl-2 expression in human breast cancer and treatment response

    Science.gov (United States)

    Murillo-Ortiz, Blanca; Astudillo-De la Vega, Horacio; Castillo-Medina, Sebastian; Malacara, JM; Benitez-Bribiesca, Luis

    2006-01-01

    Background The mechanism for maintaining telomere integrity is controlled by telomerase, a ribonucleoprotein enzyme that specifically restores telomere sequences, lost during replication by means of an intrinsic RNA component as a template for polymerization. Among the telomerase subunits, hTERT (human telomerase reverse transcriptase) is expressed concomitantly with the activation of telomerase. The role of estrogens and their receptors in the transcriptional regulation of hTERT has been demonstrated. The current study determines the possible association between telomerase activity, the expression of both molecular forms of estrogen receptor (ERα and ERβ) and the protein bcl-2, and their relative associations with clinical parameters. Methods Tissue samples from 44 patients with breast cancer were used to assess telomerase activity using the TRAP method and the expression of ERα, ERβ and bcl-2 by means of immunocytochemical techniques. Results Telomerase activity was detected in 59% of the 44 breast tumors examined. Telomerase activity ranged from 0 to 49.93 units of total product generated (TPG). A correlation was found between telomerase activity and differentiation grade (p = 0.03). The only significant independent marker of response to treatment was clinical stage. We found differences between the frequency of expression of ERα (88%) and ERβ (36%) (p = 0.007); bcl-2 was expressed in 79.5% of invasive breast carcinomas. We also found a significant correlation between low levels of telomerase activity and a lack of ERβ expression (p = 0.03). Conclusion Lower telomerase activity was found among tumors that did not express estrogen receptor beta. This is the first published study demonstrating that the absence of expression of ERβ is associated with low levels of telomerase activity. PMID:16911782

  11. Telomerase activity, estrogen receptors (α, β, Bcl-2 expression in human breast cancer and treatment response

    Directory of Open Access Journals (Sweden)

    Malacara JM

    2006-08-01

    Full Text Available Abstract Background The mechanism for maintaining telomere integrity is controlled by telomerase, a ribonucleoprotein enzyme that specifically restores telomere sequences, lost during replication by means of an intrinsic RNA component as a template for polymerization. Among the telomerase subunits, hTERT (human telomerase reverse transcriptase is expressed concomitantly with the activation of telomerase. The role of estrogens and their receptors in the transcriptional regulation of hTERT has been demonstrated. The current study determines the possible association between telomerase activity, the expression of both molecular forms of estrogen receptor (ERα and ERβ and the protein bcl-2, and their relative associations with clinical parameters. Methods Tissue samples from 44 patients with breast cancer were used to assess telomerase activity using the TRAP method and the expression of ERα, ERβ and bcl-2 by means of immunocytochemical techniques. Results Telomerase activity was detected in 59% of the 44 breast tumors examined. Telomerase activity ranged from 0 to 49.93 units of total product generated (TPG. A correlation was found between telomerase activity and differentiation grade (p = 0.03. The only significant independent marker of response to treatment was clinical stage. We found differences between the frequency of expression of ERα (88% and ERβ (36% (p = 0.007; bcl-2 was expressed in 79.5% of invasive breast carcinomas. We also found a significant correlation between low levels of telomerase activity and a lack of ERβ expression (p = 0.03. Conclusion Lower telomerase activity was found among tumors that did not express estrogen receptor beta. This is the first published study demonstrating that the absence of expression of ERβ is associated with low levels of telomerase activity.

  12. The Body Action Coding System II: Muscle activations during the perception and expression of emotion

    Directory of Open Access Journals (Sweden)

    Elisabeth M.J. Huis in 't Veld

    2014-09-01

    Full Text Available Research into the expression and perception of emotions has mostly focused on facial expressions. Recently, body postures have become increasingly important in research, but knowledge on muscle activity during the perception or expression of emotion is lacking. The current study continues the development of a Body Action Coding System (BACS, which was initiated in a previous study, and described the involvement of muscles in the neck, shoulders and arms during expression of fear and anger. The current study expands the BACS by assessing the activity patterns of three additional muscles. Surface electromyography of muscles in the neck (upper trapezius descendens, forearms (extensor carpi ulnaris, lower back (erector spinae longissimus and calves (peroneus longus were measured during active expression and passive viewing of fearful and angry body expressions. The muscles in the forearm were strongly active for anger expression and to a lesser extent for fear expression. In contrast, muscles in the calves were recruited slightly more for fearful expressions. It was also found that muscles automatically responded to the perception of emotion, without any overt movement. The observer’s forearms responded to the perception of fear, while the muscles used for leaning backwards were activated when faced with an angry adversary. Lastly, the calf responded immediately when a fearful person was seen, but responded slower to anger. There is increasing interest in developing systems that are able to create or recognize emotional body language for the development of avatars, robots, and online environments. To that end, multiple coding systems have been developed that can either interpret or create bodily expressions based on static postures, motion capture data or videos. However, the BACS is the first coding system based on muscle activity.

  13. The Body Action Coding System II: muscle activations during the perception and expression of emotion.

    Science.gov (United States)

    Huis In 't Veld, Elisabeth M J; van Boxtel, Geert J M; de Gelder, Beatrice

    2014-01-01

    Research into the expression and perception of emotions has mostly focused on facial expressions. Recently, body postures have become increasingly important in research, but knowledge on muscle activity during the perception or expression of emotion is lacking. The current study continues the development of a Body Action Coding System (BACS), which was initiated in a previous study, and described the involvement of muscles in the neck, shoulders and arms during expression of fear and anger. The current study expands the BACS by assessing the activity patterns of three additional muscles. Surface electromyography of muscles in the neck (upper trapezius descendens), forearms (extensor carpi ulnaris), lower back (erector spinae longissimus) and calves (peroneus longus) were measured during active expression and passive viewing of fearful and angry body expressions. The muscles in the forearm were strongly active for anger expression and to a lesser extent for fear expression. In contrast, muscles in the calves were recruited slightly more for fearful expressions. It was also found that muscles automatically responded to the perception of emotion, without any overt movement. The observer's forearms responded to the perception of fear, while the muscles used for leaning backwards were activated when faced with an angry adversary. Lastly, the calf responded immediately when a fearful person was seen, but responded slower to anger. There is increasing interest in developing systems that are able to create or recognize emotional body language for the development of avatars, robots, and online environments. To that end, multiple coding systems have been developed that can either interpret or create bodily expressions based on static postures, motion capture data or videos. However, the BACS is the first coding system based on muscle activity.

  14. Gene expression in IFN-g-activated murine macrophages

    Directory of Open Access Journals (Sweden)

    Pereira C.A.

    2004-01-01

    Full Text Available Macrophages are critical for natural immunity and play a central role in specific acquired immunity. The IFN-gamma activation of macrophages derived from A/J or BALB/c mice yielded two different patterns of antiviral state in murine hepatitis virus 3 infection, which were related to a down-regulation of the main virus receptor. Using cDNA hybridization to evaluate mRNA accumulation in the cells, we were able to identify several genes that are differently up- or down-regulated by IFN-gamma in A/J (267 and 266 genes, respectively, up- and down-regulated or BALB/c (297 and 58 genes, respectively, up- and down-regulated mouse macrophages. Macrophages from mice with different genetic backgrounds behave differently at the molecular level and comparison of the patterns of non-activated and IFN-gamma-activated A/J or BALB/c mouse macrophages revealed, for instance, an up-regulation and a down-regulation of genes coding for biological functions such as enzymatic reactions, nucleic acid synthesis and transport, protein synthesis, transport and metabolism, cytoskeleton arrangement and extracellular matrix, phagocytosis, resistance and susceptibility to infection and tumors, inflammation, and cell differentiation or activation. The present data are reported in order to facilitate future correlation of proteomic/transcriptomic findings as well as of results obtained from a classical approach for the understanding of biological phenomena. The possible implication of the role of some of the gene products relevant to macrophage biology can now be further scrutinized. In this respect, a down-regulation of the main murine hepatitis virus 3 receptor gene was detected only in IFN-gamma-activated macrophages of resistant mice.

  15. Indoleamine 2,3 Dioxygenase (IDO Expression and Activity in Relapsing-Remitting Multiple Sclerosis.

    Directory of Open Access Journals (Sweden)

    Roberta Mancuso

    Full Text Available Interferon gamma (IFN-γ production induces the transcription of indoleamine 2,3 dioxygenase (IDO resulting in the reduction of T-cell activation and proliferation through the depletion of tryptophan and the elicitation of Treg lymphocytes. IDO was shown to be involved in the pathogenesis of autoimmune diseases; we investigated whether changes in IDO gene expression and activity could be indicative of onset of relapse in multiple sclerosis (MS patients.IDO and interferon-γ (IFN-γ gene expression, serum IDO activity (Kynurenine/Tryptophan ratio and serum neopterin concentration--a protein released by macrophages upon IFN-γ stimulation--were measured in 51 individuals: 36 relapsing remitting (RR-MS patients (21 in acute phase--AMS, 15 in stable phase--SMS and 15 healthy controls (HC. PBMCs samples in AMS patients were collected before (BT-AMS and during glucocorticoids-based therapy (DT-AMS.IDO expression was increased and IFN-γ was decreased (p<0.001 in BT-AMS compared to SMS patients. Glucocorticoids-induced disease remission resulted in a significant reduction of IDO and IFN-γ gene expression, IDO catalytic activity (p<0.001. Serum neopterin concentration followed the same trend as IDO expression and activity.Measurement of IDO gene expression and activity in blood could be a useful marker to monitor the clinical course of RR-MS. Therapeutic interventions modulating IDO activity may be beneficial in MS.

  16. TELOMERASE ACTIVITY IN COLORECTAL CARCINOMA AND ITS CORRELATION WITH EXPRESSION OF C-MYC

    Institute of Scientific and Technical Information of China (English)

    LIU Jian-Lun; GE Lian-ying; ZHANG Gui-nian

    2005-01-01

    Objective: To study the role of telomerase activity and c-myc in pathogenesis and progression of colorectal carcinoma,and to investigate the possible regulatory mechanism of telomerase activation. Methods: A modified telomeric repeat amplification protocol (TRAP) and immunohistochemical staining was used to detect telomerase activity and the expression of c-myc in tissue samples from colorectal carcinoma, paracarcinomatousl tissues, normal mucosa, and adenomatoid polyp.Results: The positive rates of telomerase activity and c-myc expression were 83.33% and 80.00% in colorectal carcinoma,13.33% and 23.33% in paracarcinomatousl tissues, 13.33% and 20.00% in normal mucosa, and 10.00% and 45.00% in adenomatoid polyp respectively, they were significantly higher in colorectal carcinoma than in paracarcinomatousl tissues,normal mucosa, and adenomatoid polyp (P<0.05). The rates of telomerase activity and c-myc expression were much higher in colorectal carcinoma with lymph nodes metastases than that without lymph nodes metastases. The expression of c-myc was found being significantly higher in the telomerase positive colorectal carcinoma than in the telomerase negative group(P<0.05). Conclusion: The activation of telomerase and abnormal expression of c-myc might play an important role in the process of carcinogenesis and progression of colorectal carcinoma. The over-expression of c-myc may be related to telomerase activation and up-regulation in colorectal carcinoma.

  17. Decreased Expression of SRSF2 Splicing Factor Inhibits Apoptotic Pathways in Renal Cancer

    Science.gov (United States)

    Kędzierska, Hanna; Popławski, Piotr; Hoser, Grażyna; Rybicka, Beata; Rodzik, Katarzyna; Sokół, Elżbieta; Bogusławska, Joanna; Tański, Zbigniew; Fogtman, Anna; Koblowska, Marta; Piekiełko-Witkowska, Agnieszka

    2016-01-01

    Serine and arginine rich splicing factor 2(SRSF2) belongs to the serine/arginine (SR)-rich family of proteins that regulate alternative splicing. Previous studies suggested that SRSF2 can contribute to carcinogenic processes. Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer, highly aggressive and difficult to treat, mainly due to resistance to apoptosis. In this study we hypothesized that SRSF2 contributes to the regulation of apoptosis in ccRCC. Using tissue samples obtained from ccRCC patients, as well as independent validation on The Cancer Genome Atlas (TCGA) data, we demonstrate for the first time that expression of SRSF2 is decreased in ccRCC tumours when compared to non-tumorous control tissues. Furthermore, by employing a panel of ccRCC-derived cell lines with silenced SRSF2 expression and qPCR arrays we show that SRSF2 contributes not only to splicing patterns but also to expression of multiple apoptotic genes, including new SRSF2 targets: DIABLO, BIRC5/survivin, TRAIL, BIM, MCL1, TNFRSF9, TNFRSF1B, CRADD, BCL2L2, BCL2A1, and TP53. We also identified a new splice variant of CFLAR, an inhibitor of caspase activity. These changes culminate in diminished caspase-9 activity and inhibition of apoptosis. In summary, we show for the first time that decreased expression of SRSF2 in ccRCC contributes to protection of cancer cells viability. PMID:27690003

  18. Activation of hepatic lipase expression by oleic acid: possible involvement of USF1.

    NARCIS (Netherlands)

    D. van Deursen (Diederik); M. van Leeuwen (Marije); D. Akdogan (Deniz); H. Adams (Hadie); H. Jansen (Hans); A.J.M. Verhoeven (Adrie)

    2009-01-01

    textabstractPolyunsaturated fatty acids affect gene expression mainly through peroxisome proliferator-activated receptors (PPARs) and sterol regulatory element binding proteins (SREBPs), but how monounsaturated fatty acids affect gene expression is poorly understood. In HepG2 cells, oleate supplemen

  19. Cyclooxygenase-2 expression is dependent upon epidermal growth factor receptor expression or activation in androgen independent prostate cancer

    Institute of Scientific and Technical Information of China (English)

    Rui-Peng Jia; Lu-Wei Xu; Qi Su; Jian-Hua Zhao; Wen-Cheng Li; Feng Wang; Zheng Xu

    2008-01-01

    Aim: To investigate the expression of cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) and the possible mechanism in the development in androgen independent prostate cancer (AIPC). Methods: Immunohis- tochemistry was performed on paraffin-embedded sections with goat polyclonal against COX-2 and mouse mono- clonal antibody against EGFR in 30 AIPC and 18 androgen dependent prostate cancer (ADPC) specimens. The effect of epidermal growth factor (EGF) treatments on the expression of COX-2 and signal pathway in PC-3 and DU-145 cells was studied using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. ELISA was used to measure prostaglandin E2 (PGE2) levels in the media of PC-3 and DU-145 incubated with EGF for 24 h. Results: COX-2 was positively expressed in AIPC and ADPC, which were predominantly in endochylema of prostate cancer (Pca) cells. Intense staining was seen in AIPC (80%) and in ADPC (55.5%), but there was no significant association between the two groups. EGFR expression was also positive in the two groups (61.8% in ADPC and 90% in AIPC, P < 0.01). A significant association was found between EGFR expression and a higher Gleason score (P < 0.05) or tumor stage (P < 0.05). The expression of PGE2 was increased in PC-3 and DU-145 cells after being incubated with EGF. Both p38MAPK and PI-3K pathway were involved in the PC-3 cell COX-2 upregulation course. In DU- 145, only p38MAPK pathway was associated with COX-2 upregulation. Conclusion: EGFR activation induces COX-2 expression through PI-3K and/or p38MAPK pathways. COX-2 and EGFR inhibitors might have a cooperative anti-tumor effect in Pca.

  20. IL-35 over-expression increases apoptosis sensitivity and suppresses cell growth in human cancer cells.

    Science.gov (United States)

    Long, Jun; Zhang, Xulong; Wen, Mingjie; Kong, Qingli; Lv, Zhe; An, Yunqing; Wei, Xiao-Qing

    2013-01-01

    Interleukin (IL)-35 is a novel heterodimeric cytokine in the IL-12 family and is composed of two subunits: Epstein-Barr virus-induced gene 3 (EBI3) and IL-12p35. IL-35 is expressed in T regulatory (Treg) cells and contributes to the immune suppression function of these cells. In contrast, we found that both IL-35 subunits were expressed concurrently in most human cancer cell lines compared to normal cell lines. In addition, we found that TNF-α and IFN-γ stimulation led to increased IL-35 expression in human cancer cells. Furthermore, over-expression of IL-35 in human cancer cells suppressed cell growth in vitro, induced cell cycle arrest at the G1 phase, and mediated robust apoptosis induced by serum starvation, TNF-α, and IFN-γ stimulation through the up-regulation of Fas and concurrent down-regulation of cyclinD1, survivin, and Bcl-2 expression. In conclusion, our results reveal a novel functional role for IL-35 in suppressing cancer activity, inhibiting cancer cell growth, and increasing the apoptosis sensitivity of human cancer cells through the regulation of genes related to the cell cycle and apoptosis. Thus, this research provides new insights into IL-35 function and presents a possible target for the development of novel cancer therapies.

  1. Expression of activated Ras during Dictyostelium development alters cell localization and changes cell fate.

    Science.gov (United States)

    Jaffer, Z M; Khosla, M; Spiegelman, G B; Weeks, G

    2001-03-01

    There is now a body of evidence to indicate that Ras proteins play important roles in development. Dictyostelium expresses several ras genes and each appears to perform a distinct function. Previous data had indicated that the overexpression of an activated form of the major developmentally regulated gene, rasD, caused a major aberration in morphogenesis and cell type determination. We now show that the developmental expression of an activated rasG gene under the control of the rasD promoter causes a similar defect. Our results indicate that the expression of activated rasG in prespore cells results in their transdifferentiation into prestalk cells, whereas activated rasG expression in prestalk causes gross mislocalization of the prestalk cell populations.

  2. Ly-6A is required for T cell receptor expression and protein tyrosine kinase fyn activity.

    Science.gov (United States)

    Lee, S K; Su, B; Maher, S E; Bothwell, A L

    1994-05-01

    To characterize the function of the Ly-6A antigen in T cell activation, antisense Ly-6 RNA was expressed in a stably transfected antigen-specific T cell clone. Reduced Ly-6A expression results in inhibition of responses to antigen, anti-TCR (anti-T cell receptor) crosslinking and concanavalin A plus recombinant interleukin 1 and causes impairment of in vitro fyn tyrosine kinase activity. More substantial reduction of Ly-6A results in reduction of TCR expression. Analysis of mRNA species indicates that the reduction is specific for the TCR beta chain. These data demonstrate that Ly-6A may regulate TCR expression and may be involved in early events of T cell activation via regulation of fyn tyrosine kinase activity.

  3. Spontaneous and cytokine induced expression and activity of matrix metalloproteinases in human colonic epithelium

    DEFF Research Database (Denmark)

    Pedersen, G; Saermark, T; Kirkegaard, T;

    2009-01-01

    for MMP cleavage. HT-29 and DLD-1 expressed several MMPs and levels of MMP-3, -10 and -13 mRNA expression were increased significantly by tumour necrosis factor (TNF)-alpha exposure. Transcripts of MMP-1, -3, -7, -9, -10 and -12 were detected in CECs and all, except MMP12, at significantly increased...... levels in cells from inflamed IBD mucosa. MMP-2 and -8 mRNA were expressed inconsistently and MMP-11, -13 and -14 mRNA undetectable. Proteolytic MMP activity was detected in CEC supernatants and the level was increased significantly in inflamed IBD epithelium. The enzyme activity was inhibited strongly...... by a specific MMP inhibitor (GM 6001). A significant TNF-alpha-mediated increase in MMP enzyme activity was also detected in HT-29 cells in vitro. In conclusion, the expression of several MMPs as well as the level of functional MMPactivity is increased in CEC from patients with active IBD. The results suggest...

  4. Building predictive gene signatures through simultaneous assessment of transcription factor activation and gene expression.

    Science.gov (United States)

    Building predictive gene signatures through simultaneous assessment of transcription factor activation and gene expression Exposure to many drugs and environmentally-relevant chemicals can cause adverse outcomes. These adverse outcomes, such as cancer, have been linked to mol...

  5. Transgenic chickens expressing human urokinase-type plasminogen activator.

    Science.gov (United States)

    Lee, Sung Ho; Gupta, Mukesh Kumar; Ho, Young Tae; Kim, Teoan; Lee, Hoon Taek

    2013-09-01

    Urokinase-type plasminogen activator is a serine protease that is clinically used in humans for the treatment of thrombolytic disorders and vascular diseases such as acute ischemic stroke and acute peripheral arterial occlusion. This study explored the feasibility of using chickens as a bioreactor for producing human urokinase-type plasminogen activator (huPA). Recombinant huPA gene, under the control of a ubiquitous Rous sarcoma virus promoter, was injected into the subgerminal cavity of freshly laid chicken eggs at stage X using the replication-defective Moloney murine leukemia virus (MoMLV)-based retrovirus vectors encapsidated with VSV-G (vesicular stomatitis virus G) glycoprotein. A total of 38 chicks, out of 573 virus-injected eggs, hatched and contained the huPA gene in their various body parts. The mRNA transcript of the huPA gene was present in various organs, including blood and egg, and was germ-line transmitted to the next generation. The level of active huPA protein was 16-fold higher in the blood of the transgenic chicken than in the nontransgenic chicken (P pharming of the huPA drug but also be useful for studying huPA-induced bleeding and other disorders.

  6. Exploring Metrics to Express Energy Expenditure of Physical Activity in Youth

    OpenAIRE

    McMurray, Robert G.; Butte, Nancy F; Scott E Crouter; Trost, Stewart G.; Pfeiffer, Karin A.; Bassett, David R.; Puyau, Maurice R; David Berrigan; Watson, Kathleen B; Fulton, Janet E.

    2015-01-01

    Background Several approaches have been used to express energy expenditure in youth, but no consensus exists as to which best normalizes data for the wide range of ages and body sizes across a range of physical activities. This study examined several common metrics for expressing energy expenditure to determine whether one metric can be used for all healthy children. Such a metric could improve our ability to further advance the Compendium of Physical Activities for Youth. Methods A secondary...

  7. Antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus

    OpenAIRE

    Grabova A. Yu.; Dragovoz I. V.; Zelena L. B.; Tkachuk D. M.; Avdeeva L. V.

    2016-01-01

    Aim. To research the antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus. Methods. Deferred antagonism method, PCR, qRT-PCR, MALDI-TOF mass spectrometry. Results. It was revealed that Bacillus sp. strains C6 and Lg37s out of five tested strains had the highest antifungal activity. Based on the molecular genetic methods, it was shown that the expression of genes of lipopeptide antibiotics, related to the fengycin family, occurred in all these strains...

  8. Stiffness-activated GEF-H1 expression exacerbates LPS-induced lung inflammation.

    Directory of Open Access Journals (Sweden)

    Isa Mambetsariev

    Full Text Available Acute lung injury (ALI is accompanied by decreased lung compliance. However, a role of tissue mechanics in modulation of inflammation remains unclear. We hypothesized that bacterial lipopolysacharide (LPS stimulates extracellular matrix (ECM production and vascular stiffening leading to stiffness-dependent exacerbation of endothelial cell (EC inflammatory activation and lung barrier dysfunction. Expression of GEF-H1, ICAM-1, VCAM-1, ECM proteins fibronectin and collagen, lysyl oxidase (LOX activity, interleukin-8 and activation of Rho signaling were analyzed in lung samples and pulmonary EC grown on soft (1.5 or 2.8 kPa and stiff (40 kPa substrates. LPS induced EC inflammatory activation accompanied by expression of ECM proteins, increase in LOX activity, and activation of Rho signaling. These effects were augmented in EC grown on stiff substrate. Stiffness-dependent enhancement of inflammation was associated with increased expression of Rho activator, GEF-H1. Inhibition of ECM crosslinking and stiffening by LOX suppression reduced EC inflammatory activation and GEF-H1 expression in response to LPS. In vivo, LOX inhibition attenuated LPS-induced expression of GEF-H1 and lung dysfunction. These findings present a novel mechanism of stiffness-dependent exacerbation of vascular inflammation and escalation of ALI via stimulation of GEF-H1-Rho pathway. This pathway represents a fundamental mechanism of positive feedback regulation of inflammation.

  9. Efficient LEC2 activation of OLEOSIN expression requires two neighboring RY elements on its promoter

    Institute of Scientific and Technical Information of China (English)

    CHE NanYing; YANG Yang; LI YanDong; WANG LiLi; HUANG Ping; GAO Yin; An ChengCai

    2009-01-01

    As the main structural protein of oil body, OLEOSIN is highly expressed only during seed development. OLEOSIN promoter is a very useful tool for seed-specific gene engineering and seed bioreactor designing. The B3 domain transcription factor leafy cotyledon2 (LEC2) plays an important role in regulating seed development and seed-specific gene expression. Here, we first report how seed-specific B3 domain transcription factor leafy cotyledon2 (LEC2) efficiently activates OLEOSIN expression. The central promoter region of OLEOSIN, responsible for seed specificity and LEC2 activation, was determined by 5'-deletion analysis. Binding experiments in yeast cells and electrophoretic mobility shift assays showed that LEC2 specifically bound to two conserved RY elements in this region, in transient expression assays, mutation in either RY element dramatically reduced LEC2 activation of OLEOSIN promoter activity, while double mutation abolished it. Analysis of the distribution of RY elements in seed-specific genes activated by LEC2 also supported the idea that genes containing neighboring RY elements responded strongly to LEC2 activation. Therefore, we conclude that two neighboring RY elements are essential for efficient LEC2 activation of OLEOSIN expression. These findings will help us better utilize seed-specific promoter activity.

  10. Efficient LEC2 activation of OLEOSIN expression requires two neighboring RY elements on its promoter

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    As the main structural protein of oil body,OLEOSIN is highly expressed only during seed development. OLEOSIN promoter is a very useful tool for seed-specific gene engineering and seed bioreactor designing. The B3 domain transcription factor leafy cotyledon2 (LEC2) plays an important role in regulating seed development and seed-specific gene expression. Here,we first report how seed-specific B3 domain transcription factor leafy cotyledon2 (LEC2) efficiently activates OLEOSIN expression. The central promoter region of OLEOSIN,responsible for seed specificity and LEC2 activation,was determined by 5’-deletion analysis. Binding experiments in yeast cells and electrophoretic mobility shift assays showed that LEC2 specifically bound to two conserved RY elements in this region. In transient expression assays,mutation in either RY element dramatically reduced LEC2 activation of OLEOSIN promoter activity,while double mutation abolished it. Analysis of the distribution of RY elements in seed-specific genes activated by LEC2 also supported the idea that genes containing neighboring RY elements responded strongly to LEC2 activation. Therefore,we conclude that two neighboring RY elements are essential for efficient LEC2 activation of OLEOSIN expression. These findings will help us better utilize seed-specific promoter activity.

  11. Peroxisome proliferator-activated receptor-gamma suppresses cyclooxygenase-2 expression in human prostate cells.

    Science.gov (United States)

    Sabichi, Anita L; Subbarayan, Vemparala; Llansa, Norma; Lippman, Scott M; Menter, David G

    2004-11-01

    Recent studies have found that cyclooxygenase-2 (COX-2) protein expression was low and inducible with cytokines in prostate cancer cells (in the absence of serum) and that, in contrast, COX-2 expression was high in normal prostate epithelial cells (EC). Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) was expressed at high levels in the prostate cancer cell line PC-3 but not in ECs. In contrast to previous findings by others, PPAR-gamma ligands did not induce PPAR-gamma expression in EC or PC-3. The present study examined the relationship between PPAR-gamma and COX-2 expression patterns in EC and PC-3 in the presence and absence of serum and/or the PPAR-gamma agonist 15-deoxy-Delta12,14-prostaglandin J(2) (15d-PGJ(2)). We also evaluated the effects that the forced expression of PPAR-gamma1 and PPAR-gamma2 had on COX-2 in ECs. We found that expression of PPAR-gamma and COX-2 protein was inversely correlated in ECs and PC-3. Low COX-2 expression in PC-3 was up-regulated by serum, and 15d-PGJ(2) blocked serum-induced COX-2 expression and activity in a dose-dependent manner. 15d-PGJ(2) had no effect on COX-2 expression in ECs or PPAR-gamma expression in either cell type. However, forced expression of PPAR-gamma1 or PPAR-gamma2 in ECs suppressed the high level of endogenous COX-2. This effect was not isoform specific and was augmented by 15d-PGJ(2). The present study showed that PPAR-gamma activation can be an important regulator of COX-2 in prostate cells and may be an important target for prostate cancer chemoprevention.

  12. Relationship between Dyskerin Expression and Telomerase Activity in Human Breast Cancer

    Directory of Open Access Journals (Sweden)

    Lorenzo Montanaro

    2008-01-01

    Full Text Available The nucleolar protein dyskerin is involved in the modification of specific uridine residues to pseudouridine on ribosomal and small nuclear RNAs and in the stabilization of the telomerase RNA component (TERC. In this study we investigated for the first time the relationship between dyskerin expression and telomerase activity in a series of 61 primary breast carcinomas. We found that when dyskerin mRNA values were very low the telomerase activity was markedly reduced, independently of the expression of other important components of the telomerase complex such as telomerase reverse transcriptase (TERT. In vitro experiments showed that reduction of dyskerin expression affect telomerase activity through the reduction of TERC. Only when TERC levels were strongly reduced telomerase activity was hindered. Retroviral mediated over-expression of TERC abolished the telomerase impairment due to dyskerin knock down. In conclusion, our results indicated that, beside its effect on ribosome biogenesis, the levels of dyskerin in cancer cells modulate telomerase activity through the regulation of TERC levels, independently of TERT expression. This should be taken into consideration when utilizing TERT expression as a surrogate indicator of telomerase activity in tumour pathology.

  13. Expression and activities of three inducible enzymes in the healing of gastric ulcers in rats

    Institute of Scientific and Technical Information of China (English)

    Jin-Sheng Guo; Chi-Hin Cho; Wei-Ping Wang; Xi-Zhong Shen; Chuen-Lung Cheng; Marcel Wing Leung Koo

    2003-01-01

    AIM: To explore the roles of nitric oxide synthase (NOS),heme oxygenase (HO) and cyclooxygenase (COX) in gastric ulceration and to investigate the relationships of the expression and activities of these enzymes at different stages of gastric ulceration.METHODS: Gastric ulcers (kissing ulcers) were induced by luminal application of acetic acid. Gastric tissue samples were obtained from the ulcer base, ulcer margin, and nonulcerated area around the ulcer margin at different time intervals after ulcer induction. The mRNA expression and protein levels of inducible and constitutive isoforms of NOS,HO and COX were analyzed with RT-PCR and Western blotting methods. The activities of the total NOS, inducible NOS (iNOS), HO, and COX were also determined.RESULTS: Differential expression of inducible iNOS, HO-1and COX-2 and enzyme activities of NOS, HO and COX were found in the gastric ulcer base. High iNOS expression and activity were observed on day 1 to day 3 in severely inflamed ulcer tissues. Maximum expressions of HO-1 and COX-2 and enzyme activities of HO and COX lagged behind that of iNOS,and remained at high levels during the healing phase.CONCLUSION: The expression and activities of inducible NOS, HO-1 and COX-2 are found to be correlated to different stages of gastric ulceration. Inducible NOS may contribute to ulcer formation while HO-1 and COX-2 may promote ulcer healing.

  14. Telomerase activity and human telomerase reverse transcriptase expression in colorectal carcinoma

    Institute of Scientific and Technical Information of China (English)

    Jian-Lun Liu; Lian-Ying Ge; Gui-Nian Zhang

    2006-01-01

    AIM: To study the activity of telomerase and the expression of human telomerase reverse transcriptase(hTERT) in colorectal carcinoma and its adjacent tissues,normal mucosa and adenomatoid polyp, and to evaluate their relation with carcinogenesis and progression of colorectal carcinoma.METHODS: Telomerase activity and hTERT expression were determined in 30 samples of colorectal carcinoma and its adjacent tissues, normal mucosa and 20samples of adenomatoid polyp by modified telomeric repeat amplification protocol (TRAP), enzyme-linked immunosorbent assay (ELISA) and immunohistochemical method.RESULTS: Telomerase activity and hTERT expression were 83.33% (25/30) and 76.67% (23/30) respectively in colorectal carcinoma, which were obviously higher than those in paracancerous tissues (13.33%, 16.67%),normal mucosa (3.33%, 3.33%) and adenomatoid polyp(10%, 10%). There was a significant difference between colorectal carcinoma and other tissues (P=0.027). The telomerase activity and hTERT expression were higher in colorectal carcinoma with lymphatic metastasis than in that without lymphatic metastasis (P=0.034). When the histological classification and clinical stage were greater,the telomerase activity and hTERT expression increased,but there was no significant difference between them.In colorectal carcinoma, the telomerase activity was correlated with hTERT expression (positive vs negative expression of telomerase activity and hTERT, P=0.021).CONCLUSION: Telomerase activity is closely correlated with the occurrence, development and metastasis of colorectal carcinoma. Overexpression of hTERT may play a critical role in the regulation of telomerase activity.

  15. Influenza matrix protein 2 alters CFTR expression and function through its ion channel activity.

    Science.gov (United States)

    Londino, James D; Lazrak, Ahmed; Jurkuvenaite, Asta; Collawn, James F; Noah, James W; Matalon, Sadis

    2013-05-01

    The human cystic fibrosis transmembrane conductance regulator (CFTR) is a cyclic AMP-activated chloride (Cl(-)) channel in the lung epithelium that helps regulate the thickness and composition of the lung epithelial lining fluid. We investigated whether influenza M2 protein, a pH-activated proton (H(+)) channel that traffics to the plasma membrane of infected cells, altered CFTR expression and function. M2 decreased CFTR activity in 1) Xenopus oocytes injected with human CFTR, 2) epithelial cells (HEK-293) stably transfected with CFTR, and 3) human bronchial epithelial cells (16HBE14o-) expressing native CFTR. This inhibition was partially reversed by an inhibitor of the ubiquitin-activating enzyme E1. Next we investigated whether the M2 inhibition of CFTR activity was due to an increase of secretory organelle pH by M2. Incubation of Xenopus oocytes expressing CFTR with ammonium chloride or concanamycin A, two agents that alkalinize the secretory pathway, inhibited CFTR activity in a dose-dependent manner. Treatment of M2- and CFTR-expressing oocytes with the M2 ion channel inhibitor amantadine prevented the loss in CFTR expression and activity; in addition, M2 mutants, lacking the ability to transport H(+), did not alter CFTR activity in Xenopus oocytes and HEK cells. Expression of an M2 mutant retained in the endoplasmic reticulum also failed to alter CFTR activity. In summary, our data show that M2 decreases CFTR activity by increasing secretory organelle pH, which targets CFTR for destruction by the ubiquitin system. Alteration of CFTR activity has important consequences for fluid regulation and may potentially modify the immune response to viral infection.

  16. Regulation of Neuronal Gene Expression and Survival by Basal NMDA Receptor Activity: A Role for Histone Deacetylase 4

    OpenAIRE

    Chen, Yelin; Wang, Yuanyuan; Modrusan, Zora; Sheng, Morgan; Kaminker, Joshua S.

    2014-01-01

    Neuronal gene expression is modulated by activity via calcium-permeable receptors such as NMDA receptors (NMDARs). While gene expression changes downstream of evoked NMDAR activity have been well studied, much less is known about gene expression changes that occur under conditions of basal neuronal activity. In mouse dissociated hippocampal neuronal cultures, we found that a broad NMDAR antagonist, AP5, induced robust gene expression changes under basal activity, but subtype-specific antagoni...

  17. GILT expression in B cells diminishes cathepsin S steady-state protein expression and activity

    OpenAIRE

    Phipps-Yonas, Hannah; Semik, Vikki; Hastings, Karen Taraszka

    2012-01-01

    MHC class II-restricted Ag processing requires protein degradation in the endocytic pathway for the activation of CD4+ T cells. Gamma-interferon-inducible lysosomal thiol reductase (GILT) facilitates Ag processing by reducing protein disulfide bonds in this compartment. Lysosomal cysteine protease cathepsin S (CatS) contains disulfide bonds and mediates essential steps in MHC class II-restricted processing, including proteolysis of large polypeptides and cleavage of the invariant chain. We so...

  18. Constitutive expression and activity of cytochrome P450 in conventional pigs.

    Science.gov (United States)

    Nielsen, Søren Drud; Bauhaus, Yvonne; Zamaratskaia, Galia; Junqueira, Matheus Antunes; Blaabjerg, Karoline; Petrat-Melin, Bjørn; Young, Jette Feveile; Rasmussen, Martin Krøyer

    2017-04-01

    Pigs have often been suggested to be a useful model for humans, when investigating CYP dependent events, like drug metabolism. However, comprehensive knowledge about the constitutive expression of the major CYP and corresponding transcription factors is limited. We compared the constitutive mRNA expression of aryl hydrocarbon receptor, constitutive androstane receptor and pregnane X receptor and CYP1A1, CYP1A2, CYP2A, CYP2E1 and CYP3A in liver, adipose tissue, muscle and small intestine in pigs, as well as the expression along the length of the small intestine and colon. Tissue samples were taken from female pigs, and analyzed for gene expression, as well as CYP dependent activity using qPCR and specific probe substrates, respectively. For all investigated transcription factors and CYPs the mRNA expression and activity was highest in the liver. CYP1A1 and CYP3A mRNA expression and activity was shown in all investigated tissues. Along the small intestine and colon the mRNA expression and activity of CYP1A1 and CYP3A was gradually decreased. The results demonstrated, similarity to that reported for humans, and hence adds to the use of pigs as a model for humans.

  19. Activation of Hepatic Lipase Expression by Oleic Acid: Possible Involvement of USF1

    Directory of Open Access Journals (Sweden)

    Adrie J. M. Verhoeven

    2009-10-01

    Full Text Available Polyunsaturated fatty acids affect gene expression mainly through peroxisome proliferator-activated receptors (PPARs and sterol regulatory element binding proteins (SREBPs, but how monounsaturated fatty acids affect gene expression is poorly understood. In HepG2 cells, oleate supplementation has been shown to increase secretion of hepatic lipase (HL. We hypothesized that oleate affects HL gene expression at the transcriptional level. To test this, we studied the effect of oleate on HL promoter activity using HepG2 cells and the proximal HL promoter region (700 bp. Oleate increased HL expression and promoter activity 1.3–2.1 fold and reduced SREBP activity by 50%. Downregulation of SREBP activity by incubation with cholesterol+25-hydroxycholesterol had no effect on HL promoter activity. Overexpression of SREBP2, but not SREBP1, reduced HL promoter activity, which was effected mainly through the USF1 binding site at -307/-312. Oleate increased the nuclear abundance of USF1 protein 2.7 ± 0.6 fold, while USF1 levels were reduced by SREBP2 overexpression. We conclude that oleate increases HL gene expression via USF1. USF1 may be an additional fatty acid sensor in liver cells.

  20. Downregulating activated epidermal growth factor receptor has no effect on RBM5 expression

    Institute of Scientific and Technical Information of China (English)

    Twinkle J. Masilamani; Nina D. Rintala-Maki; WANG Ke; Leslie C. Sutherland

    2012-01-01

    Background We were interested in determining how the tumor suppressor gene RBM5 is regulated in lung cancers.Previous studies suggested that the gene expression is related to histological subtype and smoking exposure,since in small cell lung cancers the RBM5 gene is deleted whereas in non-small cell lung carcinomas (NSCLC) RBM5 expression is reduced.Of particular interest was the recent finding that in lung adenocarcinomas,a histological subtype of NSCLC,smoking exposure correlated with mutational activity in the transforming growth factor alpha (TGF-α) signaling pathway.Lung adenocarcinomas from smokers were associated with activating KRAS mutations,whereas lung adenocarcinomas from never-smokers were associated with activating epidermal growth factor receptor (EGFR) mutations.We hypothesized that inhibition of RBM5 in lung adenocarcinomas is achieved indirectly via these activating mutations.The objective of the research described herein was to determine if EGFR activation and RBM5 expression are negatively correlated.Methods EGFR expression in the lung adenocarcinoma cell line NCI-H1975 was inhibited using small interfering RNA.RBM5 expression was examined by real-time quantitative polymerase chain reaction and Western blotting.Results Reduced EGFR expression did not correlate with any change in RBM5 expression at either the RNA or protein level.Conclusion These results suggest that RBM5 expression is not directly regulated by EGFR in non-smoker related lung adenocarinomas,and that some other mechanism operates to inhibit either the expression or function of this potential tumour suppressor in lung cancers that retain the RBM5 gene.

  1. Contribution of Drosophila TRPA1-expressing neurons to circadian locomotor activity patterns.

    Directory of Open Access Journals (Sweden)

    Youngseok Lee

    Full Text Available In both vertebrates and invertebrates, Transient Receptor Potential (TRP channels are expressed in sensory neurons and mediate environmental stimuli such as light, sound, temperature, and taste. Some of these channels, however, are expressed only in the brain and their functions remain incompletely understood. Using the GAL4/UAS binary system with a line in which the GAL4 had been knocked into the trpA1 locus in Drosophila, we recently reported new insights into TRPA1 localization and function, including its expression in approximately 15% of all circadian neurons. TRPA1 is expressed in lateral posterior neurons (LPNs, which are known to be highly sensitive to entrainment by temperature cycles. Here, I used the bacterial sodium channel, NaChBac, to examine the effects of altering the electrical properties of trpA1 neurons on circadian rhythms. My results indicate that circadian activity of the flies in the morning, daytime, and evening was affected in a temperature-dependent manner following TRPA1 neuronal activation. Remarkably, TRPA1 neuron activation in flies kept at 18°C impacted the morning peak of circadian activity even though TRPA1 is not expressed in morning cells. Taken together, these results suggest that the activation of TRPA1-expressing neurons may differentially coordinate light/dark circadian entrainment, depending on the temperature.

  2. Contribution of Drosophila TRPA1-expressing neurons to circadian locomotor activity patterns.

    Science.gov (United States)

    Lee, Youngseok

    2013-01-01

    In both vertebrates and invertebrates, Transient Receptor Potential (TRP) channels are expressed in sensory neurons and mediate environmental stimuli such as light, sound, temperature, and taste. Some of these channels, however, are expressed only in the brain and their functions remain incompletely understood. Using the GAL4/UAS binary system with a line in which the GAL4 had been knocked into the trpA1 locus in Drosophila, we recently reported new insights into TRPA1 localization and function, including its expression in approximately 15% of all circadian neurons. TRPA1 is expressed in lateral posterior neurons (LPNs), which are known to be highly sensitive to entrainment by temperature cycles. Here, I used the bacterial sodium channel, NaChBac, to examine the effects of altering the electrical properties of trpA1 neurons on circadian rhythms. My results indicate that circadian activity of the flies in the morning, daytime, and evening was affected in a temperature-dependent manner following TRPA1 neuronal activation. Remarkably, TRPA1 neuron activation in flies kept at 18°C impacted the morning peak of circadian activity even though TRPA1 is not expressed in morning cells. Taken together, these results suggest that the activation of TRPA1-expressing neurons may differentially coordinate light/dark circadian entrainment, depending on the temperature.

  3. Ultraviolet A Enhances Cathepsin L Expression and Activity via JNK Pathway in Human Dermal Fibroblasts

    Science.gov (United States)

    Xu, Qing-Fang; Zheng, Yue; Chen, Jian; Xu, Xin-Ya; Gong, Zi-Jian; Huang, Yun-Fen; Lu, Chun; Maibach, Howard I; Lai, Wei

    2016-01-01

    Background: Cathepsin L (CatL) is a cysteine protease with strong matrix degradation activity that contributes to photoaging. Mannose phosphate-independent sorting pathways mediate ultraviolet A (UVA)-induced alternate trafficking of CatL. Little is known about signaling pathways involved in the regulation of UVA-induced CatL expression and activity. This study aims to investigate whether a single UVA irradiation affects CatL expression and activity and whether mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) pathway is involved in the regulation of UVA-induced CatL expression and activity in human dermal fibroblasts (HDFs). Methods: Primary HDFs were exposed to UVA. Cell proliferation was determined by a cell counting kit. UVA-induced CatL production and activity were studied with quantitative real-time reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and fluorimetric assay in cell lysates collected on three consecutive days after irradiation. Time courses of UVA-activated JNK and p38MAPK signaling were examined by Western blotting. Effects of MAPK inhibitors and knockdown of Jun and Fos on UVA-induced CatL expression and activity were investigated by RT-PCR, Western blotting, and fluorimetric assay. Data were analyzed by one-way analysis of variance. Results: UVA significantly increased CatL gene expression, protein abundance, and enzymatic activity for three consecutive days after irradiation (F = 83.11, 56.14, and 71.19, respectively; all P < 0.05). Further investigation demonstrated phosphorylation of JNK and p38MAPK activated by UVA. Importantly, inactivation of JNK pathway significantly decreased UVA-induced CatL expression and activity, which were not affected by p38MAPK inhibition. Moreover, knockdown of Jun and Fos significantly attenuated basal and UVA-induced CatL expression and activity. Conclusions: UVA enhances CatL production and activity in HDFs, probably by activating JNK and downstreaming AP-1. These

  4. Ultraviolet A Enhances Cathepsin L Expression and Activity via JNK Pathway in Human Dermal Fibroblasts

    Institute of Scientific and Technical Information of China (English)

    Qing-Fang Xu; Yue Zheng; Jian Chen; Xin-Ya Xu; Zi-Jian Gong; Yun-Fen Huang; Chun Lu

    2016-01-01

    Background:Cathepsin L (CatL) is a cysteine protease with strong matrix degradation activity that contributes to photoaging.Mannose phosphate-independent sorting pathways mediate ultraviolet A (UVA)-induced alternate trafficking of CatL.Little is known about signaling pathways involved in the regulation of UVA-induced CatL expression and activity.This study aims to investigate whether a single UVA irradiation affects CatL expression and activity and whether mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) pathway is involved in the regulation of UVA-induced CatL expression and activity in human dermal fibroblasts (HDFs).Methods:Primary HDFs were exposed to UVA.Cell proliferation was determined by a cell counting kit.UVA-induced CatL production and activity were studied with quantitative real-time reverse transcription polymerase chain reaction (RT-PCR),Western blotting,and fluorimetric assay in cell lysates collected on three consecutive days after irradiation.Time courses of UVA-activated JNK and p38MAPK signaling were examined by Western blotting.Effects ofMAPK inhibitors and knockdown of Jun and Fos on UVA-induced CatL expression and activity were investigated by RT-PCR,Western blotting,and fluorimetric assay.Data were analyzed by one-way analysis of variance.Results:UVA significantly increased CatL gene expression,protein abundance,and enzymatic activity for three consecutive days after irradiation (F =83.11,56.14,and 71.19,respectively;all P < 0.05).Further investigation demonstrated phosphorylation of JNK and p38MAPK activated by UVA.Importantly,inactivation of JNK pathway significantly decreased UVA-induced CatL expression and activity,which were not affected by p38MAPK inhibition.Moreover,knockdown of Jun and Fos significantly attenuated basal and UVA-induced CatL expression and activity.Conclusions:UVA enhances CatL production and activity in HDFs,probably by activating JNK and downstreaming AP-1.These findings provide a new possible

  5. Expression of the Inhibitory CD200 Receptor Is Associated with Alternative Macrophage Activation

    NARCIS (Netherlands)

    N. Koning; M. van Eijk; W. Pouwels; M.S.M. Brouwer; D. Voehringer; I. Huitinga; R.M. Hoek; G. Raes; J. Hamann

    2010-01-01

    Classical macrophage activation is inhibited by the CD200 receptor (CD200R). Here, we show that CD200R expression was specifically induced on human in vitro polarized macrophages of the alternatively activated M2a subtype, generated by incubation with IL-4 or IL-13. In mice, peritoneal M2 macrophage

  6. Temporal gene expression profile of human precursor B leukemia cells induced by adhesion receptor: identification of pathways regulating B-cell survival.

    Science.gov (United States)

    Astier, Anne Laurence; Xu, Ronghui; Svoboda, Marek; Hinds, Esther; Munoz, Olivier; de Beaumont, Rosalie; Crean, Colin Daniel; Gabig, Theodore; Freedman, Arnold Stephen

    2003-02-01

    The physical interactions between B cells and stromal cells from the lymphoid tissue microenvironment are critical to the survival of normal and malignant B cells. They are principally mediated by integrins expressed on B cells and counterreceptors on stromal cells. Specifically, alpha4beta1 integrin engagement rescues B cells from physiological or drug-induced apoptosis. Therefore, in order to understand the mechanisms by which integrins prevent apoptosis in leukemia B cells, we compared the temporal gene expression profiles induced by beta1-integrin ligation with fibronectin (Fn) or adhesion by poly-L-Lysine in serum-starved precursor B leukemia cells. Among the 38 selected differentially expressed genes, 6 genes involved in adhesion (VAV2, EPB41L1, CORO1A), proliferation (FRAP1, CCT4), and intercellular communication (GJB3) were validated by real-time quantitative polymerase chain reaction (RT-Q-PCR). Gene expression modulation could also be validated at the protein level for 5 other genes. We show that integrin stimulation up-regulated FBI-1 expression but inhibited CD79a, Requiem, c-Fos, and caspase 7 induction when the cells underwent apoptosis. We further demonstrate that Fn stimulation also inhibits caspase 3 activation but increases XIAP and survivin expression. Moreover, integrin stimulation also prevents caspase activation induced by doxorubicin. Therefore, we identified genes modulated by adhesion of human precursor B leukemia cells that regulate proliferation and apoptosis, highlighting new pathways that might provide insights into future therapy aiming at targeting apoptosis of leukemia cells.

  7. Effects of Diosgenin on Myometrial Matrix Metalloproteinase-2 and -9 Activity and Expression in Ovariectomized Rats

    Directory of Open Access Journals (Sweden)

    Chi-Chen Chang, Tang-Ching Kuan, Yao-Yuan Hsieh, Ying-Jui Ho, Yu-Ling Sun, Chih-Sheng Lin

    2011-01-01

    Full Text Available Diosgenin, a traditional Yam extraction, has been used in hormone replacement for menopausal women. We aimed to investigate the influences of diosgenin administration upon the MMP-2 and -9 activity and expression and reproductive hormones of ovariectomized (OVX rats, a model of menopausal status. Seven-week old female Wistar rats with bilateral OVX or sham operation (controls were divided and administered different dosages of diosgenin (0, 10, 50, or 100 mg/kg/day for 8 weeks. Serum was then sampled for progesterone (P4 and estradiol (E2 assay and uterine horns harvested. Myometrial MMP-2 and -9 activity and expression were surveyed and myometrial collagen expression was also assayed. The results show higher body weight in OVX rats across the 8 weeks post surgery and no significant differences were noted among OVX or Sham rats with diosgenin supplements. There were lower P4 and E2 concentrations in OVX rats compared to Sham rats, and higher P4 concentration of Sham rats post diosgenin supplement. MMP-2 and -9 mRNA expression and activity was lower in OVX rats, although higher MMP-2 and lower MMP-9 activity/mRNA expression was observed in OVX rats post diosgenin supplementation. Collagen mRNA expression was higher in OVX rats compared to Sham controls, and diosgenin administration decreased collagen mRNA expression in OVX rats. In conclusion, diosgenin is associated with gelatinase expression and collagen metabolism in OVX rats. Diosgenin administration can partially reverse the effects of OVX upon MMP functions and hormone status. Adequate diosgenin supplement might modulate myometrial gelatinase expression and collagen metabolism in menopausal subjects.

  8. Expression of a Deschampsia antarctica Desv. Polypeptide with Lipase Activity in a Pichia pastoris Vector

    Directory of Open Access Journals (Sweden)

    Claudia Rabert

    2014-02-01

    Full Text Available The current study isolated and characterized the Lip3F9 polypeptide sequence of Deschampsia antarctica Desv. (GeneBank Accession Number JX846628, which was found to be comprised of 291 base pairs and was, moreover, expressed in Pichia pastoris X-33 cells. The enzyme was secreted after 24 h of P. pastoris culture incubation and through induction with methanol. The expressed protein showed maximum lipase activity (35 U/L with an optimal temperature of 37 °C. The lipase-expressed enzyme lost 50% of its specific activity at 25 °C, a behavior characteristic of a psychrotolerant enzyme. Recombinant enzyme activity was measured in the presence of ionic and non-ionic detergents, and a decrease in enzyme activity was detected for all concentrations of ionic and non-ionic detergents assessed.

  9. Histidine tag fusion increases expression levels of active recombinant amelogenin in Escherichia coli.

    Science.gov (United States)

    Svensson, Johan; Andersson, Christer; Reseland, Janne E; Lyngstadaas, Petter; Bülow, Leif

    2006-07-01

    Amelogenin is a dental enamel matrix protein involved in formation of dental enamel. In this study, we have expressed two different recombinant murine amelogenins in Escherichia coli: the untagged rM179, and the histidine tagged rp(H)M180, identical to rM179 except that it carries the additional N-terminal sequence MRGSHHHHHHGS. The effects of the histidine tag on expression levels, and on growth properties of the amelogenin expressing cells were studied. Purification of a crude protein extract containing rp(H)M180 was also carried out using IMAC and reverse-phase HPLC. The results of this study showed clearly that both growth properties and amelogenin expression levels were improved for E. coli cells expressing the histidine tagged amelogenin rp(H)M180, compared to cells expressing the untagged amelogenin rM179. The positive effect of the histidine tag on amelogenin expression is proposed to be due to the hydrophilic nature of the histidine tag, generating a more hydrophilic amelogenin, which is more compatible with the host cell. Human osteoblasts treated with the purified rp(H)M180 showed increased levels of secreted osteocalcin, compared to untreated cells. This response was similar to cells treated with enamel matrix derivate, mainly composed by amelogenin, suggesting that the recombinant protein is biologically active. Thus, the histidine tag favors expression and purification of biologically active recombinant amelogenin.

  10. Matrine inhibits the expression of adhesion molecules in activated vascular smooth muscle cells.

    Science.gov (United States)

    Liu, Jun; Zhang, Lihua; Ren, Yingang; Gao, Yanli; Kang, Li; Lu, Shaoping

    2016-03-01

    Atherosclerosis is a chronic inflammatory disease associated with increased expression of adhesion molecules in vascular smooth muscle cells (VSMCs). Matrine is a main active ingredient of Sophora flavescens roots, which are used to treat inflammatory diseases. However, the effects of matrine on the expression of adhesion molecules in VSMCs have largely remained elusive. Therefore, the present study investigated the effects of matrine on the expression of adhesion molecules in tumor necrosis factor (TNF)‑α‑stimulated human aortic smooth muscle cells (HASMCs). The results showed that matrine inhibited the expression of vascular cell adhesion molecule‑1 (VCAM‑1) and intercellular adhesion molecule‑1 (ICAM‑1) in TNF‑α‑stimulated HASMCs. Matrine markedly inhibited the TNF‑α‑induced expression of nuclear factor (NF)‑κB p65 and prevented the TNF‑α‑caused degradation of inhibitor of NF‑κB; it also inhibited TNF‑α‑induced activation of mitogen‑activated protein kinases (MAPKs). Furthermore, matrine inhibited the production of intracellular reactive oxygen species (ROS) in TNF‑α‑stimulated HASMCs. In conclusion, the results of the present study demonstrated that matrine inhibited the expression of VCAM‑1 and ICAM‑1 in TNF‑α‑stimulated HASMCs via the suppression of ROS production as well as NF‑κB and MAPK pathway activation. Therefore, matrine may have a potential therapeutic use for preventing the advancement of atherosclerotic lesions.

  11. 5'-AMP-activated protein kinase activity and subunit expression in exercise-trained human skeletal muscle

    DEFF Research Database (Denmark)

    Nielsen, Jakob Nis; Mustard, Kirsty J.W.; Graham, Drew A.

    2002-01-01

    5'-AMP-activated protein kinase (AMPK) has been proposed to be a pivotal factor in cellular responses to both acute exercise and exercise training. To investigate whether protein levels and gene expression of catalytic (alpha(1), alpha(2)) and regulatory (beta(1), beta(2), gamma(1), gamma(2), gam...... muscle has increased alpha(1)-AMPK protein levels and blunted AMPK activation during exercise.......5'-AMP-activated protein kinase (AMPK) has been proposed to be a pivotal factor in cellular responses to both acute exercise and exercise training. To investigate whether protein levels and gene expression of catalytic (alpha(1), alpha(2)) and regulatory (beta(1), beta(2), gamma(1), gamma(2), gamma......(3)) AMPK subunits and exercise-induced AMPK activity are influenced by exercise training status, muscle biopsies were obtained from seven endurance exercise-trained and seven sedentary young healthy men. The alpha(1)- and alpha(2)-AMPK mRNA contents in trained subjects were both 117 +/- 2...

  12. DNA end binding activity and Ku70/80 heterodimer expression in human colorectal tumor

    Institute of Scientific and Technical Information of China (English)

    Paola Mazzarelli; Carolina Gravina; Marco Caricato; Maria Luana Poeta; Monica Rinaldi; Sergio Valeri; Roberto Coppola; Vito Michele Fazio; Paola Parrella; Davide Seripa; Emanuela Signori; Giuseppe Perrone; Carla Rabitti; Domenico Borzomati; Armando Gabbrielli; Maria Giovanna Matera

    2005-01-01

    AIM: To determine the DNA binding activity and protein levels of the Ku70/80 heterodimer, the functional mediator of the NHEJ activity, in human colorectal carcinogenesis.METHODS: The Ku70/80 DNA-binding activity was determined by electrophoretic mobility shift assays in 20 colon adenoma and 15 colorectal cancer samples as well as matched normal colonic tissues. Nuclear and cytoplasmic protein expression was determined by immunohistochemistry and Western blot analysis.RESULTS: A statistically significant difference was found in both adenomas and carcinomas as compared to matched normal colonic mucosa (P<0.00). However,changes in binding activity were not homogenous with approximately 50% of the tumors showing a clear increase in the binding activity, 30% displaying a modest increase and 15% showing a decrease of the activity.Tumors, with increased DNA-binding activity, also showed a statistically significant increase in Ku70 and Ku86nuclear expression, as determined by Western blot and immunohistochemical analyses (P<0.001). Cytoplasmic protein expression was found in pathological samples,but not in normal tissues either from tumor patients or from healthy subjects.CONCLUSION: Overall, our DNA-binding activity and protein level are consistent with a substantial activation of the NHEJ pathway in colorectal tumors. Since the NHEJ is an error prone mechanism, its abnormal activation can result in chromosomal instability and ultimately lead to tumorigenesis.

  13. TELOMERASE ACTIVITY AND hTERT mRNA EXPRESSION IN ACUTE LEUKEMIA

    Institute of Scientific and Technical Information of China (English)

    何冬梅; 张洹

    2004-01-01

    Objective: To investigate the clinical implications of telomerase activity and human telomerase reverse transcriptase (hTERT) expression as useful diagnostic marker in acute leukemia. Methods: Expression of hTERT was detected by reverse transcription- polymerase chain reaction (RT-PCR) in 24 cases with acute leukemia and in 12 normal persons. Quantitative levels of telomerase activity were examined by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA). Results: In the bone marrow and peripheral blood of 24 acute leukemia, telomerase activity was detected in 75% of the samples, with absorbances (A) of 0.538(0.062 and 0.463(0.054, respectively. Whereas in 12 normal peripheral blood, telomerase activity had only a positive rate of 8.3%, with A value of 0.16(0.012. telomerase activities in the bone marrow and peripheral blood of acute leukemia were significantly higher than in normal control (P<0.05). RT-PCR analysis revealed that hTERT mRNA was expressed in 79.17%(19/24) of acute leukemia, but in only 1 of 12 normal peripheral blood. In 24 acute leukemias, 17 cases had both positive telomerse activity and hTERT mRNA expression. The expression of hTERT mRNA is correlated with telomerase activity (P<0.01). Conclusion: Telomerase and hTERT mRNA could be useful in diagnosis of acute leukemia. hTERT gene expression was strongly associated with telomerase activity in acute leukemia.

  14. Tumour microenvironments induce expression of urokinase plasminogen activator receptor (uPAR and concomitant activation of gelatinolytic enzymes.

    Directory of Open Access Journals (Sweden)

    Synnøve Magnussen

    Full Text Available The urokinase plasminogen activator receptor (uPAR is associated with poor prognosis in oral squamous cell carcinoma (OSCC, and increased expression of uPAR is often found at the invasive tumour front. The aim of the current study was to elucidate the role of uPAR in invasion and metastasis of OSCC, and the effects of various tumour microenvironments in these processes. Furthermore, we wanted to study whether the cells' expression level of uPAR affected the activity of gelatinolytic enzymes.The Plaur gene was both overexpressed and knocked-down in the murine OSCC cell line AT84. Tongue and skin tumours were established in syngeneic mice, and cells were also studied in an ex vivo leiomyoma invasion model. Soluble factors derived from leiomyoma tissue, as well as purified extracellular matrix (ECM proteins, were assessed for their ability to affect uPAR expression, glycosylation and cleavage. Activity of gelatinolytic enzymes in the tissues were assessed by in situ zymography.We found that increased levels of uPAR did not induce tumour invasion or metastasis. However, cells expressing low endogenous levels of uPAR in vitro up-regulated uPAR expression both in tongue, skin and leiomyoma tissue. Various ECM proteins had no effect on uPAR expression, while soluble factors originating from the leiomyoma tissue increased both the expression and glycosylation of uPAR, and possibly also affected the proteolytic processing of uPAR. Tumours with high levels of uPAR, as well as cells invading leiomyoma tissue with up-regulated uPAR expression, all displayed enhanced activity of gelatinolytic enzymes.Although high levels of uPAR are not sufficient to induce invasion and metastasis, the activity of gelatinolytic enzymes was increased. Furthermore, several tumour microenvironments have the capacity to induce up-regulation of uPAR expression, and soluble factors in the tumour microenvironment may have an important role in the regulation of posttranslational

  15. SWI/SNF enzymes promote SOX10- mediated activation of myelin gene expression.

    Science.gov (United States)

    Marathe, Himangi G; Mehta, Gaurav; Zhang, Xiaolu; Datar, Ila; Mehrotra, Aanchal; Yeung, Kam C; de la Serna, Ivana L

    2013-01-01

    SOX10 is a Sry-related high mobility (HMG)-box transcriptional regulator that promotes differentiation of neural crest precursors into Schwann cells, oligodendrocytes, and melanocytes. Myelin, formed by Schwann cells in the peripheral nervous system, is essential for propagation of nerve impulses. SWI/SNF complexes are ATP dependent chromatin remodeling enzymes that are critical for cellular differentiation. It was recently demonstrated that the BRG1 subunit of SWI/SNF complexes activates SOX10 expression and also interacts with SOX10 to activate expression of OCT6 and KROX20, two transcriptional regulators of Schwann cell differentiation. To determine the requirement for SWI/SNF enzymes in the regulation of genes that encode components of myelin, which are downstream of these transcriptional regulators, we introduced SOX10 into fibroblasts that inducibly express dominant negative versions of the SWI/SNF ATPases, BRM or BRG1. Dominant negative BRM and BRG1 have mutations in the ATP binding site and inhibit gene activation events that require SWI/SNF function. Ectopic expression of SOX10 in cells derived from NIH 3T3 fibroblasts led to the activation of the endogenous Schwann cell specific gene, myelin protein zero (MPZ) and the gene that encodes myelin basic protein (MBP). Thus, SOX10 reprogrammed these cells into myelin gene expressing cells. Ectopic expression of KROX20 was not sufficient for activation of these myelin genes. However, KROX20 together with SOX10 synergistically activated MPZ and MBP expression. Dominant negative BRM and BRG1 abrogated SOX10 mediated activation of MPZ and MBP and synergistic activation of these genes by SOX10 and KROX20. SOX10 was required to recruit BRG1 to the MPZ locus. Similarly, in immortalized Schwann cells, BRG1 recruitment to SOX10 binding sites at the MPZ locus was dependent on SOX10 and expression of dominant negative BRG1 inhibited expression of MPZ and MBP in these cells. Thus, SWI/SNF enzymes cooperate with SOX10 to

  16. SWI/SNF enzymes promote SOX10- mediated activation of myelin gene expression.

    Directory of Open Access Journals (Sweden)

    Himangi G Marathe

    Full Text Available SOX10 is a Sry-related high mobility (HMG-box transcriptional regulator that promotes differentiation of neural crest precursors into Schwann cells, oligodendrocytes, and melanocytes. Myelin, formed by Schwann cells in the peripheral nervous system, is essential for propagation of nerve impulses. SWI/SNF complexes are ATP dependent chromatin remodeling enzymes that are critical for cellular differentiation. It was recently demonstrated that the BRG1 subunit of SWI/SNF complexes activates SOX10 expression and also interacts with SOX10 to activate expression of OCT6 and KROX20, two transcriptional regulators of Schwann cell differentiation. To determine the requirement for SWI/SNF enzymes in the regulation of genes that encode components of myelin, which are downstream of these transcriptional regulators, we introduced SOX10 into fibroblasts that inducibly express dominant negative versions of the SWI/SNF ATPases, BRM or BRG1. Dominant negative BRM and BRG1 have mutations in the ATP binding site and inhibit gene activation events that require SWI/SNF function. Ectopic expression of SOX10 in cells derived from NIH 3T3 fibroblasts led to the activation of the endogenous Schwann cell specific gene, myelin protein zero (MPZ and the gene that encodes myelin basic protein (MBP. Thus, SOX10 reprogrammed these cells into myelin gene expressing cells. Ectopic expression of KROX20 was not sufficient for activation of these myelin genes. However, KROX20 together with SOX10 synergistically activated MPZ and MBP expression. Dominant negative BRM and BRG1 abrogated SOX10 mediated activation of MPZ and MBP and synergistic activation of these genes by SOX10 and KROX20. SOX10 was required to recruit BRG1 to the MPZ locus. Similarly, in immortalized Schwann cells, BRG1 recruitment to SOX10 binding sites at the MPZ locus was dependent on SOX10 and expression of dominant negative BRG1 inhibited expression of MPZ and MBP in these cells. Thus, SWI/SNF enzymes cooperate

  17. Platelet activating factor-induced expression of p21 is correlated with histone acetylation

    Science.gov (United States)

    Damiani, Elisabetta; Puebla-Osorio, Nahum; Lege, Bree M.; Liu, Jingwei; Neelapu, Sattva S.; Ullrich, Stephen E.

    2017-01-01

    Ultraviolet (UV)-irradiated keratinocytes secrete the lipid mediator of inflammation, platelet-activating factor (PAF). PAF plays an essential role in UV-induced immune suppression and skin cancer induction. Dermal mast cell migration from the skin to the draining lymph nodes plays a prominent role in activating systemic immune suppression. UV-induced PAF activates mast cell migration by up-regulating mast cell CXCR4 surface expression. Recent findings indicate that PAF up-regulates CXCR4 expression via histone acetylation. UV-induced PAF also activates cell cycle arrest and disrupts DNA repair, in part by increasing p21 expression. Do epigenetic alterations play a role in p21 up-regulation? Here we show that PAF increases Acetyl-CREB-binding protein (CBP/p300) histone acetyltransferase expression in a time and dose-dependent fashion. Partial deletion of the HAT domain in the CBP gene, blocked these effects. Chromatin immunoprecipitation assays indicated that PAF-treatment activated the acetylation of the p21 promoter. PAF-treatment had no effect on other acetylating enzymes (GCN5L2, PCAF) indicating it is not a global activator of histone acetylation. This study provides further evidence that PAF activates epigenetic mechanisms to affect important cellular processes, and we suggest this bioactive lipid can serve as a link between the environment and the epigenome. PMID:28157211

  18. Ontogenic timing mechanism initiates the expression of rat intestinal sucrase activity

    Energy Technology Data Exchange (ETDEWEB)

    Yeh, K.Y.; Holt, P.R.

    1986-03-01

    Morphologic and enzymic differentiation occurs in rat small intestinal epithelium during 16-20 days of postnatal life. This change is considered to be initiated by an ontogenic timing mechanism and is modulated by extrinsic systemic and luminal factors. The importance of the ontogenic timing was tested directly using a transplantation technique in which jejunal isografts from newborn (day 0) and 5-day-old (day 5) rats were implanted under the skin of newborn (day 0) hosts. Isografts showing cryptvillus architecture were obtained in 44% and 21% of transplants, respectively. Day 0 isografts and host intestine expressed sucrase activity at about 16-18 days of age and showed similar crypt cell labeling and epithelial migration after (3H)thymidine injection. Day 5 isografts expressed sucrase activity when the hosts were 13 days of age, whereas host intestine showed no detectable sucrase activity. Isograft lactase activities in both experimental transplant models were significantly higher than host intestinal lactase up to 28 days of age, suggesting that luminal factors are important in modulating lactase activity during the first 4 wk of postnatal life. It is concluded that (a) no systemic factors at day 13 inhibit the expression of sucrase activity and (b) an ontogenic timing mechanism in the jejunum initiates the expression of sucrase activity.

  19. Radioprotection of 1,2-dimethylhydrazine-initiated colon cancer in rats using low-dose γ rays by modulating multidrug resistance-1, cytokeratin 20, and β-catenin expression.

    Science.gov (United States)

    Nabil, H M; Hassan, B N; Tohamy, A A; Waaer, H F; Abdel Moneim, A E

    2016-03-01

    Ionizing radiation is a widely used therapy for solid tumors. However, high-dose ionizing radiation causes apoptosis, transforms normal cells into tumor cells, and impairs immune functions, leading to the defects in the removal of damaged or tumor cells. In contrast, low-dose radiation has been reported to exert various beneficial effects in cells. This experimental study investigated the effect of γ rays at low dose on the development of colorectal tumor in a 1,2-dimethylhydrazine (DMH)-induced colon cancer. Colorectal tumor model was induced in Wistar rats by subcutaneous injection of DMH (20 mg/kg) once a week for 15 weeks. Starting from zero day of DMH injection, a single low dose of whole-body γ irradiation of 0.5 Gy/week was applied to the rats. A significant reduction in lipid peroxidation, nitric oxide, and elevation in the glutathione content and antioxidant enzyme activity (superoxide dismutase and catalase) were observed after γ irradiation comparing with DMH group. Moreover, γ ray reduced the expressions of multidrug resistance 1 (MDR1), β-catenin, and cytokeratin 20 (CK20) those increased in DMH-treated rats. However, survivin did not change with γ ray treatment. A histopathological examination of the DMH-injected rats revealed ulcerative colitis, dysplasia, anaplasia, and hyperchromasia. An improvement in the histopathological picture was seen in the colon of rats exposed to γ rays. In conclusion, the present results showed that low-dose γ ray significantly inhibited DMH-induced colon carcinogenesis in rats by modulating CK20, MDR1, and β-catenin expression but not survivin expression.

  20. Identification of chemical modulators of the constitutive activated receptor (CAR) in a gene expression compendium

    OpenAIRE

    Oshida, Keiyu; Vasani, Naresh; Jones, Carlton; Moore, Tanya; Hester, Susan; Nesnow, Stephen; Auerbach, Scott; Geter, David R.; Aleksunes, Lauren M; Thomas, Russell S.; Applegate, Dawn; Klaassen, Curtis D.; Corton, J. Christopher

    2015-01-01

    The nuclear receptor family member constitutive activated receptor (CAR) is activated by structurally diverse drugs and environmentally-relevant chemicals leading to transcriptional regulation of genes involved in xenobiotic metabolism and transport. Chronic activation of CAR increases liver cancer incidence in rodents, whereas suppression of CAR can lead to steatosis and insulin insensitivity. Here, analytical methods were developed to screen for chemical treatments in a gene expression comp...

  1. Activation profiles of opioid ligands in HEK cells expressing δ opioid receptors

    OpenAIRE

    Clark J; Demirci Hasan; Gharagozlou Parham; Lameh Jelveh

    2002-01-01

    Abstract Background The aim of the present study was to characterize the activation profiles of 15 opioid ligands in transfected human embryonic kidney cells expressing only δ opioid receptors. Activation profiles of most of these ligands at δ opioid receptors had not been previously characterized in vitro. Receptor activation was assessed by measuring the inhibition of forskolin-stimulated cAMP production. Results Naltrexone and nalorphine were classified as antagonists at δ opioid receptor....

  2. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard

    , membrane-bound Hsp70 can stimulate antigen presenting cells (APCs) to release proinflammatory cytokines and can provide a target structure for NK cell-mediated lysis. Human cancer cells frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several...... clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 surface expression on cancer cells. We have found that inhibition of histone deacetylase (HDAC) activity leads to surface expression of Hsp70 on various...... hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC-inhibitor mediated Hsp70 surface expression was confined to the apoptotic Annexin V positive cells and blocked by inhibition of apoptosis. Other chemotherapeutic inducers of apoptosis...

  3. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle

    -derived antigenic peptides, a function which is currently explored in immunotherapeutic approaches against cancer. Additionally, membrane-bound Hsp70 can stimulate antigen presenting cells to release proinflammatory cytokines and can provide a target structure for NK cell-mediated lysis. Human cancer cells...... frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 cell surface expression on cancer...... cells. We have found that inhibition of histone deacetylase (HDAC) activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC-inhibitor mediated Hsp70 cell surface expression was confined...

  4. Transient expression of organophosphorus hydrolase to enhance the degrading activity of tomato fruit on coumaphos

    Institute of Scientific and Technical Information of China (English)

    Jie-hong ZHAO; De-gang ZHAO

    2009-01-01

    We constructed an expression cassette of the organophosphorus pesticide degrading (opd)gene under the control of the E8 promoter.Then opd was transformed into tomato fruit using an agroinfiltration transient expression system.β-Glueuronidase (GUS) staining,reverse transcription-polymerase chain reaction (RT-PCR),wavelength scanning,and fluorescent reaction were performed to examine the expression of the opd gene and the hydrolysis activity on eoumaphos of organophosphorus hydrolase (OPH) in tomato fruit.The results show that the agroinfiltrated tomato fruit-expressed OPH had the maximum hydrolysis activity of about 11.59 U/mg total soluble protein.These results will allow us to focus on breeding transgenie plants that could not only enhance the degrading capability of fruit and but also hold no negative effects on pest control when spraying organophosphorus pesticides onto the seedlings in fields.

  5. Expression levels of microRNAs are not associated with their regulatory activities

    Directory of Open Access Journals (Sweden)

    Wu Jiarui

    2011-09-01

    Full Text Available Abstract MicroRNAs (miRNAs regulate their targets by triggering mRNA degradation or translational repression. The negative relationship between miRNAs and their targets suggests that the regulatory effect of a miRNA could be determined from the expression levels of its targets. Here, we investigated the relationship between miRNA activities determined by computational programs and miRNA expression levels by using data in which both mRNA and miRNA expression from the same samples were measured. We found that different from the intuitive expectation one might have, miRNA activity shows very weak correlation with miRNA expression, which indicates complex regulating mechanisms between miRNAs and their target genes. Reviewers This manuscript was reviewed by an anonymous reviewer and Dr Yuriy Gusev.

  6. Effects of high hydrostatic pressure on genomic expression profiling of porcine parthenogenetic activated and cloned embryos

    DEFF Research Database (Denmark)

    Lin, Lin; Luo, Yonglun; Sørensen, Peter;

    2014-01-01

    mechanism underlying the effects of HHP treatment on embryonic development is poorly understood and so was investigated in the present study. Thus, in the present study, we undertook genome-wide gene expression analysis in HHP-treated and untreated oocytes, as well as in 4-cell and blastocyst stage embryos...... derived by PA or HMC. Hierarchical clustering depicted stage-specific genomic expression profiling. At the 4-cell and blastocyst stages, 103 and 163 transcripts were differentially expressed between the HMC and PA embryos, respectively (P ... (INHBB and ME3) were further validated by quantitative reverse transcription–polymerase chain reaction. We also observed that HHP treatment activated expression of the imprinting gene DLX5 in 4-cell PA embryos. In conclusion, our genomic expression profiling data suggest that HHP alters the RNA...

  7. Modulation of perception and brain activity by predictable trajectories of facial expressions.

    Science.gov (United States)

    Furl, N; van Rijsbergen, N J; Kiebel, S J; Friston, K J; Treves, A; Dolan, R J

    2010-03-01

    People track facial expression dynamics with ease to accurately perceive distinct emotions. Although the superior temporal sulcus (STS) appears to possess mechanisms for perceiving changeable facial attributes such as expressions, the nature of the underlying neural computations is not known. Motivated by novel theoretical accounts, we hypothesized that visual and motor areas represent expressions as anticipated motion trajectories. Using magnetoencephalography, we show predictable transitions between fearful and neutral expressions (compared with scrambled and static presentations) heighten activity in visual cortex as quickly as 165 ms poststimulus onset and later (237 ms) engage fusiform gyrus, STS and premotor areas. Consistent with proposed models of biological motion representation, we suggest that visual areas predictively represent coherent facial trajectories. We show that such representations bias emotion perception of subsequent static faces, suggesting that facial movements elicit predictions that bias perception. Our findings reveal critical processes evoked in the perception of dynamic stimuli such as facial expressions, which can endow perception with temporal continuity.

  8. High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoris with Antimicrobial Activity

    Science.gov (United States)

    Iglesias-Figueroa, Blanca; Valdiviezo-Godina, Norberto; Siqueiros-Cendón, Tania; Sinagawa-García, Sugey; Arévalo-Gallegos, Sigifredo; Rascón-Cruz, Quintín

    2016-01-01

    In this study, bovine lactoferrin (bLf), an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf) in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin) demonstrated antibacterial activity against Escherichia coli (E. coli) BL21DE3, Staphylococcus aureus (S. aureus) FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa) ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly. PMID:27294912

  9. High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoris with Antimicrobial Activity

    Directory of Open Access Journals (Sweden)

    Blanca Iglesias-Figueroa

    2016-06-01

    Full Text Available In this study, bovine lactoferrin (bLf, an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin demonstrated antibacterial activity against Escherichia coli (E. coli BL21DE3, Staphylococcus aureus (S. aureus FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly.

  10. Gene expression changes in rat prostate after activation or blocking of the androgen and estrogen receptor

    DEFF Research Database (Denmark)

    Nellemann, Christine Lydia; Dalgaard, Majken; Holst, Bjørn;

    2005-01-01

    Several endpoints of different molecular complexity were studied in the Hershberger assay in order to evaluate the specificity and suitability of this test as a broad screening model. Androgen and estrogen receptors were activated or blocked, and expression of typical estrogen- or androgen...... and the anti-estrogen, ICI 182780, only affected ODC expression. Therefore, estrogenic or anti-estrogenic compounds would not be expected to seriously affect the outcome of a Hershberger test. However, EB given alone to castrated rats resulted in various effects. EB increased seminal vesicle weight, an effect...... reversed by ICI 182780, and affected TRPM-2, PBP C3, ODC, IGF-1, AR, and ERa mRNA levels. AR expression in the prostate seemed to be under regulation of both estrogens and androgens, as ICI 182780 inhibited the testosterone-induced AR expression, and flutamide inhibited the EB-induced AR expression...

  11. Sex differences in neural activation to facial expressions denoting contempt and disgust.

    Directory of Open Access Journals (Sweden)

    André Aleman

    Full Text Available The facial expression of contempt has been regarded to communicate feelings of moral superiority. Contempt is an emotion that is closely related to disgust, but in contrast to disgust, contempt is inherently interpersonal and hierarchical. The aim of this study was twofold. First, to investigate the hypothesis of preferential amygdala responses to contempt expressions versus disgust. Second, to investigate whether, at a neural level, men would respond stronger to biological signals of interpersonal superiority (e.g., contempt than women. We performed an experiment using functional magnetic resonance imaging (fMRI, in which participants watched facial expressions of contempt and disgust in addition to neutral expressions. The faces were presented as distractors in an oddball task in which participants had to react to one target face. Facial expressions of contempt and disgust activated a network of brain regions, including prefrontal areas (superior, middle and medial prefrontal gyrus, anterior cingulate, insula, amygdala, parietal cortex, fusiform gyrus, occipital cortex, putamen and thalamus. Contemptuous faces did not elicit stronger amygdala activation than did disgusted expressions. To limit the number of statistical comparisons, we confined our analyses of sex differences to the frontal and temporal lobes. Men displayed stronger brain activation than women to facial expressions of contempt in the medial frontal gyrus, inferior frontal gyrus, and superior temporal gyrus. Conversely, women showed stronger neural responses than men to facial expressions of disgust. In addition, the effect of stimulus sex differed for men versus women. Specifically, women showed stronger responses to male contemptuous faces (as compared to female expressions, in the insula and middle frontal gyrus. Contempt has been conceptualized as signaling perceived moral violations of social hierarchy, whereas disgust would signal violations of physical purity. Thus, our

  12. Generalised expression for the determination of activation energy of a thermoluminescence peak

    Energy Technology Data Exchange (ETDEWEB)

    Singh, S.J.; Gartia, R.K.; Mazumdar, P.S.

    1989-03-14

    A new set of expressions has been derived for the determination of the activation energy E of a thermoluminescence (TL) peak. Unlike those previously existing in the literature, the present expressions take into account the variation of the symmetry factor ..mu..g(x) with u/sub m/=(E/kT/sub m/) where x is the fractional intensity, k the Boltzmann constant and T/sub m/ the peak temperature and prior knowledge of the order of kinetics b is not required. The applicability of the present expressions have been tested using both numerically generated and experimental thermoluminescence peaks.

  13. Adolescent Mouse Takes on An Active Transcriptomic Expression During Postnatal Cerebral Development

    Institute of Scientific and Technical Information of China (English)

    Wei Xu; Chengqi Xin; Qiang Lin; Feng Ding; Wei Gong; Yuanyuan Zhou; Jun Yu; Peng Cui; Songnian Hu

    2014-01-01

    Postnatal cerebral development is a complicated biological process precisely controlled by multiple genes. To understand the molecular mechanism of cerebral development, we compared dynamics of mouse cerebrum transcriptome through three developmental stages using high-throughput RNA-seq technique. Three libraries were generated from the mouse cerebrum at infancy, adolescence and adulthood, respectively. Consequently, 44,557,729 (infancy), 59,257,530 (adolescence) and 72,729,636 (adulthood) reads were produced, which were assembled into 15,344, 16,048 and 15,775 genes, respectively. We found that the overall gene expression level increased from infancy to adolescence and decreased later on upon reaching adulthood. The adolescence cerebrum has the most active gene expression, with expression of a large number of reg-ulatory genes up-regulated and some crucial pathways activated. Transcription factor (TF) analysis suggested the similar dynamics as expression profiling, especially those TFs functioning in neurogenesis differentiation, oligodendrocyte lineage determination and circadian rhythm regula-tion. Moreover, our data revealed a drastic increase in myelin basic protein (MBP)-coding gene expression in adolescence and adulthood, suggesting that the brain myelin may be generated since mouse adolescence. In addition, differential gene expression analysis indicated the activation of rhythmic pathway, suggesting the function of rhythmic movement since adolescence;Furthermore, during infancy and adolescence periods, gene expression related to axon repulsion and attraction showed the opposite trends, indicating that axon repulsion was activated after birth, while axon attraction might be activated at the embryonic stage and declined during the postnatal develop-ment. Our results from the present study may shed light on the molecular mechanism underlying the postnatal development of the mammalian cerebrum.

  14. Adolescent mouse takes on an active transcriptomic expression during postnatal cerebral development.

    Science.gov (United States)

    Xu, Wei; Xin, Chengqi; Lin, Qiang; Ding, Feng; Gong, Wei; Zhou, Yuanyuan; Yu, Jun; Cui, Peng; Hu, Songnian

    2014-06-01

    Postnatal cerebral development is a complicated biological process precisely controlled by multiple genes. To understand the molecular mechanism of cerebral development, we compared dynamics of mouse cerebrum transcriptome through three developmental stages using high-throughput RNA-seq technique. Three libraries were generated from the mouse cerebrum at infancy, adolescence and adulthood, respectively. Consequently, 44,557,729 (infancy), 59,257,530 (adolescence) and 72,729,636 (adulthood) reads were produced, which were assembled into 15,344, 16,048 and 15,775 genes, respectively. We found that the overall gene expression level increased from infancy to adolescence and decreased later on upon reaching adulthood. The adolescence cerebrum has the most active gene expression, with expression of a large number of regulatory genes up-regulated and some crucial pathways activated. Transcription factor (TF) analysis suggested the similar dynamics as expression profiling, especially those TFs functioning in neurogenesis differentiation, oligodendrocyte lineage determination and circadian rhythm regulation. Moreover, our data revealed a drastic increase in myelin basic protein (MBP)-coding gene expression in adolescence and adulthood, suggesting that the brain myelin may be generated since mouse adolescence. In addition, differential gene expression analysis indicated the activation of rhythmic pathway, suggesting the function of rhythmic movement since adolescence; Furthermore, during infancy and adolescence periods, gene expression related to axonrepulsion and attraction showed the opposite trends, indicating that axon repulsion was activated after birth, while axon attraction might be activated at the embryonic stage and declined during the postnatal development. Our results from the present study may shed light on the molecular mechanism underlying the postnatal development of the mammalian cerebrum.

  15. Adolescent Mouse Takes on An Active Transcriptomic Expression During Postnatal Cerebral Development

    KAUST Repository

    Xu, Wei

    2014-06-01

    Postnatal cerebral development is a complicated biological process precisely controlled by multiple genes. To understand the molecular mechanism of cerebral development, we compared dynamics of mouse cerebrum transcriptome through three developmental stages using high-throughput RNA-seq technique. Three libraries were generated from the mouse cerebrum at infancy, adolescence and adulthood, respectively. Consequently, 44,557,729 (infancy), 59,257,530 (adolescence) and 72,729,636 (adulthood) reads were produced, which were assembled into 15,344, 16,048 and 15,775 genes, respectively. We found that the overall gene expression level increased from infancy to adolescence and decreased later on upon reaching adulthood. The adolescence cerebrum has the most active gene expression, with expression of a large number of regulatory genes up-regulated and some crucial pathways activated. Transcription factor (TF) analysis suggested the similar dynamics as expression profiling, especially those TFs functioning in neurogenesis differentiation, oligodendrocyte lineage determination and circadian rhythm regulation. Moreover, our data revealed a drastic increase in myelin basic protein (MBP)-coding gene expression in adolescence and adulthood, suggesting that the brain myelin may be generated since mouse adolescence. In addition, differential gene expression analysis indicated the activation of rhythmic pathway, suggesting the function of rhythmic movement since adolescence; Furthermore, during infancy and adolescence periods, gene expression related to axon. repulsion and attraction showed the opposite trends, indicating that axon repulsion was activated after birth, while axon attraction might be activated at the embryonic stage and declined during the postnatal development. Our results from the present study may shed light on the molecular mechanism underlying the postnatal development of the mammalian cerebrum. © 2014 .

  16. Ethanol Activation of PKA Mediates Single-Minded 2 Expression in Neuronal Cells.

    Science.gov (United States)

    Wang, Xiaolan; Yang, Zhihua; Sun, Yinan; Zhou, Hanjing; Chu, Guangpin; Zhang, Jing; Meng, Xianfang

    2015-12-01

    Prenatal ethanol exposure can cause extensive apoptotic neurodegeneration throughout the developing central nervous system (CNS), which results in cognitive deficits and memory decline. However, the underlying mechanisms need further study. Single-minded 2 (Sim2), a transcriptional repressor, is reportedly involved in diseases that impair learning and memory, such as Down syndrome (DS) and Alzheimer's disease. It is still unknown whether Sim2 is involved in regulating ethanol-mediated neuronal injury that might ultimately lead to neuronal dysfunction and subsequent learning and memory deficits. To study the effects of ethanol on Sim2 expression and neuronal injury, we used animal models and cell culture experiments. Our results indicated that in SH-SY5Y cells, ethanol exposure increased Sim2 expression and levels of cleaved caspase 3, which is a marker for cells undergoing apoptosis. Silencing Sim2 expression attenuated caspase 3 activation and cellular apoptosis. We also found that protein kinase A (PKA) activation induced Sim2 expression, as did ethanol. Inhibiting the PKA signaling pathway with H-89 decreased Sim2 expression and cleavage of caspase 3 that was induced by ethanol in vivo and in vitro. We further found that PKA regulated Sim2 expression at the transcriptional level. These results demonstrate that ethanol leads to increased Sim2 expression via the PKA pathway, ultimately resulting in apoptotic cell death.

  17. Prokaryotic Expression and Biological Activity Analysis of Human Ar-resten Gene

    Institute of Scientific and Technical Information of China (English)

    SONG Zifang; ZHENG Qichang; LI Wei; XIONG Jun; SHANG Dan; SHU Xiaogang

    2005-01-01

    To express recombinant arresten in Escherichia coli (E. Coli) and investigate its biological activity, prokaryotic expression vector of human arresten gene was constructed by gene engineering. Human arresten gene was amplified from recombinant plasmid pGEMArr by polymerase chain reaction (PCR), and inserted into prokaryotic expression vector pRSET containing T7 promoter. Restriction analysis and DNA sequencing verified that the arresten gene was correctly cloned into the expression vector. The recombinant plasmid pRSETAt was subsequently transformed into E. Coli BL21 (DE3), and the target gene was expressed under induction of IPTG. SDS-PAGE analysis revealed that the recombinant protein with a molecular weight of 29 kD (1 kD=0. 992 1 ku) amounted to 29 % of the total bacterial proteins. After purification and renaturation, the recombinant protein could significantly suppress the proliferation of human umbilical vein endothelial cells (HUVECs). These results suggested that the expression of a biologically active form of human arresten in the pRSET expression system laid a foundation for further study on the mechanistic insight into arresten action on angiogenesis and the development of powerful anti-cancer drugs.

  18. Inhibition of Nischarin Expression Promotes Neurite Outgrowth through Regulation of PAK Activity.

    Directory of Open Access Journals (Sweden)

    Yuemin Ding

    Full Text Available Nischarin is a cytoplasmic protein expressed in various organs that plays an inhibitory role in cell migration and invasion and the carcinogenesis of breast cancer cells. We previously reported that Nischarin is highly expressed in neuronal cell lines and is differentially expressed in the brain tissue of adult rats. However, the physiological function of Nischarin in neural cells remains unknown. Here, we show that Nischarin is expressed in rat primary cortical neurons but not in astrocytes. Nischarin is localized around the nucleus and dendrites. Using shRNA to knockdown the expression of endogenous Nischarin significantly increases the percentage of neurite-bearing cells, remarkably increases neurite length, and accelerates neurite extension in neuronal cells. Silencing Nischarin expression also promotes dendrite elongation in rat cortical neurons where Nischarin interacts with p21-activated kinase 1/2 (PAK1/2 and negatively regulates phosphorylation of both PAK1 and PAK2. The stimulation of neurite growth observed in cells with decreased levels of Nischarin is partially abolished by IPA3-mediated inhibition of PAK1 activity. Our findings indicate that endogenous Nischarin inhibits neurite outgrowth by blocking PAK1 activation in neurons.

  19. A novel baculovirus-derived promoter with high activity in the baculovirus expression system

    Directory of Open Access Journals (Sweden)

    María Martínez-Solís

    2016-06-01

    Full Text Available The baculovirus expression vector system (BEVS has been widely used to produce a large number of recombinant proteins, and is becoming one of the most powerful, robust, and cost-effective systems for the production of eukaryotic proteins. Nevertheless, as in any other protein expression system, it is important to improve the production capabilities of this vector. The orf46 viral gene was identified among the most highly abundant sequences in the transcriptome of Spodoptera exigua larvae infected with its native baculovirus, the S. exigua multiple nucleopolyhedrovirus (SeMNPV. Different sequences upstream of the orf46 gene were cloned, and their promoter activities were tested by the expression of the GFP reporter gene using the Autographa californica nucleopolyhedrovirus (AcMNPV vector system in different insect cell lines (Sf21, Se301, and Hi5 and in larvae from S. exigua and Trichoplusia ni. The strongest promoter activity was defined by a 120 nt sequence upstream of the ATG start codon for the orf46 gene. On average, GFP expression under this new promoter was more than two fold higher than the expression obtained with the standard polyhedrin (polh promoter. Additionally, the orf46 promoter was also tested in combination with the polh promoter, revealing an additive effect over the polh promoter activity. In conclusion, this new characterized promoter represents an excellent alternative to the most commonly used baculovirus promoters for the efficient expression of recombinant proteins using the BEVS.

  20. Antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus

    Directory of Open Access Journals (Sweden)

    Grabova A. Yu.

    2016-02-01

    Full Text Available Aim. To research the antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus. Methods. Deferred antagonism method, PCR, qRT-PCR, MALDI-TOF mass spectrometry. Results. It was revealed that Bacillus sp. strains C6 and Lg37s out of five tested strains had the highest antifungal activity. Based on the molecular genetic methods, it was shown that the expression of genes of lipopeptide antibiotics, related to the fengycin family, occurred in all these strains. At the same time, the gene expression of cyclolipopeptide iturin was found in the Bacillus sp. strains C6 and Lg37s. It was determined that Bacillus sp. C6 strain had the highest level of expression of the fengycin operon`s genes, whereas the lowest level was observed in Bacillus sp. C10 strain. By means of MALDI-TOF mass spectrometry, the presence of fengycins in the cell-free cultural fluid of Bacillus sp. C6 strain was detected. Conclusion. The direct correlation between the level of antifungal activity and the fengycin synthetases expression has not been disclosed. A higher level of antagonism detected for two Bacillus strains is more likely associated with the expression and subsequent synthesis of fengycin and iturin.