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Sample records for activation survivin expression

  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. Activation of Fms-like tyrosine kinase 3 signaling enhances survivin expression in a mouse model of rheumatoid arthritis.

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    Sofia E M Andersson

    Full Text Available Survivin is known as an inhibitor of apoptosis and a positive regulator of cell division. We have recently identified survivin as a predictor of joint destruction in patients with rheumatoid arthritis (RA. Flt3 ligand (Flt3L is expressed in the inflamed joints and has adjuvant properties in arthritis. Studies on 90 RA patients (median age 60.5 years [range, 24-87], disease duration 10.5 years [range, 0-35] show a strong positive association between the levels of survivin and Flt3L in blood. Here, we present experimental evidence connecting survivin and Flt3L signaling. Treatment of BALB/c mice with Flt3L led to an increase of survivin in the bone marrow and in splenic dendritic cells. Flt3L changed the profile of survivin splice variants, increasing transcription of the short survivin40 in the bone marrow. Treatment with an Flt3 inhibitor reduced total survivin expression in bone marrow and in the dendritic cell population in spleen. Inhibition of survivin transcription in mice, by shRNA lentiviral constructs, reduced the gene expression of Flt3L. We conclude that expression of survivin is a downstream event of Flt3 signaling, which serves as an essential mechanism supporting survival of leukocytes during their differentiation, and maturation of dendritic cells, in RA.

  4. Expression of survivin and its significance in colorectal cancer

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    Wei-Chang Chen; Qiang Liu; Jian-Xin Fu; Su-Ya Kang

    2004-01-01

    AIM: To study the expression of survivin,a novel member of inhibitors of apoptosis protein (IAP) and its significance in colorectal carcinoma.METHODS: Survivin mRNA expression was evaluated by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) in 52 colorectal carcinoma samples and48 adjacent normal colorectal tissue samples. PCR product was sequenced to verify the desired result. Expressions of survivin protein, proliferating cell nuclear antigen labelling index (PI) and apoptotic index (AI) were detected immunohistochemically in 52 human colorectal carcinomas.RESULTS: The expression of survivin mRNA was detected in a significantly greater proportion of colorectal carcinoma samples than in adjacent normal colorectal tissues (67.3%vs 25%; P<0.01). There was no relationship between survivin mRNA expression in colorectal carcinomas and sex, tumor size, histological types, lymphnode metastasis,distant metastasis and Dukes' stage. The PCR product shared 99% of homology with human counterparts. Survivin expression was observed immunohistochemically in 27 of52 cases of colorectal carcinoma (51.9%). The AI was significently lower in survivin positive group than in survivin negative group (0.67±0.18% vs1.14±0.42%; P<0.001),while the PI was greater in survivin positive group than in survivin negative group (51±22% vs 27±18%, P<0.001).CONCLUSION: Survivin is a special tumor marker independent of histopathological characteristics. It may play an important role during human colorectal tumorigenesis by inhibiting apoptosis and accelerating proliferative activity of colorectal tumor cells.

  5. 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.

  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. Inhibition of survivin influences the biological activities of canine histiocytic sarcoma cell lines.

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    Hiroki Yamazaki

    Full Text Available Canine histiocytic sarcoma (CHS is an aggressive malignant neoplasm that originates from histiocytic lineage cells, including dendritic cells and macrophages, and is characterized by progressive local infiltration and a very high metastatic potential. Survivin is as an apoptotic inhibitory factor that has major functions in cell proliferation, including inhibition of apoptosis and regulation of cell division, and is expressed in most types of human and canine malignant neoplasms, including melanoma and osteosarcoma. To investigate whether survivin was expressed at high levels in CHS and whether its expression was correlated with the aggressive biological behavior of CHS, we assessed relation between survivin expression and CHS progression, as well as the effects of survivin inhibition on the biological activities of CHS cells. We comparatively analyzed the expression of 6 selected anti-apoptotic genes, including survivin, in specimens from 30 dogs with histiocytic sarcoma and performed annexin V staining to evaluate apoptosis, methylthiazole tetrazolium assays to assess cell viability and chemosensitivity, and latex bead assays to measure changes in phagocytic activities in 4 CHS cell lines and normal canine fibroblasts transfected with survivin siRNA. Survivin gene expression levels in 30 specimens were significantly higher than those of the other 6 genes. After transfection with survivin siRNA, apoptosis, cell growth inhibition, enhanced chemosensitivity, and weakened phagocytic activities were observed in all CHS cell lines. In contrast, normal canine fibroblasts were not significantly affected by survivin knockdown. These results suggested that survivin expression may mediate the aggressive biological activities of CHS and that survivin may be an effective therapeutic target for the treatment of CHS.

  10. Cancer cell sensitivity to bortezomib is associated with survivin expression and p53 status but not cancer cell types

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    Chanan-Khan Asher A

    2010-01-01

    Full Text Available Abstract Background Survivin is known playing a role in drug resistance. However, its role in bortezomib-mediated inhibition of growth and induction of apoptosis is unclear. There are conflicting reports for the effect of bortezomib on survivin expression, which lacks of a plausible explanation. Methods: In this study, we tested cancer cells with both p53 wild type and mutant/null background for the relationship of bortezomib resistance with survivin expression and p53 status using MTT assay, flow cytometry, DNA fragmentation, caspase activation, western blots and RNAi technology. Results We found that cancer cells with wild type p53 show a low level expression of survivin and are sensitive to treatment with bortezomib, while cancer cells with a mutant or null p53 show a high level expression of survivin and are resistant to bortezomib-mediated apoptosis induction. However, silencing of survivin expression utilizing survivin mRNA-specific siRNA/shRNA in p53 mutant or null cells sensitized cancer cells to bortezomib mediated apoptosis induction, suggesting a role for survivin in bortezomib resistance. We further noted that modulation of survivin expression by bortezomib is dependent on p53 status but independent of cancer cell types. In cancer cells with mutated p53 or p53 null, bortezomib appears to induce survivin expression, while in cancer cells with wild type p53, bortezomib downregulates or shows no significant effect on survivin expression, which is dependent on the drug concentration, cell line and exposure time. Conclusions Our findings, for the first time, unify the current inconsistent findings for bortezomib treatment and survivin expression, and linked the effect of bortezomib on survivin expression, apoptosis induction and bortezomib resistance in the relationship with p53 status, which is independent of cancer cell types. Further mechanistic studies along with this line may impact the optimal clinical application of bortezomib in

  11. Survivin and related proteins in canine mammary tumors: immunohistochemical expression.

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    Bongiovanni, L; Romanucci, M; Malatesta, D; D'Andrea, A; Ciccarelli, A; Della Salda, L

    2015-03-01

    Survivin is reexpressed in most human breast cancers, where its expression has been associated with tumor aggressiveness, poor prognosis, and poor response to therapy. Survivin expression was evaluated in 41 malignant canine mammary tumors (CMTs) by immunohistochemistry, in relation to histological grade and stage, and correlated with that of some related molecules (β-catenin, caspase 3, heat shock proteins) to understand their possible role in canine mammary tumorigenesis. An increase in nuclear survivin expression, compared with healthy mammary glands, was observed in CMTs, where nuclear immunolabeling was related to the presence of necrosis. No statistically significant relation was found between the expression of the investigated molecules and the histological grade or stage. The present study may suggest an important involvement of survivin in CMT tumorigenesis. Its overexpression in most of the cases evaluated might suggest that targeting survivin in CMTs may be a valid anticancer therapy. PMID:24686389

  12. Expression of survivin protein in human colorectal carcinogenesis

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    Lian-Jie Lin; Chang-Qing Zheng; Yu Jin; Ying Ma; Wei-Guo Jiang; Tie Ma

    2003-01-01

    AIM: To identify the role of survivin in colorectal carcinogenesis and the relationship between Survivin and histological differentiation grade of colorectal carcinoma.METHODS: Immunohistochemical staining of survivin by using the monoclonal antibody was performed by the standard streptavidin-peroxidase (SP) technique for the 188paraffin sections which included 30 normal colorectal mucosas, 41 adenomas with low grade dysplasia, 30adenomas with high grade dysplasia, and 87 colorectal carcinomas which were classified as high, middle and low differentiated subgroups which included 33, 28, 26 cases respectively.RESULTS: Expression of survivin was observed in the cytoplasm of adenoma with dysplasia and colorectal carcinoma cells. No immunoreactivity of survivin was seen in normal mucosas. The positive rate of survivin increased in the transition from normal mucosas to adenomas with low grade dysplasia to high grade dysplasia/carcinomas (0.0 %, 31.7 %, 56.7 % and 63.2% respectively). But the difference between high grade dyspiasia and carcinomas had no statistical significance. Positive rate was not related to histological differentiation grade of colorectal carcinoma.Moreover, there was no correlation between histological differentiation grade of colorectal carcinoma and immunoreactive intensity of survivin.CONCLUSION: The expression of survivin is the essential event in the early stage of colorectal carcinogenesis and plays an important role in the transition sequence and it is not related to histological differentiation grade of colorectal carcinoma. It thus may provide a new diagnostic and therapeutic target in colorectal cancer.

  13. The prognostic significance of survivin expression in gallbladder carcinoma.

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    Salman, Tarik; Argon, Asuman; Kebat, Tulu; Vardar, Enver; Erkan, Nazif; Alacacıoğlu, Ahmet

    2016-08-01

    Gallbladder cancers (GBC) are characterized by rapid progression, early metastasis, and poor prognosis; the molecular mechanisms of the various signaling pathways involved should be elucidated to develop effective therapies. Survivin, an apoptosis inhibitor protein expressed in the G2/M phase of the cell cycle, plays a role in cell division and affects both cell survival and proliferation. Survivin has been investigated in many types of cancer, and this study aims to examine the relationship of survivin expression in gallbladder cancer patients with clinicopathological features and prognosis. We evaluated demographic characteristics (age, gender), tumor characteristics (histopathological type, differentiation, perineural, and lymphovascular invasion; serosal invasion, surgical margin positivity and lymphocytic response), and Survivin expression immunohistochemically, and we analysed the relationship between these characteristics and prognosis in 47 gallbladder carcinoma cases from 2000 to 2011. Immunohistochemically, while survivin expression was observed in 36 cases, it was absent in 11 cases. Follow-up data were obtained from 32 patients. Two (8.7%) of 23 cases with a Survivin-positive tumor were alive at 74th and 35th months, whereas 5 (%55.6) of nine cases with Survivin-negative tumor were alive at 50th, 89th, 124th, 126th, 131th months. Survivin expression was correlated with short survival (p = 0.043), and the univariate analysis showed that reduced overall survival was associated with age (p = 0.043), male gender (p = 0.038), infiltrative pattern (p = 0.019), lymphovascular invasion (p = 0.004), perineural invasion (p = 0.009), serosal invasion (p = 0.027), ulcer (p = 0.033), and surgical margin positivity (p = 0.022). Despite the low number of patients in our study, the analysis results suggest that survivin positivity might actually be a significant prognostic factor. This finding could be a reference point for targeted treatment studies. However, further

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. Prognostic significance of survivin expression in renal cell cancer and its correlation with radioresistance.

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    Lei, Yu; Geng, Zhang; Guo-Jun, Wu; He, Wang; Jian-Lin, Yuan

    2010-11-01

    Survivin, an important inhibitor of apoptosis, has been found to play an important role in the initiation, progression, and chemoradioresistance of human malignancies. Previously, we have reported that upregulation of survivin in oral squamous cell carcinoma correlates with poor prognosis and chemoresistance. The aim of this study was to assess prognostic significance of survivin protein expression in RCC and analyze its correlation with radiosensitivity of RCC cells. RT-PCR and Western blot assays were performed to detect survivin mRNA and protein expression in normal human kidney epithelial cell line (HKEC) or RCC cell lines. The expression of survivin mRNA in RCC and corresponding nontumor kidney tissues was also detected by RT-PCR. Immunohistochemistry was performed to determine survivin protein expression in 75 cases of RCC tissue samples. Moreover, the association of survivin protein expression with clinicopathogical factors and prognosis of RCC patients was statistically analyzed. Small interfering RNA was used to knockdown the endogenous survivin expression in RCC cell line (ACHN) and evaluate the effects of survivin knockdown on proliferation, apoptosis, and radiosensitivity of RCC cell line. RCC cells showed sufficient expression of survivin mRNA and protein, but the expression of survivin gene was not detected in normal HKEC. Moreover, the expression level of survivin mRNA in RCC tissues was significantly higher than that in corresponding nontumor kidney tissues. The immunostaining of survivin protein was mainly located in cytoplasm of RCC tumor cells. Tumor pathological stage (P = 0.028), grade (P = 0.004), and lymph node metastasis (P = 0.017) of RCC patients were significantly correlated with survivin protein expression. In addition, patients with high survivin levels had a significantly shorter overall survival than those with low levels (P < 0.001), and the expression of survivin protein was an independent prognostic factor for RCC patients (P = 0

  1. Epithelial PIK3R1 (p85) and TP53 Regulate Survivin Expression during Adaptation to Ileocecal Resection.

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    Cohran, Valeria; Managlia, Elizabeth; Bradford, Emily M; Goretsky, Tatiana; Li, Ting; Katzman, Rebecca B; Cheresh, Paul; Brown, Jeffrey B; Hawkins, Jennifer; Liu, Shirley X L; De Plaen, Isabelle G; Weitkamp, Jörn-Hendrik; Helmrath, Michael; Zhang, Zheng; Barrett, Terrence A

    2016-07-01

    Intestinal adaptation to small-bowel resection (SBR) after necrotizing enterocolitis expands absorptive surface areas and promotes enteral autonomy. Survivin increases proliferation and blunts apoptosis. The current study examines survivin in intestinal epithelial cells after ileocecal resection. Wild-type and epithelial Pik3r1 (p85α)-deficient mice underwent sham surgery or 30% resection. RNA and protein were isolated from small bowel to determine levels of β-catenin target gene expression, activated caspase-3, survivin, p85α, and Trp53. Healthy and post-resection human infant small-bowel sections were analyzed for survivin, Ki-67, and TP53 by immunohistochemistry. Five days after ileocecal resection, epithelial levels of survivin increased relative to sham-operated on mice, which correlated with reduced cleaved caspase-3, p85α, and Trp53. At baseline, p85α-deficient intestinal epithelial cells had less Trp53 and more survivin, and relative responses to resection were blunted compared with wild-type. In infant small bowel, survivin in transit amplifying cells increased 71% after SBR. Resection increased proliferation and decreased numbers of TP53-positive epithelial cells. Data suggest that ileocecal resection reduces p85α, which lowers TP53 activation and releases survivin promoter repression. The subsequent increase in survivin among transit amplifying cells promotes epithelial cell proliferation and lengthens crypts. These findings suggest that SBR reduces p85α and TP53, which increases survivin and intestinal epithelial cell expansion during therapeutic adaptation in patients with short bowel syndrome. PMID:27157990

  2. Expression of survivin in human gastric carcinoma and gastric carcinoma model of rats

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    Xiao-Dong Zhu; Geng-Jin Lin; Li-Ping Qian; Zhong-Qing Chen

    2003-01-01

    AIM: To study the expression of survivin, an inhibitor of apoptosis protein, in human gastric carcinomas and gastric carcinoma models of rats.METHODS: With the method of immunohistochemical staining, we studied the expression of survivin in 20 cases of chronic gastritis and 56 cases of gastric carcinomas. We used N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and high dose sodium-chloride diet to induce rat gastric carcinomas. Survivin expression was studied in glandular stomachs of normal rats, adenocarcinomas and tissues adjacent to the tumor, as well as in rats during the induction period.RESULTS: Survivin was expressed in 27 of 56 (48.2 %)cases of human gastric carcinoma tissues and 1 of 20 (5 %)cases of chronic gastritis. It was found that the expression of survivin had no relation with the elements of age, tumor depth, tumor size, and disease stage, but was significantly related to histological type. The positive rate of survivin expression in cases of intestinal type was significantly higher than that in cases of diffuse type (P<0.05). In animal experiments, survivin expression in glandular stomachs of normal rats, of rats in middle induction period, in adenocarcinomas and tissues adjacent to tumor were 0,40.0 %, 78.3 % and 38.9 %, respectively. Compared with the survivin expression in normal rats, the differences were significant.CONCLUSION: These data imply that survivin plays an important role in the onset of gastric carcinoma and that high survivin expression is an early event of gastric carcinoma.

  3. Relationship between survivin expression and recurrence, and prognosis in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To study the expression of the inhibitor of apoptosis protein survivin in hepatocellular carcinoma (HCC), and its correlation with clinicopathological factors, cell proliferation, recurrence and prognosis after hepatectomy.METHODS: Immunohistochemical staining of survivin and Ki-67 was performed by the standard streptavidinperoxidase technique on paraffin sections of 55 cases of HCC.RESULTS: The positive rate of survivin in HCC was 52.7% (29/55). Significant correlation was found between survivin expression with portal vein thrombi and intrahepatic matastasistic nodes (P < 0.05). The recurrent rate in survivin-positive HCC was significantly higher than that in survivin-negative HCC after hepatectomy, the 1- and 3-year survival rate in patients with survivin-positive tumors was significantly lower than that in patients with survivin-negative tumors (58.62 and 10.34% vs 76.92 and 30.77%, P < 0.05, log-rank test).The proliferation index (Ki-67) in survivin-positive HCC (33.83% ± 18.90%) was significantly higher than that in survivin-negative HCC (19.60% ± 19.35%) (P < 0.05).CONCLLSION: Survivin may play an important role in progression of HCC by promoting cell proliferation, and may be positively correlated with high risk of disease recurrence and poor prognosis in HCC. Its expression may serve as a prognostic factor for patients with HCC after hepatectomy.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Institute of Scientific and Technical Information of China (English)

    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

  6. Detection of Survivin and COX-2 in Thyroid Carcinoma: Anaplastic Carcinoma Shows Overexpression of Nuclear Survivin and Low COX-2 Expression

    OpenAIRE

    Kim, Young A; Chang, Meesoo; Park, Young Joo; Kim, Ji Eun

    2012-01-01

    Background Overexpression of survivin, a member of the inhibitors of apoptosis protein, has been reported in various carcinomas, and its interaction with cyclooxygenase 2 (COX-2) results in accelerated tumor progression. The purpose of this study is to investigate the immunohistochemical expression of survivin and COX-2 in benign and malignant thyroid tissues and to define its association with pathologic and clinical features. Methods We examined expression of survivin and COX-2 by immunohist...

  7. Correlation between survivin mRNA expression and homoharringtonine induced apoptosis of malignant hematopoietic cells

    Institute of Scientific and Technical Information of China (English)

    CAI Zhen; BAO Han-ying; LIN Mao-fang

    2005-01-01

    Background The inhibitor of apoptosis (IAP) gene family is involved in the suppression of apoptotic cell death as well as an increasing number of seemingly unrelated cellular functions. It is not known, however, whether IAP expression in malignant hematopoietic cells is affected by chemotherapeutic agents such as homoharringtonine (HHT). In this study, we investigated mRNA expression levels of IAPs, especially survivin, in various hematopoietic cell lines in relation with apoptosis induced by HHT. Methods Semiquantitative reverse transcriptase polymerase chain reaction was used to determine survivin mRNA levels. Cell apoptosis was examined by flow cytometry. Cell viability and proliferation assay was evaluated by MTT. The experiments were performed on the malignant hematopoietic cell lines MUTZ-1, K562, Jurkat, RMPI and HL60, with or without survivin antisense-oligodeoxynucleotides (AS-ODN) and HHT.Results The expression levels of survivin mRNA were variable in the cell lines and negatively correlated to HHT induced cell apoptosis. Survivin AS-ODN significantly decreased mRNA level of survivin, but not those of bax and bcl-2. Survivin also inhibited MUTZ-1 cell growth and induced apoptosis in a dose dependent manner. AS-ODN and HHT showed synergistic effect on MUTZ-1 cell growth.Conclusion The apoptotic effect of HHT on the hematopoietic cell lines is associated with decreased level of survivin expression. Survivin could be a new marker for drug sensitivity and a new target for cancer treatment.

  8. 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.

  9. SURVIVIN AND TUMOR

    Institute of Scientific and Technical Information of China (English)

    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.

  10. 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.

  11. Immunohistochemical Expression of Survivin in Breast Carcinoma: Relationship with Clinico pathological Parameters, Proliferation and Molecular Classification

    International Nuclear Information System (INIS)

    Background and Objective: Survivin is a novel member of the inhibitor of apoptosis (IAP) gene family. It is associated with more aggressive behavior and parameters of poor prognosis in most human cancers including gastric, colorectal and bladder carcinomas. However, conflicting data exist on its prognostic effect in breast cancer. This current study is designed to assess survivin expression in breast carcinoma relating results with clinico pathological parameters, proliferation (MIB-1) and molecular classification. Material and Methods: Our retrospective study com- prised of 65 archived cases of breast carcinoma. Samples from the tumor and the adjacent normal breast tissue were immuno stained for survivin and MIB-1. Nuclear and cytoplasmic survivin expression was evaluated in normal breast tissue and carcinoma regarding both the intensity and the percentage of positive cells. ER, PR, HER2 were used as surrogate markers to classify the cases into four molecular subtypes. Results: Survivin expression was detected in 78.5% of breast carcinomas. The adjacent normal breast tissue was immuno negative. Survivin expression showed significant association with increased tumor size ( p <0.0001), high histologic grade ( p =0.04), lymph node metastases ( p <0.001), advanced tumor stage ( p <0.0001), MIB-1 expression ( p =0.02), negative estrogen receptor status ( p =0.01) and negative progesterone receptor status ( p <0.0001). The subcellular localization of survivin significantly related to histologic grade, stage and lymph node involvement. The percentage of TNP (triple negative phenotype) and HER2+/ER-PR- tumors expressing survivin were significantly higher compared to the Luminal subtypes ( p =0.01). Conclusion: Survivin expression was associated with parameters of poor prognosis in breast cancer. Moreover, the cancer-specific expression of survivin, coupled with its importance in inhibiting cell death and in regulating cell division, makes it a potential target for novel

  12. C-Reactive Protein Inhibits Survivin Expression via Akt/mTOR Pathway Downregulation by PTEN Expression in Cardiac Myocytes

    OpenAIRE

    Beom Seob Lee; Soo Hyuk Kim; Jaewon Oh; Taewon Jin; Eun Young Choi; Sungha Park; Sang-Hak Lee; Ji Hyung Chung; Seok-Min Kang

    2014-01-01

    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 ...

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. Knockdown of Survivin Expression by siRNA Induces Apoptosis of Hepatocellular Carcinoma Cells

    Institute of Scientific and Technical Information of China (English)

    WU Liang; WANG Ying; TIAN Deying

    2007-01-01

    Survivin, a newly identified member of IAP family, is a powerful apoptosis-inh ibiting factor. It is expressed in embryonic tissues as well as in the majority of human cancers, but not in most normal adult tissues. The cancer-specific expression of survivin makes it a potential target for cancer treatment. A survivin-specific small inhibitory RNA (siRNA) was introduced into hepatocellular carcinoma cells to investigate its effect on cancer cell apoptosis, growth and sensitivity to chemotherapeutic drugs. It was found that expressions of survivin protein and proliferation index (PI) in siRNA groups were significantly decreased, the apoptosis index (AI) of siRNA groups was significantly higher than those of others groups, and the growth inhibition rate (GIR) of chemotherapeutic drugs in siRNA groups were significantly higher than those of other groups. Our study suggests that the expression of survivin may be significantly decreased in hepG2 cell after siRNA transfection.siRNA targeting survivin could induce cell apoptosis, inhibit cell proliferation and sensitize hepatocarcinoma cells to chemotherapy. Our findings provide preliminary evidence for the therapeutic use of survivin-targeted RNA interference for human tumors that express high levels of this molecule.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. Inhibition of survivin expression and mechanisms of reversing drug-resistance of human lung adenocarcinoma cells by siRNA

    Institute of Scientific and Technical Information of China (English)

    LIU Jing-lei; WANG Yan; JIANG Ji; KONG Rui; YANG Yan-mei; JI Hong-fei; SHI Yu-zhi

    2010-01-01

    Background Survivin, a member of the inhibitor of apoptosis protein (IAP) family, overexpresses in tumor cells and not expresses in terminally differentiated adult tissues. This study aimed to investigate the effects of survivin-specific siRNA on cell proliferation, apoptosis and chemosensitivity to cisplatin in vitro and in vivo and explore the mechanisms about decreasing expression of survivin in reversing cancer cells resistance to chemotherapeutic drug.Methods Survivin-specific siRNA was transfected into A549/DDP cells. The expression of survivin and lung resistance-related protein (LRP) mRNA levels were determined by RT-PCR, chemosensitivity of A549/DDP (cisplatin)cells to cisplatin was determined by MTT assay, and apoptosis and cell cycle were determined by flow cytometry (FCM).The protein expression levels of survivin, LRP, cyclin-D1, caspase-3 and bcl-2 were determined by Western blotting analyses. The effect of survivin siRNA inhibition on tumor growth was studied in athymic nude mice in vivo.Results Survivin-specific siRNA efficiently down-regulated survivin expression. The cell cycle was arrested at G2/M phase, and apoptosis was obviously found. Inhibition of survivin expression could make the IC50 and drug-resistant index of cisplatin decrease, and enhance the cancer cells sensitivity to cisplatin. After transfection by survivin-specific siRNA, expression of LRP and cyclin-D1 were downregulated, caspase-3 expression was upregulated, bcl-2 expression had no obvious change. The animal experiment confirmed knockdown of survivin could inhibit the tumor growth.Conclusions Survivin-specific siRNA can efficiently suppress the expression of survivin, increase apoptosis, inhibit cells proliferation and enhance the chemosensitivity to cisplatin in vitro and in vivo. Suppression of survivin expression helping to reverse drug-resistance may have relationship with downregulation of LRP and upregulation of caspase-3.Anti-tumor strategies based on the inhibition of

  6. 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.

  7. 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

  8. Expression of a novel apoptosis inhibitor-survivin in colorectal carcinoma

    Institute of Scientific and Technical Information of China (English)

    Hai-Yan Tan; Jun Liu; Shan-Min Wu; He-Sheng Luo

    2005-01-01

    AIM: To investigate the role of survivin expression in the pathogenesis of colorectal carcinoma.METHODS: Immunohistochemistry S-P method and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were used to detect the expression of survivin and apoptotic cell in situ in colorectal cancerous tissues, para-cancerous tissues and normal tissues of 48 cases of colorectal carcinoma.RESULTS: The survivin positive unit (PU) was higher in cancerous tissues (38.76±5.14)than in para-cancerous (25.17±7.26) or normal tissues (0.57±0.03) (P<0.05).The apoptosis index (AI) of para-cancerous tissues was(7.51±2.63%) higher than cancerous tissues (4.65±1.76%).The expression of survivin was associated with pathological grade, lymph node metastasis and Dukes stage of colorectal carcinoma.CONCLUSION: Survivin expression may play an important role in carcinogenesis of colorectal carcinoma and may be associated with malignant biological behaviors of colorectal carcinoma.

  9. 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...

  10. Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity

    Institute of Scientific and Technical Information of China (English)

    Hai-Tao Guan; Xing-Huan Xue; Zhi-Jun Dai; Xi-Jing Wang; Ang Li; Zhao-Yin Qin

    2006-01-01

    AIM: To investigate the inhibitory effect of small interfering RNA (siRNA) on the expression of survivin in pancreatic cancer cell line PC-2 and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity.METHODS: A siRNA plasmid expression vector against survivin was constructed and transfected into PC-2 cells with LipofectamineTM 2000. The down regulation of survivin expression was detected by semi-quantitive RT-PCR and immunohistochemical SP method and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity was detected by flow cytometry.RESULTS: The sequence-specific siRNA efficiently and specifically down-regulated the expression of survivin at both mRNA and protein levels. The expression inhibition ratio was 81.25% at mRNA level detected by semiquantitive RT-PCR and 74.24% at protein level detected by immunohistochemical method. Forty-eight hours after transfection,apoptosis was induced in 7.03% cells by siRNA and in 14.58% cells by siRNA combined with radiation.CONCLUSION: The siRNA plasmid expression vector against survivin can inhibit the expression of survivin in PC-2 cells efficiently and specifically. Inhibiting the expression of survivin can induce apoptosis of PC-2 cells and enhance its radiosensitivity significantly. RNAi against survivin is of potential value in gene tnerapy of pancreatic cancer.

  11. 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.

  12. 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

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    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.

  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. The antiapoptotic gene survivin is highly expressed in human chondrosarcoma and promotes drug resistance in chondrosarcoma cells in vitro

    International Nuclear Information System (INIS)

    Chondrosarcoma is virtually resistant to chemotherapy and radiation therapy. Survivin, the smallest member of the inhibitor of apoptosis protein family, is a critical factor for tumor progression and resistance to conventional therapeutic approaches in a wide range of malignancies. However, the role of survivin in chondrosarcoma has not been well studied. We examined the importance of survivin gene expression in chondrosarcoma and analysed its influences on proliferation, apoptosis and resistance to chemotherapy in vitro. Resected chondrosarcoma specimens from which paraffin-embedded tissues could be extracted were available from 12 patients. In vitro experiments were performed in human chondrosarcoma cell lines SW1353 and Hs819.T. Immunohistochemistry, immunoblot, quantitative PCR, RNA interference, gene-overexpression and analyses of cell proliferation and apoptosis were performed. Expression of survivin protein was detected in all chondrosarcoma specimens analyzed, while undetectable in adult human cartilage. RNA interference targeting survivin resulted in a G2/M-arrest of the cell cycle and led to increased rates of apoptosis in chondrosarcoma cells in vitro. Overexpression of survivin resulted in pronounced resistance to doxorubicin treatment. These findings indicate that survivin plays a role in the pathogenesis and pronounced chemoresistance of high grade chondrosarcoma. Survivin antagonizing therapeutic strategies may lead to new treatment options in unresectable and metastasized chondrosarcoma

  15. Downregulation of survivin expression exerts antitumoral effects on mouse breast cancer cells in vitro and in vivo

    Science.gov (United States)

    MA, WEN-HUI; LIU, YONG-CHAO; XUE, MEI-LAN; ZHENG, ZHENG; GE, YIN-LIN

    2016-01-01

    Metastasis constantly occurs in the majority of cases of primary breast cancer at late stage or following surgical treatment. Survivin, a member of the inhibitor of apoptosis protein family, has long been recognized as a promising anticancer target, but its antitumor effects remain largely unexplored. In order to elucidate the role of survivin in breast cancer metastasis, short interfering RNA (siRNA) was used in the present study to specifically downregulate survivin expression in the murine breast cancer cell line 4T1. The results demonstrated that blocking the expression of survivin by siRNA inhibited the proliferation, migration and invasion abilities of murine breast cancer cells in vitro. Vascular endothelial growth factor (VEGF)-C is a lymphatic endothelial cell-stimulating factor that may lead to the formation of lymphatic vessels in lymph nodes. In the present study, the inhibition of survivin by siRNA was able to reduce the overexpression of VEGF-C in 4T1 cells. Furthermore, intratumoral injections of the survivin-siRNA significantly inhibited the growth of orthotopically transplanted 4T1 tumors in vivo. In addition, the number of pulmonary metastases and the microlymphatic vessel density were significantly reduced in vivo, following transfection with survivin-siRNA. The results of the present study suggested that the Akt/hypoxia-inducible factor-1α signaling pathway participates in the survivin-mediated downregulation of VEGF-C expression observed in breast cancer cells treated with survivin-siRNA. Therefore, the use of siRNA specifically targeting survivin may be a potential anticancer method in the future. PMID:26870183

  16. Survivin, a novel target of the Hedgehog/GLI signaling pathway in human tumor cells.

    Science.gov (United States)

    Vlčková, K; Ondrušová, L; Vachtenheim, J; Réda, J; Dundr, P; Zadinová, M; Žáková, P; Poučková, P

    2016-01-01

    Survivin, an important antiapoptotic protein, is expressed in tumors, whereas in normal tissues the expression of this protein is extremely low, defining a role for survivin as a cancer gene. Survivin exhibits multifunctional activity in tumor cells. However, why survivin expression is sharply and invariably restricted to tumor tissue remains unclear. Here, we identified 11 putative consensus binding sites for GLI transcription factors in the survivin promoter and characterized the promoter activity. Inhibitors of the Hedgehog/GLI pathway, cyclopamine and GANT61, decreased the promoter activity in reporter assays. ΔNGLI2 (which lacks the repressor domain) was the most potent vector in activating the survivin promoter-reporter. Moreover, GANT61, a GLI1/2 inhibitor, repressed endogenous survivin protein and mRNA expression in most cells across a large panel of tumor cell lines. Chromatin immunoprecipitation showed GLI2 binding to the survivin promoter. The ectopic GLI2-evoked expression of endogenous survivin was observed in normal human fibroblasts. GANT61 decreased survivin level in nude mice tumors, mimicking the activity of GANT61 in cultured cells. The immunohistochemistry and double immunofluorescence of human tumors revealed a correlation between the tissue regions showing high GLI2 and survivin positivity. Thus, these results demonstrated that survivin is a classical transcriptional target of GLI2, a Hedgehog pathway signaling effector. This potentially reflects the high expression of survivin in human tumor cells. As the Hedgehog pathway is upregulated in virtually all types of cancer cells, these findings substantially contribute to the explanation of uniform survivin expression in tumors as a potential target for the development of a more effective treatment of cancers through the inhibition of GLI2 to restrain survivin activity. PMID:26775700

  17. 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.

  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. 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.

  1. 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.

  2. 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.

  3. Serum Survivin Increases in Prolactinoma

    OpenAIRE

    Dellal, Fatma Dilek; Niyazoglu, Mutlu; Gorar, Suheyla; Ademoglu, Esranur; Candan, Zehra; Bekdemir, Handan; Hacioglu, Yalcin; Kaya, Fatih Oner

    2015-01-01

    Background Prolactinoma is the most common adult pituitary adenoma. Survivin is a member of the family of inhibitors of apoptosis proteins. Its expression is observed in many tumors. Survivin expression has shown in prolactinoma tissue before but no study exists showing serum survivin level. The aim of the present study was to investigate serum survivin levels in patients with prolactinoma and demonstrate its value in diagnosis of the disease. Methods The group of patients consisted of 25 wom...

  4. Expression of survivin detected by immunohistochemistry in the cytoplasm and in the nucleus is associated with prognosis of leiomyosarcoma and synovial sarcoma patients

    International Nuclear Information System (INIS)

    Survivin, a member of the inhibitor of apoptosis-protein family suppresses apoptosis and regulates cell division. It is strongly overexpressed in the vast majority of cancers. We were interested if survivin detected by immunohistochemistry has prognostic relevance especially for patients of the two soft tissue sarcoma entities leiomyosarcoma and synovial sarcoma. Tumors of leiomyosarcoma (n = 24) and synovial sarcoma patients (n = 26) were investigated for their expression of survivin by immunohistochemistry. Survivin expression was assessed in the cytoplasm and the nucleus of tumor cells using an immunoreactive scoring system (IRS). We detected a survivin expression (IRS > 2) in the cytoplasm of 20 leiomyosarcomas and 22 synovial sarcomas and in the nucleus of 12 leiomyosarcomas and 9 synovial sarcomas, respectively. There was no significant difference between leiomyosarcoma and synovial sarcoma samples in their cytoplasmic or nuclear expression of survivin. Next, all sarcoma patients were separated in four groups according to their survivin expression in the cytoplasm and in the nucleus: group 1: negative (IRS 0 to 2); group 2: weak (IRS 3 to 4); group 3: moderate (IRS 6 to 8); group 4: strong (IRS 9 to 12). In a multivariate Cox's regression hazard analysis survivin expression detected in the cytoplasm or in the nucleus was significantly associated with overall survival of patients in group 3 (RR = 5.7; P = 0.004 and RR = 5.7; P = 0.022, respectively) compared to group 2 (reference). Patients whose tumors showed both a moderate/strong expression of survivin in the cytoplasm and a moderate expression of survivin in the nucleus (in both compartments IRS ≥ 6) possessed a 24.8-fold increased risk of tumor-related death (P = 0.003) compared to patients with a weak expression of survivin both in the cytoplasm and in the nucleus. Survivin protein expression in the cytoplasma and in the nucleus detected by immunohistochemistry is significantly associated with

  5. 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.

  6. 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.

  7. Survivin inhibits anti-growth effect of p53 activated by aurora B

    International Nuclear Information System (INIS)

    Genomic instability and apoptosis evasion are hallmarks of cancer, but the molecular mechanisms governing these processes remain elusive. Here, we found that survivin, a member of the apoptosis-inhibiting gene family, and aurora B kinase, a chromosomal passenger protein, were co-overexpressed in the various glioblastoma cell lines and tumors. Notably, exogenous introduction of the aurora B in human BJ cells was shown to decrease cell growth and increase the senescence-associated β-galactosidase activity by activation of p53 tumor suppressor. However, aurora B overexpression failed to inhibit cell proliferation in BJ and U87MG cells transduced with dominant-negative p53 as well as in p53-/- mouse astrocytes. Aurora B was shown to increase centrosome amplification in the p53-/- astrocytes. Survivin was shown to induce anchorage-independent growth and inhibit anti-proliferation and drug-sensitive apoptosis caused by aurora B. Overexpression of both survivin and aurora B further accelerated the proliferation of BJ cells. Taken together, the present study indicates that survivin should accelerate tumorigenesis by inhibiting the anti-proliferative effect of p53 tumor suppressor that is activated by aurora B in normal and glioblastoma cells containing intact p53

  8. The Mechanism of Radiosensitization by YM155, a Novel Small Molecule Inhibitor of Survivin Expression, is Associated with DNA Damage Repair

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    Songliu Hu

    2015-09-01

    Full Text Available Background/Aims: Survivin, a member of the inhibitor of apoptosis protein family, is an attractive target for cancer therapy. We investigated the effects of YM155, a small molecule inhibitor of survivin expression, on the radiosensitivity of human non-small cell lung cancer (NSCLC cell lines and elucidated a relationship between the cellular localization of survivin and DNA double-strand break repair. Methods: The cellular distribution of survivin was determined by Western blotting of subcellular fractions and by immunofluorescent staining in A549 NSCLC cells. Radiation-induced DNA damage was evaluated based on histone H2AX phosphorylation and foci formation. The relationship between the cellular localization of survivin and DNA double-strand break repair was analyzed by Western blotting and co-immunoprecipitations. Results: YM155 down-regulated survivin expression in NSCLC cells in a concentration- and time-dependent manner. An in vitro clonogenic survival assay revealed that YM155 increased the sensitivity of NSCLC cells to radiation. After irradiation, we observed a rapid accumulation of survivin in the nucleus. An immunofluorescent analysis of histone γ-H2AX demonstrated that the inhibition of survivin expression by YM155 resulted in impaired DNA double-strand break repair. Co-immunoprecipitation assays using nuclear extracts revealed an interaction between survivin, Ku70, γ-H2AX, and DNA-PKcs. Furthermore, S2056 autophosphorylation of DNA-PKcs was reduced in survivin-depleted cells. Conclusions: These results suggested that YM155 sensitized NSCLC cells to radiation, at least in part by inhibiting DNA repair and enhancing apoptosis via the down-regulation of survivin expression. YM155 pretreatment inhibited DNA-PKcs autophosphorylation at S2056. Nuclear survivin was involved in DNA double-strand break repair via interactions with members of the DNA double-strand break repair machinery.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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

    Directory of Open Access Journals (Sweden)

    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.

  11. Expression of Survivin in Patients with acute myeloid Leukemia and effect of amifostine on survivin expression%骨髓细胞中 survivin 表达与急性髓系白血病的关系及阿米福汀对其的影响

    Institute of Scientific and Technical Information of China (English)

    杨颖莹; 刘郝静; 王丹

    2015-01-01

    目的:分析骨髓细胞中 survivi 表达与进行髓系白血病的关系及阿米福汀对其的影响。方法选取我院在2013年2月-2015年4月初诊为急性髓系白血病患者60例为观察组,另外选取我院同期健康体检人员50例为对照组,分析骨髓细胞 survivin 表达,并分析阿米福汀对 survivin 表达的影响。结果观察组患者 survivin 表达阳性率(83.3%)明显高于对照组(32.0%),P <0.05,不同浓度阿米福汀均能够抑制 HL-60细胞,不同干预时间组与丁组之间差异明显,P <0.05。疗效评估,37例患者白血病细胞 survivin 表达阳性,9例患者表达阴性,存在明显差异。不同阿米福汀浓度作用下,survivin 表达灰度值均与丁组存在明显差异,P >0.05,不同干预时间作用下 survivin 表达存在明显差异,P <0.05。结论骨髓细胞中 survivin 表达与白血病发展存在密切关系,阿米福汀能够下调凋亡基因 sur-vivin 的表达。%Objective To expression of Survivin in Patients with acute myeloid Leukemia and explore the effect of amifostine on survivin.Methods 60 patients with Leukemia in hospital as observer Group were divided from february 2013 to april 2015,healthy patients in our hospital over the same period were selected as the control group,survivin ex-pression in bone marrow cells were analyzed,the impact of amifostine on survivin expression was analyzed.Results The positive rate of survivin expression in observation group (83.3%)was significantly higher (32.0%),P 0.05,the next time the role of different inter-ventions survivin expression significantly different,P <0.05.Conclusion Closely relationship between survivin expres-sion in myeloid leukemia cells,amifostine can decrease the expression of apoptotic gene survivin.

  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

    Institute of Scientific and Technical Information of China (English)

    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

    Institute of Scientific and Technical Information of China (English)

    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. Expression of HER-2 in MCF-7 breast cancer cells modulates anti-apoptotic proteins Survivin and Bcl-2 via the extracellular signal-related kinase (ERK) and phosphoinositide-3 kinase (PI3K) signalling pathways

    International Nuclear Information System (INIS)

    The oncoprotein HER-2 is over-expressed and/or has undergone gene amplification in between 20 to 30% of breast and ovarian cancers. HER-2 amplified breast cancer is associated with a poor prognosis and increased resistance to chemo- and hormonal therapy. Data supporting the transforming potential of HER-2 are irrefutable but the mechanism by which HER-2 contributes to this process is complex and a unified model of HER2-induced increased cell proliferation and survival has not emerged. To understand the initial event(s) that take place by HER-2 over expression, we studied the effect of short term induction of HER-2 expression in the MCF7 breast cancer cell line. We examined the modulation of apoptotic pathways by tetracycline-regulated HER-2 expression for 48 hrs in the MCF7 breast cancer cell line. Specific inhibitors were used to determine signalling pathways that are required for HER-2 induced up-regulation of survivin. Tetracycline regulated short term over expression of HER-2 in the MCF7 cell line increased the antiapoptotic proteins Bcl-2 and survivin levels. Significant increase of extracellular signal-related kinase (ERK) activation but not AKT1, AKT2 and STAT3 was observed in HER-2 over-expressing MCF7 cells. Specific inhibitors of ERK, and phosphoinositide-3 kinase (PI3K), inhibited the HER-2 induced up-regulation of survivin. We did not observe a change in survivin and NF-κB promoter activity in HER-2 expressing MCF7 cells. Our results indicate that short term over expression of HER-2 up regulates antiapoptotic proteins Bcl-2 and survivin in MCF7 cells. We determined that survivin is up-regulated via ERK activation and PI3K signalling. Additionally we show that survivin up-regulation is not at transcriptional level. These data provide insight into the mechanism(s) by which induction of HER-2 over expression up-regulates survivin and Bcl-2 and identifies new targets for therapy of breast cancer

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

    Science.gov (United States)

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. The expression of antiapoptotic protein survivin is transcriptionally upregulated by DEC1 primarily through multiple sp1 binding sites in the proximal promoter

    OpenAIRE

    Li, Y.; Xie, M.; Yang, J; D. Yang; Deng, R.; Wan, Y; B. Yan

    2006-01-01

    Human differentially expressed in chondrocytes (DEC), mouse stimulated with retinoic acid and rat split and hairy related proteins constitute a structurally distinct class of the basic helix-loop-helix proteins. DEC1is abundantly expressed in tumors and protects against apoptosis induced by serum starvation. In this study, we report that DEC1 antiapoptosis is achieved by inducing survivin, an antiapoptotic protein. In paired tumor–normal tissues, survivin and DEC1 exhibited a paralleled expre...

  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. Comparison of Immunohistochemical Expression of Antiapoptotic Protein Survivin in Normal Oral Mucosa, Oral Leukoplakia, and Oral Squamous Cell Carcinoma

    OpenAIRE

    Amita Negi; Abhiney Puri; Rakhi Gupta; Rajat Nangia; Alisha Sachdeva; Megha Mittal

    2015-01-01

    Background. Oral squamous cell carcinoma is the sixth most frequent malignant tumor worldwide and the third most common cancers in developing countries. Oral leukoplakia is the best-known precursor lesion of oral squamous cell carcinoma. The aim of the present study was to compare immunohistochemical expression of antiapoptotic protein survivin in normal oral mucosa, oral leukoplakia, and oral squamous cell carcinoma. Method. Total 45 specimens of formalin fixed paraffin embedded tissue block...

  8. Survivin Antisense Oligodeoxy-Nucleotid Induces Apoptosis in Leukaemia Cell Line K562

    Institute of Scientific and Technical Information of China (English)

    Lijun Chen; Qiuyue Jin; Hong Xie; Ruimin Wang; Li Yao

    2006-01-01

    OBJECTIVE To investigate the effects of survivin antisense oligodeoxynucleotid (ASODN) on proliferation and apoptosis in the chronic myeloid leukemia cell line K562.METHODS Different concentrations of an antisense oligodeoxy-nucleotid and control sequence (scrambled ODN) targeting the survivin gene were transferred into K562 by a lipofectin reagent. The MTT assay was used to measure the growth inhibitory rate, IC50, and to observe the cytotoxicity of survivin ASODN in the K562 cells. The morphologic changes in the nucleus and the apoptotic rate were observed by Hoechst33342/PI staining.Caspase-3 activity was evaluated by a kinase activity assay. The changes of survivin protein expression after transfection were detected by Western blots.RESULTS Eight hours after transfection, fluorescence in the K562 cells was well distributed. Treatment of the cells for 44 h with different concentrations of survivin ASODN produced a IC50 of 800 nmol/L. The growth inhibitory rate with 200, 400, 600 and 1000 nmol/L of survivin ASODN was 15.8±1.6%, 23.8±5.9%, 37.1±5.6% and 77.3±2.5% respectively. After 36 h of of survivin ASODN treatment, distinct morphologic changes characteristic of cell apoptosis such as karyopyknosis and conglomeration were observed by Hoechst33342/PI staining. Caspase-3 activity increased significantly after treatment of the cells with different concentrations of survivin ASODN (P<0.01)and following treatment with 800 nmol/L survivin ASODN, survivin expression decreased significantly.CONCLUSION Survivin ASODN exerts an anti-cancer effect by inducing apoptosis in K562 leukaemia cells. Up-regulated expression of caspase3 may play a role in this process.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. 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. PMID:24395429

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

    International Nuclear Information System (INIS)

    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

  12. Curcumin reduces the expression of survivin, leading to enhancement of arsenic trioxide-induced apoptosis in myelodysplastic syndrome and leukemia stem-like cells.

    Science.gov (United States)

    Zeng, Yingjian; Weng, Guangyang; Fan, Jiaxin; Li, Zhangqiu; Wu, Jianwei; Li, Yuanming; Zheng, Rong; Xia, Pingfang; Guo, Kunyuan

    2016-09-01

    Low response, treatment-related complications and relapse due to the low sensitivity of myelodysplastic syndrome (MDS) and leukemia stem cells (LSCs) or pre‑LSCs to arsenic trioxide (ATO), represent the main problems following treatment with ATO alone in patients with MDS. To solve these problems, a chemosensitization agent can be applied to increase the susceptibility of these cells to ATO. Curcumin (CUR), which possesses a wide range of anticancer activities, is a commonly used chemosensitization agent for various types of tumors, including hematopoietic malignancies. In the present study, we investigated the cytotoxic effects and potential mechanisms in MDS-SKM-1 and leukemia stem-like KG1a cells treated with CUR and ATO alone or in combination. CUR and ATO exhibited growth inhibition detected by MTT assays and apoptosis analyzed by Annexin V/PI analyses in both SKM-1 and KG1a cells. Apoptosis of SKM-1 and KG1a cells determined by Annexin V/PI was significantly enhanced in the combination groups compared with the groups treated with either agent alone. Further evaluation was performed by western blotting for two hallmark markers of apoptosis, caspase-3 and cleaved-PARP. Co-treatment of the cells with CUR and ATO resulted in significant synergistic effects. In SKM-1 and KG1a cells, 31 and 13 proteins analyzed by protein array assays were modulated, respectively. Notably, survivin protein expression levels were downregulated in both cell lines treated with CUR alone and in combination with ATO, particularly in the latter case. Susceptibility to apoptosis was significantly increased in SKM-1 and KG1a cells treated with siRNA-survivin and ATO. These results suggested that CUR increased the sensitivity of SKM-1 and KG1a cells to ATO by downregulating the expression of survivin. PMID:27430728

  13. Expression of antiapoptosis gene survivin in luteinized ovarian granulosa cells of women undergoing IVF or ICSI and embryo transfer: clinical correlations

    Directory of Open Access Journals (Sweden)

    Varras Michail

    2012-09-01

    Full Text Available Abstract Background The purpose of the study was to determine the incidence of survivin gene expression in human granulosa cells during ovarian stimulation in Greek women with normal FSH levels, undergoing IVF or ICSI and to discover any correlation between levels of gene expression and clinical parameters, efficacy of ovulation or outcomes of assisted reproduction. Methods Twenty nine women underwent ovulation induction for IVF or ICSI and ET with standard GnRH analogue-recombinant FSH protocol. Infertility causes were male and tubal factor. Cumulus–mature oocyte complexes were denuded and the granulosa cells were analyzed for each patient separately using quantitative reverse transcription polymerase chain reaction analysis for survivin gene expression with internal standard the ABL gene. Results The ABL and survivin mRNA were detected in granulosa cells in 93.1%. The expression levels of survivin were significantly lower in normal women (male infertility factor compared to women with tubal infertility factor (p = 0.007. There was no additional statistically significant correlation between levels of survivin expression and estradiol levels or dosage of FSH for ovulation induction or number of dominant follicles aspirated or number of retrieved oocytes or embryo grade or clinical pregnancy rates respectively. Conclusions High levels of survivin mRNA expression in luteinized granulosa cells in cases with tubal infertility seem to protect ovaries from follicular apoptosis. A subpopulation of patients with low levels of survivin mRNA in granulosa cells might benefit with ICSI treatment to bypass possible natural barriers of sperm-oocyte interactions.

  14. 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

  15. 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...

  16. 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.

  17. Survivin in survival of hepatocellular carcinoma.

    Science.gov (United States)

    Su, Changqing

    2016-09-01

    Survivin is an anti-apoptotic protein belonging to the inhibitor of apoptosis protein (IAP) family. It is involved in the regulation of important physiological and pathological processes in cells and functions to inhibit cell apoptosis and promote cell proliferation. Normally and terminally differentiated tissues are nearly negative for survivin. In contrast, survivin is highly expressed in most human tumor tissues, including hepatocellular carcinoma (HCC). The abnormal overexpression of survivin is closely related to the malignant biological behaviors of tumors. During the development and progression of HCC, the high level of survivin expression promotes cancer cell proliferation, inhibits cancer cell apoptosis, induces tumor stromal angiogenesis, reduces the sensitivity of cancer cells to radiotherapy and chemotherapy, and ultimately affects the prognosis of patients with HCC. Survivin expression is regulated by a large number of factors. The latest discovery indicated that the transcription factor octamer-binding transcription factor 4 (OCT4) enhances the expression of survivin though cyclin D1 (CCND1), which, in part, accounts for tumor cell proliferation, recurrence and metastasis. Survivin plays key roles in HCC, which renders it an ideal target for the treatment of HCC. The present article reviews the research progress on the relationship between survivin and HCC and on the HCC treatment strategies targeting survivin. PMID:26118774

  18. 宣威昆明两地肺腺癌患者中生存素表达的比较研究%The expression of Survivin in lung adenocarcinoma:a comparative study in Xuanwei and Kunming

    Institute of Scientific and Technical Information of China (English)

    Weiwei Wang; Gaofeng Li; Zhipeng Hong; Shaojia Wang; Ruibing Cheng; Shoujun Deng; Jupeng Zhang

    2010-01-01

    Objective: The aim of our study was to investigate the expression of Survivin in lung adenocarcinoma of Xuanwei and Kunming patients.Methods: Twenty-five specimens of Xuanwei patients and 28 specimens of Kunming patients were observed and analyzed.The Survivin expression was detected by immunohistochemistry.The results were quantitatively analyzed by image analysis system.Results: There were significant differences in Survivin expression (P<0.01) between Xuanwei patients and Kunming patients.Conclusion: Survivin expression in lung adenocarcinoma of Xuanwei patients was significantly higher than that of Kunming patients.The pathogenesis of lung adenocarcinoma might be different between Xuanwei patients and Kunming patients.High Survivin expression might be one of the reasons to explain high incidence of lung cancer in Xuanwei.

  19. Correlations in survivin expression with the expression of p53 and bcl-2 in invasive ductal carcinoma of the breast.

    Science.gov (United States)

    Al-Joudi, F S; Iskandar, Z A; Imran, A K

    2007-09-01

    This work studied the correlations between survivin, bcl-2 and p53 in infiltrating ductal carcinoma of the breast. A total number of 382 cases were collected from 3 hospitals in northeastern Malaysia. Survivin, bcl-2 and p53 were detected by immunohistochemistry on samples prepared from tissue blocks. Significant correlations were found between tumor histological grades and tumor size and lymph node involvement. Highly significant statistical correlations (pfashion, implying that many of these cases may share common abnormalities. PMID:18041310

  20. 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.

  1. Latent membrane protein 1 inhibits apoptosis induced by 60 irradiation via Survivin triggering signal-pathway

    International Nuclear Information System (INIS)

    Objective: To investigate the anti-apoptosis mechanism of EB virus encoden latent membrane protein 1 (LMP1) via the survivin signal transduction pathway after irradiation induction. Methods: Tet-on- LMP1 HNE2 cells, as a model, were detected with morphological assay, flowcytometry and Caspase 3 assay after 60Co irradiation with LMP1 induced by doxycycline. The apoptosis in the anti-sense survivin transfected cells was tested. Results: The results showed that, with LMP1 expression, the apoptosis rates from morphological assay and flowcytometry were 32.7%±2.1% and 6.3%, which showed that they were all lower than that without LMP1 expression (66.0%±3.0% and 29.6%). When anti-sense of survivin was induced, the apoptosis rates were 59.0%±3.2% and 3.0% respectively, and caspase 3 activity was 3.78 nmol/106 cells, which were higher than that of the control (26.0%±2.6%, 8.6% and 2.79 nmol/106). Survivin restrained the cell apoptosis induced by irradiation, but anti-sense of survivin could release this inhibition of cell apoptosis triggered by LMP1 expression. Conclusion: LMP1 inhibits the irradiation-induced cell apoptosis via triggering survivin expression. Survivin may be targeted in some certain therapy

  2. A Single Amino Acid Change (Asp 53→ Ala53) Converts Survivin from Anti-apoptotic to Pro-apoptotic

    Science.gov (United States)

    Song, Zhiyin; Liu, Shixin; He, He; Hoti, Naser; Wang, Yi; Feng, Shanshan; Wu, Mian

    2004-01-01

    Survivin is a member of the inhibitor of apoptosis protein (IAP) family that has been implicated in both apoptosis inhibition and cell cycle control. Recently, Survivin has attracted growing attention because of its tumor-specific expression and potential applications in tumor therapy. However, its inhibitory mechanism and subcellular localization remain controversial. Here, we report a novel Survivin mutant Surv-D53A, which displays a function opposite to Survivin and a distinctive subcellular distribution compared with its wild-type counterpart. Surv-D53A was shown to induce apoptosis in a p53-independent manner, indicating that tumor suppressor p53 is not involved in its apoptosis pathway. Surv-D53A was shown to markedly sensitize apoptosis induced by TRAIL, doxorubicin, and RIP3. We also demonstrated that similar to wild-type Survivin, Surv-D53A was localized in cytoplasm in interphase and to midbody at telophase. However, it fails to colocalize in chromosomes with Aurora-B in metaphase as wt-Survivin. Surv-D53A mutant is less stable than wt-Survivin and is degraded more rapidly by ubiquitin-proteasome pathway. Additionally, we found that Surv-D53A interacts with wt-Survivin to form heterodimer or with itself to form mutant homodimer, which may account for the loss of its antiapoptotic function. Finally, unlike Survivin*Survivin, neither Surv-D53A*Survivin nor Surv-D53A*Surv-D53A is able to bind to Smac/DIABLO, which may explain the underlying mechanism for its abolishment of antiapoptotic activity of Survivin. PMID:14699067

  3. Inhibition of Survivin and Aurora B Kinase Sensitizes Mesothelioma Cells by Enhancing Mitotic Arrests

    International Nuclear Information System (INIS)

    Purpose: Survivin, a member of the inhibitor of apoptosis gene family, has also been shown to regulate mitosis. It binds Aurora B kinase and the inner centromere protein to form the chromosome passenger complex. Both Aurora B and survivin are overexpressed in many tumors. In this study, we examined whether irradiation affected survivin and Aurora B expression in mesothelioma cells, and how inhibition of these molecules affected radiosensitivity. Methods and Materials: ZM447439 and survivin antisense oligonucleotides were used to inhibit survivin and Aurora B kinase respectively. Western blot was performed to determine the expression of survivin, Aurora B, phosphorylated-histone H3 (Ser 10), and caspase cleavage. Multinucleated cells were counted using flow cytometry, and cell survival after treatment was determined using clonogenic assay. Results: At 3-Gy irradiation an increase was observed in levels of survivin and Aurora B as well as the kinase activity of Aurora B, with an increase in G2/M phase. The radiation-induced upregulation of these molecules was effectively attenuated by antisense oligonucleotides against survivin and a small-molecule inhibitor of Aurora B, ZM447439. Dual inhibition of survivin and Aurora B synergistically radiosensitized mesothelioma cells with a dose enhancement ratio of 2.55. This treatment resulted in increased formation of multinucleated cells after irradiation but did not increase levels of cleaved caspase 3. Conclusion: Inhibition of survivin and Aurora B induces mitotic cell arrest in mesothelioma cells after irradiation. These two proteins may be potential therapeutic targets for the enhancement of radiotherapy in malignant pleural mesothelioma

  4. Survivin is Not Induced by Novel Taxanes

    OpenAIRE

    Sharifi, Nima; Qi, Jun; Bane, Susan; Sharma, Shubhada; Li, Rui; Robey, Robert; William D Figg; Farrar, William L; Kingston, David G I

    2010-01-01

    Taxanes are a critical component of chemotherapy for breast, prostate, lung and other cancers. Initial or acquired tumor resistance to taxanes is therefore one of the most important issues in oncology. Survivin is a prosurvival gene whose expression is a poor prognostic feature. Survivin is induced acutely upon exposure to taxanes and coordinates resistance to taxane-mediated cell death, although the exact mechanism of taxane-mediated survivin induction is not clear. Here, we describe the syn...

  5. 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

  6. Minocycline is cytoprotective in human trabecular meshwork cells and optic nerve head astrocytes by increasing expression of XIAP, survivin, and Bcl-2

    Directory of Open Access Journals (Sweden)

    Marcus Kernt

    2010-06-01

    Full Text Available Marcus Kernt, Aljoscha S Neubauer, Kirsten H Eibl, Armin Wolf, Michael W Ulbig, Anselm Kampik, Cristoph HirneissDepartment of Ophthalmology, Ludwig-Maximilians-University, Munich, GermanyIntroduction: Primary open-angle glaucoma (POAG is one of the leading causes of blindness. Activation of optic nerve head astrocytes (ONHA and loss of trabecular meshwork cells (TMC are pathognomonic for this neurodegenerative disease. Oxidative stress and elevated levels of transforming growth factor beta (TGFβ play an important role in the pathogenesis of POAG. This study investigates the possible antiapoptotic and cytoprotective effects of minocycline on TMC and ONHA under oxidative stress and increased TGFβ levels.Methods: TMC and ONHA were treated with minocycline 1–150 μM. Possible toxic effects and IC50 were evaluated after 48 hours. Cell proliferation and viability were examined in order to assess the protective effects of minocycline on TMC and ONHA. Expression of Bcl-2, XIAP, and survivin, as well as their mRNA expression, were assessed by real time polymerase chain reaction (RT-PCR and Western Blot analysis 48 hours after treatment with minocycline alone and additional incubation with TGFβ-2 or oxidative stress.Results: Minocycline 1–75 μM showed no toxic effects on TMC and ONHA. Under conditions of oxidative stress, both TMC and ONHA showed an increase in viability and an ability to proliferate when treated with minocycline 20–40 μM. RT-PCR and Western blotting yielded an overexpression of Bcl-2, XIAP, and survivin when TMC or ONHA were treated with minocycline 20–40 μM under conditions of oxidative stress and when additionally incubated with TGFβ-2.Conclusion: Minocycline up to 75 μM does not have toxic effects on TMC and ONHA. Treatment with minocycline 20–40 μM led to increased viability and proliferation under oxidative stress and TGFβ-2, as well as overexpression of Bcl-2, XIAP, and survivin. This protective pathway may help

  7. 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.

  8. Expressions of MFN2 and Survivin in different ovarian tissues%探讨MFN2和Survivin在不同卵巢组织中的表达

    Institute of Scientific and Technical Information of China (English)

    白云; 张红梅; 张金艳; 徐小平

    2011-01-01

    Objective: To explore the expressions and significances of MFN2 and Survivin in different ovarian tissues. Methods: Immunohistochemical method was used to detect the expression levels of MFN2 and Survivin in normal ovarian tissue, benign ovarian tumor and malignant ovarian cancer. Results: The positive staining of MFN2 was yellow brown grains which were presented mainly in cytoplasm. The expression rate of MFN2 in malignant ovarian cancer group showed a increasing trend, there was significant difference ( P < 0. 05) . The positive staining of Survivin was yellow or yellow - brown grains which were presented mainly in cell nucleus. The expression rate of Survivin in malignant ovarian cancer group showed a increasing trend, there was significant difference ( P < 0. 05) . Conclusion: The expression levels of MFN2 and Survivin are high in malignant ovarian cancer tissue.%目的:探讨MFN2和Survivin在不同卵巢组织中的表达及意义.方法:用免疫组化法检测MFN2和Survivin在正常卵巢组织、良性卵巢肿瘤、恶性卵巢癌组的表达情况.结果:①MFN2阳性染色主要位于细胞浆,呈棕黄色颗粒,MFN2在恶性卵巢癌组表达率呈上升趋势,差异均有统计学意义(P<0.05).②Survivin阳性染色主要位于细胞核,呈棕黄色颗粒,Survivin在恶性卵巢癌组表达率呈上升趋势,差异均有统计学意义(P<0.05).结论:MFN2和Survivin在恶性卵巢癌组织中表达较高.

  9. 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

  10. 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.

  11. Expression of Livin, Survivin and Caspase-3 in human brain astrocytoma%Livin、Survivin和Caspase-3在星形细胞瘤中的表达

    Institute of Scientific and Technical Information of China (English)

    赵树鹏; 靳彩玲; 赵新利; 周文科

    2012-01-01

    目的 探讨Livin、Survivin和Caspase-3在星形细胞瘤中的表达.方法 采用免疫组织化学法检测50例星形细胞瘤标本及10例正常脑组织中Livin、Survivin和Caspase-3的表达,分析Livin、Survivin的表达与Caspase-3表达的关系.结果 Caspase-3在正常脑组织中的阳性表达率显著高于在星形细胞瘤(P<0.05),且在Ⅰ~Ⅱ级星形细胞瘤中的阳性表达率高于Ⅲ~Ⅳ级(P<0.05).Livin和Survivin在星形细胞瘤中的阳性表达率显著高于正常脑组织(P<0.05),且在Ⅲ~Ⅳ级星形细胞瘤中的阳性表达率明显高于Ⅰ~Ⅱ级(P<0.05).星形细胞瘤中Livin和Survivin 的表达与Caspase-3蛋白的表达均呈负相关(r分别为-0.520和-0.360,P<0.05).结论 Livin、Survivin和Caspase-3的表达可能与星形细胞瘤的发生及发展有关.%Objective To explore the expression of Livin, Survivin and Caspase-3 in human brain astrocytoma. Methods The expressions of Livin, Survivin and Caspase-3 in fifty astrocytomas and ten normal brain tissues were detected with immunohistochemical method,and the relationships of Caspase-3 with Livin,Survivin were analyzed. Results The positive rate of Caspase-3 in normal tissue was significantly higher than that in human brain astrocytoma (P <0. 05) ,it was higher in gradeⅠand II than in grade Ⅲ and Ⅳ(P <0. 05) . The positive rates of Livin and Survivin in human brain astrocytoma were significantly higher than those in normal brain tissue(P <0. 05) ,and they were higher in grade Ⅲ and IV than in grade I andII (P <0. 05) . The expressions of Livin and Survivin were negatively correlated with those of Caspase-3 in human brain astro-cytoma( r = - 0.520,r = - 0.360 ,P < 0.05). Conclusion Livin,Survivin and Caspase-3 may play an important role in incidence and development of human brain astrocytoma.

  12. 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.

  13. 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.

  14. Survivin antisense compound inhibits proliferation and promotes apoptosis in liver cancer cells

    Institute of Scientific and Technical Information of China (English)

    De-Jian Dai; Cai-De Lu; Ri-Yong Lai; Jun-Ming Guo; Hua Meng; Wei-Sheng Chen; Jun Gu

    2005-01-01

    AIM: To evaluate the effects of survivin on cell proliferation and apoptosis in liver cancer.METHODS: MTT assay was used to generate and optimize phosphorothioate antisense oligonucleotides (ODNs)LipofectamineTM2000 (LiP) compound by varying ODNs (μg):LiP (μL) ratios from 1:0.5 to 1:5. Then, liver cancer cells (HepG2) were transfected with the compound. By using RT-PCR and Western blot, the expression levels of survivin mRNA and proteins were detected in HepG2 cells treated with antisense compounds (ODNs:LiP = 1:4), and compared with those treated with sense compounds (1:4) as control.MTT assay was applied to the determination of cell proliferation in HepG2 cells. Active caspase-3 was evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Laser scanning confocal microscopy was performed to detect the subcellular localization of survivin proteins in treated and untreated cells.RESULTS: Antisense compounds (1:4) down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC50 of 250 nmol/L. Its maximum effect was achieved at a concentration of 500 nmol/L, at whichmRNA and protein levels were down-regulated by 80%.The similar results were found in MTT assay. Antisense compound (1:4)-treated cells revealed increased caspase3-like protease activity compared with untreated cells.Untreated cells as control were primarily negative for the presence of active-caspase-3. As shown by transmission electron microscopy, treated cells with antisense compounds (1:4) resulted in morphological changes such as blebbing and loss of microvilli, vacuolization in the cytoplasm,condensation of the cytoplasm and nuclei, and fragmented chromatin. Immunofluorescence analysis confirmed the presence of survivin protein pool inside the cytoplasm in untreated cells. Labeled-FITC immunofluorescence staining of survivin clearly showed that survivin was distributed mainly in a spotted form inside the cytoplasm. Whereas

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

    Institute of Scientific and Technical Information of China (English)

    张娟; 王蓁

    2011-01-01

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

  16. The expression of survivin in breast cancer and its significance%生存素在乳腺癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    张静; 罗丹

    2015-01-01

    目的:探讨乳腺癌中生存素的表达及其意义。方法:检测48例乳腺癌及29例乳腺良性病变中生存素的表达情况。结果:乳腺良性病变中生存素的阳性表达率24.1%;乳腺癌中生存素的阳性表达率72.9%,差异具有统计学意义(P<0.01)。结论:生存素的异常表达对乳腺癌的发生、进展具有一定的推动作用,且可能对乳腺的良恶性病变有提示作用。%Objective:To investigate the expression of survivin in breast cancer and its significance.Methods:The expression of survivin in 48 cases of breast cancer and 29 cases of benign breast lesions were detected.Results:The positive expression of survivin in benign breast lesions was 24.1% ;while in the breast cancer it was 72.9% ,and the difference was statistically significant(P<0.01).Conclusion:Abnormal expression of survivin has a certain role in promoting and the progression of breast carcinoma,therefore,it may be play a suggestive role in benign and malignant lesions of breast.

  17. Effect of small interfering RNA targeting survivin gene on biological behaviour of bladder cancer

    Institute of Scientific and Technical Information of China (English)

    HOU Jian-quan; HE Jun; WANG Xiao-lin; WEN Duan-gai; CHEN Zi-xing

    2006-01-01

    Background Bladder cancer is the most common type of urinary system tumours. It is frequently associated with genetic mutations that deregulate the cell cycle and render these tumours resistant to apoptosis. Survivin, a newly discovered member inhibitor of apoptosis protein (IAP) family in several human cancers, by inducing cell proliferation and inhibiting apoptosis is frequently activated in bladder cancer. We studied the influence of small interfering RNA (siRNA) targeting survivin on the biological behaviour of bladder cancer cells.Methods A double strand survivin target sequence specific siRNA was designed and synthesized. After transfection of bladder cancer cell line T24 by siRNA/liposome complex with increasing concentrations(50-200 nmol/L), the transfectant cells were intratumourally injected at different doses (5 μg or 50μg). The effects were measured in vitro and in vivo.Results The selected siRNA efficiently down-regulated survivin mRNA expression in a dose and time dependent manner. The maximal effect was achieved at the concentration of 100 nmol/L, at which survivin expression level was down-regulated by 75.91%. The inhibition rate of cell growth was 55.29% (P<0.01) and the markedly increased apoptotic rate was 45.70% (P<0.01). In vivo intratumoural injection of 50 μg siRNA-survivin could notably prevent the growth of bladder cancer (P<0.01) in xenografted animals.Conclusion The application of siRNA-survivin could markedly inhibit survivin expression in bladder cancer cell line by inducing apoptosis and inhibiting the growth of the tumour. It may become a new gene therapy tool for bladder cancer.

  18. 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.

  19. Enhancing microRNA transfection to inhibit survivin gene expression and induce apoptosis: could it be mediated by a novel combination of sonoporation and polyethylenimine?

    Institute of Scientific and Technical Information of China (English)

    CHEN Zhi-yi; LIANG Kun; QIURi-xiang; LUO Liang-ping

    2011-01-01

    Apoptosis is a physiologically essential mechanism of cell and plays an important role in reducing the development and progression of tumors.The appealing strategy for cancer therapy is to target the lesions that induce apoptosis in cancer colls.Survivin,the smallest member of the mammalian inhibitors of the apoptosis protein family,is upregulated in various malignancies to protect cells from apoptosis.Survivin knockdown could induco cancer cell apoptosis and inhibit tumor-angiogenesis.Survivin expression would be silenced by microRNA (miRNA)-mediated RNA interference.However,noninvasive and tissue-specific gene delivery techniques remain absent recently and the utilizations of miRNA expression vectors have been limited by inefficient delivery technique,especially in vivo.On the other hand,safe and promising technologies of gene transfection would be valuable in clinical gene therapy.Successful treatment of gene transfer method would lead to a new and readily available approach in the anticancer research.Sonoporation is an alternative technique of gene delivery that uses ultrasound targeted microbubble destruction to create pores in the cell membrane.Based on our previous studies,in this article,we postulated that the transfection of miRNA could be mediated by the combination of sonoporation and polyethylenimine (PEI) which was one of the most effective poly-cationic gene vectors and enhance the endocytosis of plasmids DNA and hypothesized that the gene silencing and apoptosis induction with miRNA targeting human Survivin would be improved by this novel technique.In our opinion,this novel combination of sonoporation and PEI could enhance targeted gene delivery effectively and might be a feasible,novel candidate for gene therapy.

  20. Survivin, a Promising Gene for Targeted Cancer Treatment.

    Science.gov (United States)

    Shamsabadi, Fatemeh T; Eidgahi, Mohammad Reza Akbari; Mehrbod, Parvaneh; Daneshvar, Nasibeh; Allaudin, Zeenathul Nazariah; Yamchi, Ahad; Shahbazi, Majid

    2016-01-01

    Drawbacks of conventional cancer treatments, with lack of specificity and cytotoxicity using current approaches, underlies the necessity for development of a novel approach, gene-directed cancer therapy. This has provided novel technological opportunities in vitro and in vivo. This review focuses on a member of an apoptosis inhibitor family, survivin, as a valuable target. Not only the gene but also its promoter are applicable in this context. This article is based on a literature survey, with especial attention to RNA interference as well as tumor- specific promoter action. The search engine and databases utilized were Science direct, PubMed, MEDLINE and Google. In addition to cell-cycle modulation, apoptosis inhibition, interaction in cell-signaling pathways, cancer-selective expression, survivin also may be considered as specific target through its promoter as a novel treatment for cancer. Our purpose in writing this article was to create awareness in researchers, emphasizing relation of survivin gene expression to potential cancer treatment. The principal result and major conclusion of this manuscript are that survivin structure, biological functions and applications of RNA interference systems as well as tumor-specific promoter activity are of major interest for cancer gene therapy. PMID:27644605

  1. 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.

  2. Expression of Survivin Protein in Endometrial Carcinoma%Survivin蛋白在子宫内膜癌中的表达研究

    Institute of Scientific and Technical Information of China (English)

    王伟; 储洪娟; 蔡仙丽

    2011-01-01

    Objectives: To study the expression of apoptosis related protein Survivin in endometrial carcinoma.Methods :Immunohistochemical S- P method was performed to detect the expression of Survivin protein in 20 cases of normal endometrium, 20 cases of endometrial atypical hyperplasia, 33 cases of endometrial carcinoma and 2 cases UPSC, and examinad the relationship of the expression with clinicopathological variables.Results: In endometrial carcinoma( include UPSC ) and atypical hyperplasia, there was abundant Survivin immunoreactivity in the nucleus and/or cytoplasm of the endometrial carcinoma cells(70.0%, 87.5% and 100.0% ), which was significantly higher than that in normal endometrium (P <0.05 respectively).But there was no significant difference between the expression or rate of Survivin in carcinoma endometrium and atypical hyperplasia endometrium.Moreover, Survivin expression in endometrial carcinoma was correlated with histological grade and surgical - pathological stage, but not with the invasion to myometrium.Conclusions: Survivin may participate in the onset and progression of endometrial carcinoma through inhibiting apoptosis and promoting proliferation.Survivin could be a new biomarkers in the diagnosis, treatment and prognostic evaluation of endometrial carcinoma.%目的:探讨抗凋亡蛋白Survivin在正常子宫内膜、不典型增生性子宫内膜及子宫内膜癌中的表达.探讨Survivin蛋白在子宫内膜癌发生、发展中的作用及其作为判断预后因子的可行性.方法:应用免疫组织化学S-P法,检测20例正常子宫内膜、20例不典型增生性子宫内膜、33例子宫内膜腺癌及2例子宫内膜乳头状浆液性癌(UPSC)中Survivin蛋白的表达,并结合临床病理特点进行分析.结果:Survivin蛋白在不典型增生的子宫内膜、子宫内膜腺癌及UPSC阳性表达率很高,分别为70.0%、87.5%及100%.三者差异无显著性意义(P>0.05),但均高于在正常子宫内膜中的表达率10

  3. Ionizing radiation activates the HIF-lα-Survivin pathway in human lymphoma cells%电离辐射诱导HIF-1α-Survivin通路在人淋巴瘤细胞中的活化

    Institute of Scientific and Technical Information of China (English)

    乔俏; 李光

    2012-01-01

    Objective To determine the levels of radiation-induced expressions of HIF-1α and Survivin in three human lymphoma cell lines with or without HIF-1a specific antagonist or HIF-1a siRNA transfection and to explore the possible mechanism of cell radioresistance.Methods Namalwa,Ramos,Raji cells were irradiated with X-rays with or without the combination of echinomycin (EC),a HIF-1α specific inhibitor,or transfection of HIF-1α siRNA.Then the expression levels of HIF-1α and Survivin proteins and mRNAs were determined by Western blot and real-time PCR,respectively.Results After irradiation,the expression level of HIF-1α protein was up-regulated at 10-20 h followed by up-regulation of survivin protein at 24 h (t =7.53-31.31,P < 0.01 ) in three lymphoma cell lines.Radiation-induced up-expression of survivin protein was significantly reduced by either HIF-1α inhibition or EC treatment (t =7.21-32.81,P <0.01 ).However,radiation failed to induce up-erpression of survivin at both mRNA and protein levels after transfection with HIF-1α siRNA.Conclusion HIF-1α-Survivin pathway is activated in human lymphoma cells after radiation,which might be involved in the cell radioresistance.%目的 研究电离辐射对人非霍奇金淋巴瘤细胞中HIF-1α-Survivin通路活化状态的影响,探讨恶性淋巴瘤放射抗性的机制.方法 采用Western blot方法检测辐射后3种淋巴瘤细胞中HIF-1α和Survivin蛋白的表达水平,观察应用HIF-1α抑制剂Echinomycin及转染反义HIF-1a siRNA对Survivin蛋白及mRNA表达的影响.结果 在人非霍奇金淋巴瘤细胞中存在HIF-1α和Survivin蛋白的表达,X射线照射后10~20 h出现HIF~1α蛋白表达增加,照射后24 h Survivin蛋白表达增加,与对照组比较差异有统计学意义(t =7.53 ~31.31,P<0.01).与单纯照射组比较,采用HIF-1α抑制剂预处理肿瘤细胞后,Survivin蛋白表达下降,且具有药物浓度依赖性(t=7.21 ~32.81,P<0.01).而转染反义HIF-1α siRNA后,照射未诱导产生survivin

  4. 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.

  5. Elevated expressions of Survivin and VEGF proteins are strong independent predictors of survival in squamous carcinoma of larynx%凋亡抑制蛋白Survivin和血管内皮生长因子高表达是喉鳞状细胞癌生存预后的独立预测因子

    Institute of Scientific and Technical Information of China (English)

    Rong Luo; Weijia Kong; Yanjun Wang

    2008-01-01

    Objective: To study the relationship between Survivin and VEGF proteins in a subgroup of patients with squamous carcinoma of larynx. Methods: 108 cases of squamous carcinoma of larynx with clinical data were collected and expressions of Survivin and VEGF in peripheral blood were investigated by enzyme-linked immunosorbent assay (ELISA). Results: Expressions of Survivin and VEGF were significantly associated with T stage, N stage and metastasis of squamous carcinoma of larynx. The patients with Survivin or VEGF over-expressions presented lower survival rate, respectively, as compared to those of low-expression (P < 0.05). The survival rate in squamous carcinoma of larynx patients with Survivin and VEGF dual over-expressions was significantly lower than that of patients with dual low-expression (P < 0.05). Multivariate analysis indicated that both Survivin and VEGF over-expressions in squamous carcinoma of larynx peripheral blood samples were strong independent factors of poor prognosis in squamous carcinoma of larynx patients. Conclusion: Survivin and VEGF over-expressions are independent prognostic factors for the patients with squamous carcinoma of larynx. These results also suggest that peripheral blood Survivin and VEGF expressions are valuable prognostic markers for prognosis prediction in squamous carcinoma of larynx patients.

  6. Relationship between the expression of ERCC-1,survivin and both efficiency and prognosis of cisplatin contained first-line chemotherapy in advanced lung adenocarcinoma%ERCC-1、sruvivin表达与含顺铂方案-线化疗冶疗晚期肺腺癌患者疗效和预后关系

    Institute of Scientific and Technical Information of China (English)

    Xiaobing Chen; Suxia Luo; Ning Li; Yongping Song; Junjie Zen

    2009-01-01

    Immunohistochemical (IHC) method was used to evaluate the expression of ERCC-1 and survivin in 80 pathologically confirmed advanced lung adenocarcinoma patients given cisplatin-contained regimens first-line chemotherapy. The response rate and survival time were analyzed according to the expression of ERCC-1 and survivin. Results:Only 77 patients could be reviewed by IHC staining. The expression rates of ERCC-1 and survivin were 33.77% and 53.25 % respectively. The worse response rate and shorter TTP/PFS could be identified in ERCC-1 positive group. Patients with positive expression of survivin had worse survival time. Conclusion:Expression of ERCC-1 may be a molecular marker of cisplatincontained regimens first-line chemotherapy resistance and poor prognosis in advanced lung adenocarcinoma patients. Positive expression of survivin predicates poor prognosis for patients with advanced lung adenocarcinoma.

  7. Polymeric micelles containing reversibly phospholipid-modified anti-survivin siRNA: a promising strategy to overcome drug resistance in cancer.

    Science.gov (United States)

    Salzano, G; Riehle, R; Navarro, G; Perche, F; De Rosa, G; Torchilin, V P

    2014-02-28

    The discovery that survivin, a small anti-apoptotic protein, is involved in chemoresistance, opens a new scenario to overcome the drug resistance in cancer. It was shown that siRNA can efficiently inhibit the expression of survivin in cancer cells. However, the clinical use of siRNA is still hampered by an unfavorable pharmacokinetic profile. To address this problem, earlier we developed a novel system to deliver siRNA into cancer cells. Namely, we reversibly modified the survivin siRNA with a phosphothioethanol (PE) portion via a reducible disulfide bond and incorporated the resulting siRNA-S-S-PE conjugate into nanosized polyethyelene glycol 2000-phosphatidyl ethanolamine (PEG2000-PE)-based polymeric micelles (PM), obtaining survivin siRNA PM. The activity of these nanopreparations was evaluated by survivin protein down-regulation, tumor cell growth inhibition, and chemosensitization of the treated tumor cells to paclitaxel (PXL). We found a significant decrease of cell viability and down-regulation of survivin protein levels after treatment with survivin siRNA PM in several cancer cell lines. In addition, the down-regulation of survivin by treating cells with survivin siRNA PM, elicited a significant sensitization of the cells to PXL, in both sensitive and resistant cancer cell lines. Finally, we demonstrated successful co-delivery of PXL and survivin siRNA in the same PM leading to superior therapeutic activity compared to their sequential administration. Our results support the use of this new platform for the treatment of the most aggressive tumors. PMID:24099916

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

    Science.gov (United States)

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

    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 wild-survivin 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. PMID:21472129

  9. 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.

  10. 生存素在瘢痕疙瘩组织中的表达及其意义%Expression of survivin in keloid and its clinical significance

    Institute of Scientific and Technical Information of China (English)

    陈明春; 范飞翔; 郭庆; 曾凡钦

    2008-01-01

    Objective To investigate the expression of survivin in keloid, and its significance in the development of keloid. Methods Tissue samples were obtained from 25 patients with keloid (12 males and 13 females, aging from 4 to 44 years with a disease course of 1-18 years), and 15 normal skin samples obtained from surgical operation served as the controls. Streptavidin-biotin-peroxidase complex (SABC) method was applied to detect the expression of survivin in these samples. Results No expressin of survivin was observed in normal controls, while it was expressed in 80.0% (20/25) of the keloid samples with the predominant distribution in fibroblasts and vascular endothelial cells. The positivity rates of survivin were 57.14% (4/7), 81.81% (9/11) and 100% (7/7) in tissues of low-grade, moderate-grade and high-grade keloid, respectively, with no significant difference among the three groups (P = 0.133 ). Similarly, no signifi- cant difference was observed in the positivity rate of survivin between recurrent patients and untreated patients (90.91% (10/11 ) vs 71.43% (10/14), P = 0.341 ). Conclusion Survivin might play a role in the development of keloid.%目的 探讨生存素在瘢痕疙瘩组织中的表达及在瘢痕疙瘩发病中的作用. 方法 采用免疫组化SABC法,检测25例瘢痕疙瘩(病例组)及15例正常皮肤(对照组)组织内生存素的表达情况. 结果 25例瘢痕疙瘩组织中生存素表达阳性20例,阳性率80.0%,表达部位主要在成纤维细胞和血管内皮细胞内,正常皮肤组织内均表达阴性,两组比较差异有统计学意义(X2=24.00,P<0.01).瘢痕疙瘩临床分级低度、中度、重度生存素表达阳性率分别为57.14%(4/7)、81.81%(9/11)、100%(7/7),差异无统计学意义(P=0.133);瘢痕疙瘩复发组与未治疗组生存素表达阳性率分别为90.91%(10/11)、71.43%(10/14),差异亦无统计学意义(P=0.341). 结论 生存素可能参与瘢痕疙瘩的发病.

  11. 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

  12. 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.

  13. Early diagnostic value of survivin and its alternative splice variants in breast cancer

    International Nuclear Information System (INIS)

    The inhibitor of apoptosis (IAP) protein Survivin and its splice variants are differentially expressed in breast cancer tissues. Our previous work showed Survivin is released from tumor cells via small membrane-bound vesicles called exosomes. We, therefore, hypothesize that analysis of serum exosomal Survivin and its splice variants may provide a novel biomarker for early diagnosis of breast cancer. We collected sera from forty breast cancer patients and ten control patients who were disease free for 5 years after treatment. In addition, twenty-three paired breast cancer tumor tissues from those same 40 patients were analyzed for splice variants. Serum levels of Survivin were analyzed using ELISA and exosomes were isolated from this serum using the commercially available ExoQuick kit, with subsequent Western blots and immunohistochemistry performed. Survivin levels were significantly higher in all the breast cancer samples compared to controls (p < 0.05) with exosome amounts significantly higher in cancer patient sera compared to controls (p < 0.01). While Survivin and Survivin-∆Ex3 splice variant expression and localization was identical in serum exosomes, differential expression of Survivin-2B protein existed in the exosomes. Similarly, Survivin and Survivin-∆Ex3 proteins were the predominant forms detected in all of the breast cancer tissues evaluated in this study, whereas a more variable expression of Survivin-2B level was found at different cancer stages. In this study we show for the first time that like Survivin, the Survivin splice variants are also exosomally packaged in the breast cancer patients’ sera, mimicking the survivin splice variant pattern that we also report in breast cancer tissues. Differential expression of exosomal-Survivin, particularly Survivin-2B, may serve as a diagnostic and/or prognostic marker, a “liquid biopsy” if you will, in early breast cancer patients. Furthermore, a more thorough understanding of the role of this

  14. The study on the induction of specific immune cytotoxic T lymphocyte responses against pancreatic cancer by transfected dendritic cells with common tumor antigen survivin mRNAs in vitro%Survivin mRNA转染树突细胞诱导特异性抗胰腺癌免疫反应的体外研究

    Institute of Scientific and Technical Information of China (English)

    郭晓钟; 陈江

    2011-01-01

    Objective To investigate the induction of specific anti-tumor immune response by transfected dendritic cells (DCs) with survivin mRNA of human pancreatic cancer, and to provide the experimental evidences for the treatment of human pancreatic cancer with DCs vaccine. Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs). After being transcripted and amplified, survivin mRNA was transfected into DCs by electroporation. The expression of survivin in DCs at different time points was detected by quantitative real-time PCR. The survival rate of DCs before and after transfection was determined by MTT method. The induction of specific cytotoxic T lymphocyte (CTL) response by survivin mRNA transfected DCs was measured by 51Cr standard cytotoxicity test. The induction of specific CTL activation by survivin mRNA transfected DCs was evaluated through testing released IFN-γ by ELISA method. Results After survivin mRNA transfection for 48h, the expression of survivin mRNA in DCs reached the highest point (46.09±6.57). After transfection, the survival rate of DCs was stabilized around 80%. The DCs transfected with survivin mRNA could effectively induce HLA-A2+ / survivin+ specific CTL immune responses. Stimulated with pancreatic cancer cell line Capan-2 cells or SCL-1 cells as control group, the IFN-γ released in 24 hours by survivin specific CTL were (28.79±5.70) U/ml and (25.12±2.13) U/ml respectively, there was no significant difference (P=0.761). Conclusion The induction of CTLs by DCs transfected with human pancreatic cancer survivin mRNA could produce specific anti-tumor immunity.%目的 研究人胰腺癌survivin mRNA转染树突细胞(DC)诱导的特异性抗肿瘤免疫反应,为DC疫苗治疗胰腺癌提供实验依据.方法 自样本外周血单核细胞中分离和培养DC.体外转录和PCR扩增survivin mRNA后使用电穿孔法将其转染DC.采用实时定量PCR技术检测不同时间点DC中survivin的表达.用四甲基

  15. 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.

  16. Expression of HIF-1a Gene and Survivin Gene in Squamous Cell Carcinoma of Cervix%宫颈鳞癌SiHa细胞中HIF-1a基因、survivin基因的表达

    Institute of Scientific and Technical Information of China (English)

    古力米热·布然江; 艾力克木·阿不都玩克; 李小文

    2015-01-01

    Objective Discussion SiHa cervical squamous carcinoma cel s HIF -1α gene (HIF-1α) relationship with survivin's anti-apoptotic gene survivin gene. Methods This study through with cobalt chloride simulated hypoxia to promote the expression of HIF-1a in cultured cervical squamous cel carcinoma cel line SiHa, again by RNA interference inhibits the expression of HIF-1a gene, by Western blot method was used to detect the HIF-1a gene and survivin gene in the lack of oxygen condition expression. Results After incubation under hypoxic state and increased expression of HIF-1αin cervical squamous cel carcinoma SiHa cel s, survivin protein expression was also increased, and with the lack of oxygen concentration dependent;RNA interference to reduce the expression method of inhibiting HIF-1α gene in cel s HIF-1α , survivin protein expression is reduced. Conclusion In squamous cel carcinoma SiHa cel s, the gene expression of HIF-1α and survivin highly relevant, inhibition of HIF-1αexpression of survivin by inhibition of apoptosis may promote tumor cel s.%目的:探讨宫颈鳞癌SiHa细胞中缺氧诱导因子-1α基因(HIF-1α)与抗凋亡基因生存素survivin基因的关系。方法本研究通过培养宫颈鳞癌SiHa细胞用氯化钴模拟缺氧以促进HIF-1a表达,再以RNA干扰技术抑制HIF-1a基因表达,再通过Westernblot法检测其中HIF-1a基因和survivin基因在乏氧状态下的表达。结果氯化钴培养后宫颈鳞癌SiHa细胞的HIF-1a蛋白表达增加,survivin蛋白表达也增加,并与使用的氯化钴的浓度有关。通过RNA干扰技术抑制HIF-1a基因的干扰后的细胞(SiHa/shRNA)中HIF-1a蛋白表达减少,survivin蛋白表达同时减少。结论在宫颈鳞癌SiHa细胞中, HIF-1α基因和survivin表达具有高度的相关性,并且抑制HIF-1α表达可能通过抑制survivin而促进宫颈鳞癌细胞的凋亡。

  17. 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.

  18. 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。

  19. Expression and clinical significance of Survivin and p53 in breast cancer%Survivin和p53在乳腺癌中的表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    马琳琳; 褚为; 王玉倩; 丛宪玲; 张晓梅

    2014-01-01

    目的 探讨乳腺癌患者血清中Survivin抗体的阳性率及其与乳腺癌患者肿瘤组织中P53表达的相关性;两者与临床病例诊断及预后因素分析的相关性.方法 对92例乳腺癌患者的血清和肿瘤组织标本,分别应用酶联免疫吸附试验(ELISA)检测Survivin抗体的含量,免疫组织化学法检测p53的表达,并将这两种细胞因子与淋巴结转移及临床病理特征进行相关分析.结果 Survivin抗体阳性率为15.00% (14/92),p53阳性率为34.78% (32/92).两者与临床病理特征的相关分析结果表明,Survivin抗体和p53高表达与乳腺癌患者淋巴结转移呈负相关(P<0.05).结论 联合检测Survivin和p53蛋白表达有助于预测淋巴结转移及判断预后.%Objective To investigate the positive rate of Survivin antibody expression in serum from breast cancer patients and its relationship with p53 expression in tumor tissues,and the correlation of Survivin and p53 with clinical diagnosis and prognostic factors.Methods The serum and tumor samples were obtained from 92 cases of breast cancer.Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of Survivin antibody in serum,and immunohistochemistry to detect the p53 expression in tumor tissues.The correlation of Survivin and p53 with both tumor lymph node metastasis and clinicopathologic reports was analyzed.Results Positive rate of Survivin antibody and p53 protein was 15.00% and 34.78% respectively.The high expression of Survivin antibody and p53 was negatively correlated with lymph node metastasis in patients with breast cancer.Conclusion Combined detection of Survivin and p53 expression can be used as an indicator to predict lymph node metastasis and prognosis.

  20. Sexual maturation, serum steroid concentrations, and mRNA expression of IGF-1, luteinizing and progesterone hormone receptors and survivin gene in Japanese quail hens.

    Science.gov (United States)

    Shit, N; Sastry, K V H; Singh, R P; Pandey, N K; Mohan, J

    2014-03-15

    In avian species, sexual maturation represents the evidence of start laying, which is a consequence of the development of ovarian follicles. These follicles are the functional reproductive unit whose maturation and viability critically depends on endocrine, paracrine, and autocrine factors beyond the signals from the central nervous system. The present study was undertaken to investigate the correlation of sexual maturity with tissue growth, mRNA expression of certain genes, and serum steroid concentrations in Japanese quail hens. To carry out the present study, a total of forty Japanese quail hens (5 weeks) were housed individually under uniform husbandry condition with ad libitum quail layer ration and water at 14-hour photo schedule. On sixth week onwards, four birds were sacrificed at each time on 1, 3, 7, 10, 13, 16, 19, 22, 25, and 28 days. Serum was extracted aseptically to analyze the gonadal steroid hormones (estrogen and progesterone) and corticosterone to investigate the liaison with sexual maturation of the species. Expression analyses of four genes i.e., insulin-like growth factor-1, luteinizing hormone receptor, progesterone receptor, and survivin were carried out in the three largest ovarian yellow follicles. A significant (P < 0.05) increase in body weight gain and oviduct weight was recorded during the phase of sexual maturation. Smaller follicles revealed higher insulin-like growth factor-1 and survivin gene expression, whereas the reverse result was manifested in both the luteinizing and progesterone hormone receptors. In biochemical study, the gonadal steroids (estrogen and progesterone) were recorded higher at the first half of the experiment when a gradual decrease in corticosterone concentration was confirmed from the very beginning of this study. This result substantiated that sexual maturation in Japanese quail may be completed by the time of 8 weeks after its birth in support of the analyzed information studied in the current investigation

  1. 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.

  2. 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.

  3. 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.

  4. Multifunctional Polymeric Micelles Co-loaded with Anti-Survivin siRNA and Paclitaxel Overcome Drug Resistance in an Animal Model of Ovarian Cancer.

    Science.gov (United States)

    Salzano, Giuseppina; Navarro, Gemma; Trivedi, Malav S; De Rosa, Giuseppe; Torchilin, Vladimir P

    2015-04-01

    Ovarian cancer is a dreadful disease estimated to be the second most common gynecologic malignancy worldwide. Its current therapy, based on cytoreductive surgery followed by the combination of platinum and taxanes, is frequently complicated by the onset of multidrug resistance (MDR). The discovery that survivin, a small antiapoptotic protein, is involved in chemoresistance provided a new prospect to overcome MDR in cancer, because siRNA could be used to inhibit the expression of survivin in cancer cells. With this in mind, we have developed self-assembly polymeric micelles (PM) able to efficiently co-load an anti-survivin siRNA and a chemotherapeutic agent, such as paclitaxel (PXL; survivin siRNA/PXL PM). Previously, we have successfully demonstrated that the downregulation of survivin by using siRNA-containing PM strongly sensitizes different cancer cells to paclitaxel. Here, we have evaluated the applicability of the developed multifunctional PM in vivo. Changes in survivin expression, therapeutic efficacy, and biologic effects of the nanopreparation were investigated in an animal model of paclitaxel-resistant ovarian cancer. The results obtained in mice xenografed with SKOV3-tr revealed a significant downregulation of survivin expression in tumor tissues together with a potent anticancer activity of survivin siRNA/PXL PM, while the tumors remained unaffected with the same quantity of free paclitaxel. These promising results introduce a novel type of nontoxic and easy-to-obtain nanodevice for the combined therapy of siRNA and anticancer agents in the treatment of chemoresistant tumors. PMID:25657335

  5. 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. PMID:24842412

  6. 小干扰RNA对人肝癌细胞株HepG2Survivin基因表达的影响%Study on siRNA gene transfection on expression of endogenous survivin in hepatoma carcinoma cell line HepG2 cells

    Institute of Scientific and Technical Information of China (English)

    薛净; 李瑜元; 聂玉强; 沙卫红; 张亚历; 周殿元

    2012-01-01

    Objective To study siRNA gene transfection on expression of endogenous survivin in hepatoma carcinoma cell line HepG2 cells. Methods The siRNA gene transfection on expression of endogenous survivin in hepatoma carcinoma cell line HepG2 cells was studied and analyzed. The reverse repeated sequence of survivin siRNA gene was designed and synthesized. Survivin gene sequence specific siRNA vectors were constructed. The hepatoma carcinoma cell line HepG2 cells were transfected with survivin siRNA expression vectors via Lipofectamine TM 2000. MTT assay had been used to detect the rate of cellular inhibition. The expression level of survivin mRNA had been assayed by RT - PCR, morphological observation and flow cytometry analysis. Results There were obvious morphological changes in HepG2 cells after transfection of siRNA under optical microscopy. The cell growth and viability in survivin - siRNA transfected group were significantly inhibited ( P <0.05 ). DNA electrophoresis showed a typical DNA - ladder of apoptosis. The expression of survivin mRNA in cells treated by survivin - siRNA at 24, 48 and 72 h later was significantly reduced by about 56% , 78% and 50% respectively, as compared with those of negative and blank control groups. The latter two groups had similar expression levels. Conclusion Survivin - siRNA designed and transfected in this current study can remarkably inhibit the viability of HepG2 cells and induce their apoptosis, thus it may shed a new experience in gene - therapy for carcinoma.%目的 构建Survivin 基因特异性小干扰RNA(siRNA),检测siRNA-Survivin对Survivin基因表达的抑制,在人肝癌细胞株HepG2中研究 survivin和 survivin siRNA 对细胞凋亡的影响.方法 设计survivin siRNA序列,构建靶向 Survivin siRNA 真核载体.利用脂质体转染人肝癌HepG2细胞,通过相差显微镜下观察、四甲基偶氮唑盐微量酶反应比色法(MTT法)及反转录-聚合酶链反应(RT-PCR)观察

  7. Expression of p73 and its correlation with Survivin in colon carcinoma%结肠癌中P73和Survivin的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    卢笛; 付云辉

    2010-01-01

    Objective To study the expression of P73 and its relationship with expression of Survivin in colon carcinoma. Methods The expression of P73 and Survivin was studied by immunohistochemical technique in 46 cases of colon carcinoma and 12 cases of normal colon mucosa. Results The P73 expression rate of colon carcinoma was significant higher than that of normal colon mucosa(χ2 = 13.42, P <0.01 ). Overexpression of P73 was related to the pathological grade and Dukes stage ( ( χ2 = 8.01, P <0.01 ), and it was closely related to lymphatic metastasis of colon carcinoma( P <0.01 ). There was a positive relation between P73 and Survivin( rs =0.487, P <0.01 ). Conclusions The expression of P73 was closely associated with Survivin in colon carcinoma. The results suggested that the overexpression of P73 and Survivin was involved in the tumorigenesis and development of colon carcinoma and P73 and Survivin might be used as a new tumor marker in metastasis and prognosis of colon carcinoma.%目的 探讨P73和Survivin在结肠癌中的表达及临床意义.方法 应用免疫组化方法检测46例结肠癌组织、12例正常结肠黏膜组织中P73及Survivin的表达.结果 结肠癌与正常结肠黏膜组织中P73阳性表达率分别为67.4%和8.3%,二者阳性率表达比较差异有统计学意义(χ2=13.42,P<0.01);淋巴结转移灶组的P73阳性率明显高于结肠癌原发灶组(P<0.01),P73表达与结肠癌的Dukes分期有关(χ2=8.01,P<0.01).P73与Survivin在结肠癌中的表达呈正相关(rs=0.487,P<0.01).结论 P73与Survivin在结肠癌中的表达密切相关,其参与了结肠癌的发生及发展,可作为判断其转移及预后的参考指标.

  8. 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.

  9. Expressions of PDCD4 and survivin in condyloma acuminatum, Bowenoid papulosis,Bowen disease and squamous cell carcinoma%PDCD4和 survivin 在尖锐湿疣、鲍恩样丘疹病、Bowen 病及鳞状细胞癌皮损中的表达

    Institute of Scientific and Technical Information of China (English)

    于淑萍; 李雪飞; 王丹; 王玉坤

    2015-01-01

    Objective To explore the expressions of programmed cell death 4 (PDCD4)and survivin in patients with condyloma acuminatum (CA),Bowenoid papulosis (BP),Bowen disease (BD)and squamous cell carcinoma (SCC).Methods The expressions of PDCD4 and survivin were detected in normal foreskin (n =10),CA (n =30), BP (n =28),BD (n =10)and SCC (n =11 )with immunohistochemical techniques.Results The expression of PDCD4 in BP,BD and SCC was significantly lower than that in foreskin control (P <0.05),and it was lower in SCC than in CA (P <0.05).The expression of survivin in both BD and SCC was significantly higher than that in control (P <0.05),and it was higher in SCC than in CA (P <0.05).Conclusions PDCD4 can inhibit benign neoplasms from malignant transformation.The overexpression of survivin can inhibit cell apoptosis and promote proliferation.%目的:探讨程序性细胞凋亡因子4(PDCD4)和生存素(survivin)在尖锐湿疣(CA)、鲍恩样丘疹病(BP)、Bowen 病(BD)、鳞状细胞癌(SCC)皮损中的表达及临床意义。方法采用免疫组化 MaxVision 方法检测 CA 组30例、BP 组28例、BD 组10例、SCC 组11例皮损和正常对照组10例中 PDCD4及 survivin 的表达。结果①BP组、BD 组和 SSC 组皮损中 PDCD4表达均较正常对照组低(P <0.05),SCC 组较 CA 组表达低(P <0.05),其余各组间比较差异无统计学意义(P >0.05);②survivin 在 BD 组和 SCC 组的表达高于正常对照组,SCC 组高于 CA组,差异均有统计学意义(P <0.05),其余各组间差异无统计学意义(P >0.05)。结论PDCD4有抑制细胞异常增殖的作用,在皮肤良性增殖性疾病向恶性皮肤肿瘤的转化中发挥一定抑制作用;survivin 过度表达可抑制细胞凋亡、促进细胞增殖。

  10. 免疫组化检测Survivin和Livin表达可预测Dukes' B期结直肠癌术后的复发和生存%The prediction of recurrence and survival of patients with Dukes' B colorectal cancer after curative resection with immunohistochemical detection of Survivin and Livin expressions

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Objective: To detect the expressions of Survivin and Livin in Dukes' B colorectal cancer tissues and analyze the prognosis after curative resection. Methods: The expressions of Survivin and Livin were evaluated immunohistochemically in Dukes' B colorectal cancer specimens from 81 patients after curative resection of the tumor. Their correlations to clinical characters and survival were also explored. Results: The positive rates of Survivin and Livin in colorectal cancer tissues were significantly higher than those in normal colorectal tissues (58.0% vs. 16.7% and 45.7% vs. 8.3% respectively, P <0.05). The expressions of Survivin and Livin were not related to gender, tumor site, primary size, T stage, pathologic category,and degree of differentiation (P > 0.05), and no relationship was found between the expressions of Survivin and Livin (P >0.05). The expression rate of Survivin in patients older than 50 years was higher than that in patients younger than 50 years (70.6% vs. 36.7%, P < 0.05). Both Survivin and Livin were related to recurrence and/or metastasis (P = 0.02 and P = 0.001,respectively), and shorter survival (P = 0.039 and P = 0.001, respectively). Cox multivariate analysis showed T4 and positive Livin expression were independent prognostic factors (P = 0.002 and P = 0.047, respectively). Conclusion: Survivin and Livin are over-expressed in Dukes' B colorectal cancer tissues and are positively related to recurrence and/or metastasis and poor prognosis after curative resection of the tumor.

  11. A Single Amino Acid Change (Asp 53→ Ala53) Converts Survivin from Anti-apoptotic to Pro-apoptotic

    OpenAIRE

    Song, Zhiyin; Liu, Shixin; He, He; Hoti, Naser; Wang, Yi; Feng, Shanshan; Wu, Mian

    2004-01-01

    Survivin is a member of the inhibitor of apoptosis protein (IAP) family that has been implicated in both apoptosis inhibition and cell cycle control. Recently, Survivin has attracted growing attention because of its tumor-specific expression and potential applications in tumor therapy. However, its inhibitory mechanism and subcellular localization remain controversial. Here, we report a novel Survivin mutant Surv-D53A, which displays a function opposite to Survivin and a distinctive subcellul...

  12. 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.

  13. Clinical significance of the expressions of Clusterin, p27 and Survivin in prostate cancer tissues%前列腺癌组织Clusterin与p27及Survivin表达临床意义的研究

    Institute of Scientific and Technical Information of China (English)

    王超奇; 田玉英; 王金花; 梁姗; 梁静; 王璠; 王明; 余大海

    2012-01-01

    目的:探讨凋亡相关蛋白Clusterin、p27和Survivin在前列腺癌组织中的表达及临床意义.方法:应用免疫组织化学方法检测68例前列腺癌组织和47例前列腺增生组织Clusterin、p27和Survivin蛋白的表达.结果:前列腺癌组织Clusterin、p27和Survivin蛋白的阳性表达率分别为75.0% (51/68)、72.1%(49/68)和67.6% (46/68);前列腺增生组织Clusterin、p27和Survivin蛋白的阳性表达率分别为25.5%(12/47)、19.1% (9/47)和23.4% (11/47).Clusterin、p27和Survivin蛋白阳性表达率在前列腺癌组织和前列腺增生组织中差异有统计学意义,P<0.01.Clusterin、p27和Survivin 蛋白表达与前列腺癌病理分级、临床分期和转移情况具有相关性,P<0.05.前列腺癌组织Survivin与Clusterin、Survivin与p27的表达呈正相关,P<0.01.结论:联合检测Clusterin、p27和Survivin蛋白有助于对进展性前列腺癌细胞的侵袭力作出正确评价,以指导临床治疗.%OBJECTIVE : To detect the expressions of Clusterin, p27, Survivin and their correlation In prostate cancer (PCa) tissues. METHODS: Immunohistochemistry was employed to measure the expression of Clusterin,p27 and Survivin in prostatic cancer and 47 cases benign prostatic hyperplasia. RESULTS: In prostatie cancer tissue, the positive expression rates of Clusterin,p27 and Survivin were 75. 0%(51/68) ,72.1%(49/68) and 67. 6%(46/68). In benign prostatic hyper-pksia tissue the positive expression rates of Clusterin, p27 and Survivin were 25. 5% (12/47) ,19, 1% (9/47), 23. 4% (11/47). The positive expression of clusterin,p27 and Survivin between prostatic cancer and benign prostatic hyperplasia were significantly different (P<0.01). The positive expression of Clusterin,p27 and Survivin were related with the histo-logic grade,clinical stage and metastasis of prostatic cancer (P<0. 05). The expression of Surrivin was a positive correlation with clusterin and p27 expression in prostatic cancer (P<0, 01

  14. Aplysin Sensitizes Cancer Cells to TRAIL by Suppressing P38 MAPK/Survivin Pathway

    Directory of Open Access Journals (Sweden)

    Jia Liu

    2014-09-01

    Full Text Available TNF-related apoptosis-inducing ligand (TRAIL is a tumor-selective apoptosis inducer and has been shown to be promising for treating various types of cancers. However, the application of TRAIL is greatly impeded by the resistance of cancer cells to its action. Studies show that overexpression of some critical pro-survival proteins, such as survivin, is responsible for TRAIL resistance. In this study, we found that Aplysin, a brominated compound from marine organisms, was able to restore the sensitivity of cancer cells to TRAIL both in vitro and in vivo. Aplysin was found to enhance the tumor-suppressing capacity of TRAIL on several TRAIL-resistant cancer cell lines. TRAIL-induced apoptosis was also potentiated in A549 and MCF7 cells treated with Aplysin. Survivin downregulation was identified as a mechanism by which Aplysin-mediated TRAIL sensitization of cancer cells. Furthermore, the activation of p38 MAPK was revealed in Aplysin-treated cancer cells, and its inhibitor SB203580 was able to abrogate the promoting effect of Aplysin on the response of cancer cells to TRAIL action, as evidenced by restored survivin expression, elevated cell survival and reduced apoptotic rates. In conclusion, we provided evidence that Aplysin acts as a sensitizer for TRAIL and its effect on p38 MAPK/survivin pathway may partially account for this activity. Considering its low cytotoxicity to normal cells, Aplysin may be a promising agent for cancer treatment in combination with TRAIL.

  15. 大肠癌组织中Survivin和p73的表达及其意义%The expression and significance of Survivin and p73 in human colorectal carcinoma

    Institute of Scientific and Technical Information of China (English)

    赵启军; 王卫

    2011-01-01

    Objective :To investigate the expression and significance of Survivin and p73 in human colorectal caranoma.Methods: S - P immunohistochemical techniquea and image analysis were used to detect the expression of Survivin and p73 in tumor tissue,para - tumor tissue and normal mucos of human colorectal caucinoma.Results : In 60 patients , the average values of light density of Survivin in tumor para - tumor and normal tissus were 0.5443 ±0.0385 ,0.3263 ± 0.0284 ,0.1467 ±0.0227 and p73 were 0.7617 ± 0.0251,0.5327 ± 0.0318 ,0.3460 ± 0.0223respectively.A high expression of Survivin and p73 was discovered in tumor tissue (P <0.01) in tumor tissue,para - tumor tissue and normal mucosa of human colorectal carcinoma.The expression of Survivin and p73 had a linear correlation and the correlation coefficient was 0.783.There was a positive correlation between the expression of Survivin and p73 in human colorectal carcinoma.Conclusion : Survivin and p73 might play an important role in the development of human colorectal carcinoma,and Survivin and p73 had a coordinated effect in the course of development of human colorectal carcinoma.%目的:探讨大肠癌组织中Survivin和p73的表达特点及其相互关系.方法:运用S-P免疫组织化学法染色结合图像分析,探讨Survivin和p73在大肠癌组织、癌旁组织及正常组织中的表达特点.结果:在60例大肠癌的癌、癌旁、正常组织标本中, Survivin表达的平均光密度值分别为 :0.5443±0.0385、0.3263±0.0284、0.1467±0.0227;癌组织中Survivin的表达水平显著升高,与癌旁组织、正常组织中的表达比较存在显著差异(P<0.01).p73表达的平均光密度值分别为:0.7617±0.0251、0.5327±0.0318、0.3460±0.0223;癌组织中p73蛋白的表达水平显著升高,与癌旁组织、正常组织中的表达比较存在显著差异(P<0.01)Survivin和p73在大肠癌组织中的表达存在依从关系(P<0.01),二者呈直线相关,相关系数为0.783.结论

  16. Survivin、 COX-2蛋白在眼睑基底细胞癌中的表达及意义%Expression and Significance of Survivin and COX-2 in Blepharal Basal Cell Carcinoma

    Institute of Scientific and Technical Information of China (English)

    叶青; 杨桂芳

    2011-01-01

    Objective: To research the expression and clinical significance of COX-2 and Survivin protein in eyelid basal cell carcinoma. Methods: The specimens of 100 cases of eyelid basal cell carcinoma and 100 normal specimens from floppy eyelid syndrome patients were collected. Immunohisto-chemical method was adopted to detect the expression of Survivin and COX-2, and the expression results were compared between the two groups. Results: Eyelid basal cell carcinoma showed high COX-2 and Survivin expression, while normal eyelid tissue showed low COX-2 and Survivin expression (both P>0. 05). Conclusion; Survivin and COX-2 are highly expressed and play an important role in eyelid basal cell carcinoma.%目的:探讨COX-2和Suvrivin蛋白在眼睑基底细胞癌组织中的表达及临床意义.方法:搜集本院病理科2000年1月-2010年12月保存的基底细胞癌标本100例,所有眼睑基底细胞癌肿瘤组织均经病理科检查确诊.同时搜集同期100例正常眼睑部组织,正常对照组100例均为单侧,均来自眼睑松弛症患者.运用免疫组织化学方法检测Survivin蛋白和COX-2蛋白在100例眼睑基底细胞癌中的表达.结果:眼睑基底细胞癌组织中COX-2蛋白呈高表达;正常眼睑组织中COX-2蛋白呈低表达.眼睑基底细胞癌组织中Suvrivin蛋白呈高表达;正常眼睑组织中Suvrivin蛋白呈低表达.两种基因表达均无年龄、性别、生长部位的统计学差异(均P>0.05).结论:COX-2、Survivin在眼睑基底细胞癌中的高表达,对眼睑基底细胞癌的发生和发展起了重要作用.

  17. 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

  18. Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    Guo-Ying Miao; Qi-Ming Lu; Xiu-Lan Zhang

    2007-01-01

    AIM: To investigate the inhibitory effect of a specific small survivin interfering RNA (siRNA) on cell proliferation and the expression of survivin in human gastric carcinoma cell line SGC-7901.METHODS: To knockdown survivin expression, a small interfering RNA targeting against survivin was synthesized and transfected into SGC-7901 cells with lipofectamineTM2000. The downregulation of survivin expression at both mRNA and protein levels were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. Cell proliferation inhibition rates were determined by methyl thiazolyl tetrazolium (MTT) assay. The effect of survivin siRNA on cell cycle distribution and cell apoptosis was determined by flow cytometry (FCM).RESULTS: RNA interference could efficiently suppress the survivin expression in SGC-7901 cells. At 48 h after transfection, the expression inhibition rate was 44.52% at mRNA level detected by RT-PCR and 40.17% at protein level by Western blot analysis. Downregulation of survivin resulted in significant inhibition of tumor cell growth in vitro. The cell proliferation inhibition rates at 24, 48 and 72 h after survivin siRNA and non-siliencing siRNA transfection, were 34.06%, 47.61% and 40.36%,respectively. The apoptosis rate was 3.56% and the number of cells was increased in G0/G1 phase from 38.2% to 88.6%, and decreased in S and G2/M phase at 48 h after transfection.CONCLUSION: Downregulation of survivin results in significant inhibition of tumor growth in vitro. The inhibition of survivin expression can induce apoptosis of SGC-7901 cells. The use of survivin siRNA deserves further investigation as a novel approach to cancer therapy.

  19. The correlation of PSA and expression of Clusterin and Survivin in prostate cancer tissue%前列腺癌组织Clusterin与Survivin表达与PSA 的相关性分析

    Institute of Scientific and Technical Information of China (English)

    王超奇; 侯建全

    2015-01-01

    Objective:To study the correlation of PSA and expression of Clusterin and Survivin in prostate cancer tissue. Methods:Immunohistochemistry was employed to measure the expression of Clusterin and Survivin in 80 cases of prostatic cancer and 50 cases of benign prostatic hyperplasia( BPH)and PSA in plasma. Results:In prostatic canc-er tissue,the positive expression rates of Clusterin and Survivin were 57. 5%(46/80)and 48. 8%(39/80). In be-nign prostatic hyperplasia tissue ,the positive expression rates of Clusterin and Survivin were 0( 0/50 ),0( 0/50 ) ( p0. 001). Conclusion:The expression of Clusterin and Survivin and PSA in PCa may offer a new mode for the prevention and treatment of prostatic cancer.%目的:分析蛋白Clusterin和Survivin在前列腺癌( PCa)组织中的表达与前列腺特异性抗原( PSA)的相关性及在前列腺癌诊断中的应用价值。方法:应用免疫组织化学方法检测80例前列腺癌和50例前列腺增生组织凋亡蛋白 Clusterin、Survivin的表达和相应患者血浆中PSA值。结果:前列腺癌组织中Clusterin、Sur-vivin蛋白的阳性表达率分别为57.5%(46/80)、48.8%(39/80),Clusterin和Survivin蛋白在前列腺增生组织中的阳性表达率分别为0(0/50)、0(0/50),差异具有统计学意义( p0.001)。结论:联合检测Clusterin及Survivin表达与前列腺特异抗原( PSA)可为前列腺癌的防治和治疗提供新的思路。

  20. Correlation of the expression of survivin and caspase-3 proteins in juvenile laryngeal papilloma%儿童喉乳头状瘤组织中Survivin及Caspase-3蛋白表达及相关性研究

    Institute of Scientific and Technical Information of China (English)

    汪武; 周远大; 何海霞

    2009-01-01

    目的:研究儿童喉乳头状瘤中survivin及caspase-3蛋白的表达及其相互关系.方法:应用免疫组化方法对儿童喉乳头状瘤组织、儿童声带小结组织及正常的喉黏膜组织中survivin及caspase-3的表达进行检测并进行相关性分析.结果:42例儿童喉乳头状瘤组织中survivin阳性表达率为57.14%, 明显高于儿童声带小结组(P<0.01)和正常黏膜组(P<0.01);42例喉乳头状瘤组织中caspase-3阳性表达率为26.19%, 显著高于儿童声带小结组和正常喉黏膜组 (P<0.01);Spearman相关性分析显示, 儿童喉乳头状瘤中survivin与caspase-3的表达呈显著负相关(r=-0.682, P<0.01).结论:Survivin的高表达和caspase-3的低表达可能在儿童喉乳头状瘤发生发展过程中起着重要作用.%AIM: To investigate correlation between the expression of survivin and caspase-3 proteins in juvenile laryngeal papilloma. METHODS: The expression of survivin and caspase-3 proteins were detected with immunohistochemial method in 43 cases of juvenile laryngeal papilloma, 25 vocal nodules and 25 normal laryngeal mucosa. RESULTS: The positive rates of survivin protein in juvenile laryngeal papilloma were 57.14% and higher than that in voeal nodules (P<0.01)and the normal laryngeal moeusa (P<0.01). And the Caspase-3 protein positive rate was 26.19% in juvenile laryngeal papilloma and lower than that in voeal nodules and the normal laryngeal mucosa (P<0.01).There was a significant negative correlation between the expression of survivin and caspase-3 in juvenile laryngeal papilloma. CONCLUSION: The abnormal expression of survivin and caspase-3 may play important role in the pathogenesis of juvenile laryngeal papilloma.

  1. The effect of amifostine on Survivin expression in HL-60 cells in vitro%亚砷酸联合阿米福汀对HL-60细胞中Survivin表达的影响

    Institute of Scientific and Technical Information of China (English)

    武婕萍; 马梁明; 周永安; 朱镭

    2008-01-01

    目的 探讨亚砷酸(As2O3)联合阿米福汀(AMI)诱导人髓系白血病细胞HL-60凋亡的可能机制.方法 不同浓度As2O3,单用和联合AMI对HL-60细胞进行不同时间干预,用MTT比色法检测细胞的生长抑制作用,用半定量RT-PCR法检测抑凋亡基因Survivin mRNA的表达水平.结果 As2O3组和联合组均可显著抑制HL-60细胞的增生,呈浓度依赖性,联合组对其抑制作用明显大于As2O3组.联合组降低Survivin的表达作用比As2O3组更明显.结论 As2O3通过下调抑凋亡基因Survivin的表达诱导HL-60凋亡;AMI可增强As2O3对Survivin的下调作用,增强HL-60细胞对As2O3的敏感性,发挥促凋亡效应.%Objective To explore the mechanisms of apoptosis induced by arsenic trioxide and amifostine in human acute promyelocytic leukemia cell lines HL-60 in vitro.Methods HL-60 cells were treated with different concentrations of arsenic trioxide alone and combined with amifostine.The inhibitory ratio of the ceils were measured by MTT assay.and the expression of Survivin Was detected by semiquantitate RT-PCR.Results Proliferation of HL-60 cells exposed to arsenic trioxide dwpped down with increasing dose of the dmg and this effect Was significantly hisher when arsenic trioxide Was used in combination with amifostine.Furthermore.there was a more significant decrease in Survivin expression in HL-60 cells treated with arsenic trioxide in combination with amifostine as compared to the cells treated only with arsenic trioxide.Conclusion Arsenic trioxide induced HL-60 cells to undergo apoptosis by downregulating the expression of Survivin. Amifostine enhanced the sensitivity of HL-60 cells to arsenic trioxide by downregulating the expression of Survivin,thus promoting apoptosis effect.

  2. Expression and significance of Survivin, PTEN and mutant type P53 in human brain astrocytom%Survivin和PTEN与P53在人脑星形细胞瘤的表达及意义

    Institute of Scientific and Technical Information of China (English)

    张信芳

    2011-01-01

    Objective To investigate the expressions of apoptosis inhibitor gene-Survivin and tumor suppressor gene-PTEN ,mutant type P53 in human brain astrocytoma and the relationship between these genes and the pathological features of astrocytoma. Methods SP immunohistochemical method was used to detect the expression of Survivin,PTEN and mutant type P53 gene in 15 cases of normal brain tissue and 40 cases of cerebral astrocytoma. Results The positive rates of Survivin,PTEN and mutant type P53 in normal brain tissue were 0%,100% and 0% respectively. The positive rates in astrocytoma were 52.5% ,60.0% and 45.0% respectively. There was a significant difference in the expressions of Survivin,PTEN and mutant type P53 between normal brain tissue and astrocytoma. The expressions of Survivin,P53 were increased with the increase of tumor's grade,however,the expression of PTEN was decreased. Conclusion Survivin, PTEN and mutant type P53 in the occurrence and development of astrocytoma play an important role and the positive rates of Survivin, PTEN and mutant type P53 in astrocytoma are correlated with astrocytoma's grade.%目的 探讨凋亡抑制基因Survivin和抑癌基因PTEN、P53在人脑星形细胞瘤中的表达及与临床病理特征之间的关系.方法 应用SP免疫组织化学法,检测Survivin、PTEN与P53基因在15例正常脑组织(对照组)、40例脑星形细胞瘤(脑星形细胞瘤组)中的表达情况.结果 在正常脑组织和脑星形细胞瘤中 Survivin的阳性率为0%、52.5%;PTEN的阳性率为100%、60.0%;P53的阳性率为0%,45%.2组差异有统计学意义(P<0.05 或<0.01).Survivin、PTEN与P53在脑星形细胞瘤不同分级中有不同的表达,Survivin、P53随着分级的增加表达的阳性率增加,而PTEN随着分级的增加其表达的阳性率降低.结论 Survivin、PTEN与P53在其发生发展过程中起着一定的作用,并与人脑星形细胞瘤的分级相关.

  3. Loss of Survivin in Intestinal Epithelial Progenitor Cells Leads to Mitotic Catastrophe and Breakdown of Gut Immune Homeostasis.

    Science.gov (United States)

    Martini, Eva; Wittkopf, Nadine; Günther, Claudia; Leppkes, Moritz; Okada, Hitoshi; Watson, Alastair J; Podstawa, Eva; Backert, Ingo; Amann, Kerstin; Neurath, Markus F; Becker, Christoph

    2016-02-01

    A tightly regulated balance of proliferation and cell death of intestinal epithelial cells (IECs) is essential for maintenance of gut homeostasis. Survivin is highly expressed during embryogenesis and in several cancer types, but little is known about its role in adult gut tissue. Here, we show that Survivin is specifically expressed in transit-amplifying cells and Lgr5(+) stem cells. Genetic loss of Survivin in IECs resulted in destruction of intestinal integrity, mucosal inflammation, and death of the animals. Survivin deletion was associated with decreased epithelial proliferation due to defective chromosomal segregation. Moreover, Survivin-deficient animals showed induced phosphorylation of p53 and H2AX and increased levels of cell-intrinsic apoptosis in IECs. Consequently, induced deletion of Survivin in Lgr5(+) stem cells led to cell death. In summary, Survivin is a key regulator of gut tissue integrity by regulating epithelial homeostasis in the stem cell niche. PMID:26832409

  4. Loss of Survivin in Intestinal Epithelial Progenitor Cells Leads to Mitotic Catastrophe and Breakdown of Gut Immune Homeostasis

    Directory of Open Access Journals (Sweden)

    Eva Martini

    2016-02-01

    Full Text Available A tightly regulated balance of proliferation and cell death of intestinal epithelial cells (IECs is essential for maintenance of gut homeostasis. Survivin is highly expressed during embryogenesis and in several cancer types, but little is known about its role in adult gut tissue. Here, we show that Survivin is specifically expressed in transit-amplifying cells and Lgr5+ stem cells. Genetic loss of Survivin in IECs resulted in destruction of intestinal integrity, mucosal inflammation, and death of the animals. Survivin deletion was associated with decreased epithelial proliferation due to defective chromosomal segregation. Moreover, Survivin-deficient animals showed induced phosphorylation of p53 and H2AX and increased levels of cell-intrinsic apoptosis in IECs. Consequently, induced deletion of Survivin in Lgr5+ stem cells led to cell death. In summary, Survivin is a key regulator of gut tissue integrity by regulating epithelial homeostasis in the stem cell niche.

  5. 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.

  6. 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

  7. 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.

  8. 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

  9. 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.

  10. 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.

  11. Survivin-specific small interfering RNAs enhance sensitivity of glioma U-87MG cells to paclitaxel by promoting apoptosis

    Institute of Scientific and Technical Information of China (English)

    Yunliang Xie; Yanbo Liu; Weigao Shen; Bo Zhang; Qun Liu

    2012-01-01

    A survivin siRNA expression vector was transfected into glioma U-87MG cells and these cells were then treated with paclitaxel. The results showed that survivin-specific siRNA combined with paclitaxel treatment synergistically inhibited glioma U-87MG cell proliferation and promoted apoptosis. This treatment also inhibited the expression of the cell cycle regulatory proteins, survivin, cyclinD1, c-Myc and CDK4 and enhanced the sensitivity of U-87MG cells to paclitaxel.

  12. 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.

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

    International Nuclear Information System (INIS)

    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. 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. 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-independent prostate cancer cell lines. Survivin

  14. Survivin蛋白、CDX2在结直肠癌及癌前病变中的表达和意义%Expression and significance of Survivin protein, CDX2 in colorectal cancer and precancerous diseases

    Institute of Scientific and Technical Information of China (English)

    姚凡保; 付汐; 邝倩仪; 马海生

    2015-01-01

    目的:探讨Survivin蛋白、CDX2在结直肠癌及癌前病变中的表达,为结直肠癌的早期诊断、基因靶点治疗、评估预后、判断复发提供新的标记物。方法收集结直肠癌、腺瘤伴高级别上皮内瘤变、腺瘤伴低级别上皮内瘤变及正常对照组各30例,应用免疫组化方法分别检测各组中Survivin蛋白、CDX2的表达情况。结果 Survivin蛋白表达在腺癌组、高级别上皮内瘤变组、低级别上皮内瘤变组及正常组各组间均有明显差异(P0.05)。Survivin与CDX2的表达呈高度密切负相关(r越-0.64,P0.05). The expression of Survivin was highly negative correlated with CDX2 (r=-0.64, P<0.01). Conclusion Survivin protein and CDX2 are the important biological markers in the develop-ment and progression of colorectal carcinoma, there is a close relationship between them, joint detection of sur-vivin and CDX2 could be a new method for colorectal cancer early diagnosis, gene targeting therapy, evalua-tion prognosisand recurrence.

  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. The study of RNAi-mediated by conditionally replicating adenovirus silencing on Survivin gene in colon cancer cell lastingly

    Institute of Scientific and Technical Information of China (English)

    Chunyi Wang; Zhongxue Fu

    2008-01-01

    Objective: To investigate the effect of RNAi-mediated Survivin gene with conditionally replicating adenovirus silencing on Survivin gene in colon carcinoma cell lines HT-29 lastingly.Methods: We transfected Ad-delElb55KD-shRNA/ Survivin-EGFP to HT-29 (control was replication defective adenovirus and liposome vector which was contained the same shRNA as Ad-delE1b55KD-shRNA/Survivin-EGFP).The expressions of EGFP, Survivin mRNA and Survivin protein in HT29 were detected at the 1st, 7th, 14th and 28th days after transfection.Results: The expression of EGFP, the inhibition of Survivin mRNA and Survivin protein in HT-29 were high in each group at the 7th day after transfection, among the total, the effect of Ad-delE1b55KD-shRNA/Survivin-EGFP group was the highest; at the 14th day, the effects of replication defective adenovirus group and liposome vector group were decreased obviously, and it was still high in Ad-delE1b55KD-shRNA/Survivin-EGFP group; at the 28th day, the effects of control groups were disappeared, and it was still high in Ad-delE1b55KDshRNA / Survivin-EGFP group like before (P<0.05).Conclusion: RNAi-mediated Survivin gene with conditionally replicating adenovirus can silence Survivin gene in colon carcinoma call lines HT-29 lastingly.

  17. 双胰腺癌相关抗原RNA转染树突细胞诱导特异性抗癌免疫反应的实验研究%Induction of specific immune cytotoxic T lymphocyte responses against pancreatic cancer by transfected dendritic cells with MUC4 and Survivin mRNA in vitro

    Institute of Scientific and Technical Information of China (English)

    陈江; 郭晓钟; 李宏宇; 邵晓东; 许文达

    2014-01-01

    Objective To investigate the induction of specific anti-tumor immune response induced by MUC4 and Survivin mRNA co-transfected dendritic cells (DCs) to provide the experimental evidences for the treatment of human pancreatic cancer with multi-epitope loaded DC vaccine. Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs). After being transcripted and amplified, MUC4 and Survivin mRNA were co-transfected into DCs by electroporation. The expression of MUC4 and Survivin in DCs were detected by Western blot. The survival rate of transfected DCs were determined by MTT method. The induction of specific CTL activation by MUC4 and Survivin mRNA co-transfected DCs were evaluated through testing released IFN-γ by ELISA method. The induction of specific cytotoxic T lymphocyte (CTL) re-sponse by MUC4 and Survivin mRNA co-transfected DCs were measured by 51Cr standard cytotoxicity test. Results After MUC4 and Survivin mRNA co-transfection for 72 hours, the expression amount of MUC4 and Survivin were lower than the expression amount of MUC4 or Survivin individually. Compared with the MUC4 or Survivin mRNA individual transfected DCs, the IFN-γreleased in 24 hours by CTLs stimulated with MUC4 and Survivin mRNA co-transfection DCs were (33.84 ± 3.51)U/ml which was significantly higher than the amount of (21.87 ± 4.12)U/ml by MUC4 or (16.61 ± 2.09)U/ml by Survivin mRNA individually (P < 0.05). DCs co-transfection with MUC4 and Survivin mRNA could effectively induce HLA-A2 +/MUC4+/Survivin + specific CTL immune responses against pancreatic cancer cells in vitro. Conclusion The induction of CTLs by DCs co-transfected with human pancreatic cancer MUC4 and Survivin mRNA could produce more powerful specific anti-tumor immunity than single antigen loaded DCs.%目的:研究人胰腺癌MUC4与Survivin mRNA联合转染树突细胞(DC)诱导的特异性抗肿瘤免疫反应,为构建负载多抗原表位DC疫苗治疗胰腺癌提供实验依据

  18. Sequestering survivin to functionalized nanoparticles: a strategy to enhance apoptosis in cancer cells.

    Science.gov (United States)

    Jenkins, Ragini; Bandera, Yuriy P; Daniele, Michael A; Ledford, LeAnna L; Tietje, Ashlee; Kelso, Andrew A; Sehorn, Michael G; Wei, Yanzhang; Chakrabarti, Mrinmay; Ray, Swapan K; Foulger, Stephen H

    2016-04-01

    Survivin belongs to the family of inhibitor of apoptosis proteins (IAP) and is present in most cancers while being below detection limits in most terminally differentiated adult tissues, making it an attractive protein to target for diagnostic and, potentially, therapeutic roles. Sub-100 nm poly(propargyl acrylate) (PA) particles were surface modified through the copper-catalyzed azide/alkyne cycloaddition of an azide-terminated survivin ligand derivative (azTM) originally proposed by Abbott Laboratories and speculated to bind directly to survivin (protein) at its dimer interface. Using affinity pull-down studies, it was determined that the PA/azTM nanoparticles selectively bind survivin and the particles can enhance apoptotic cell death in glioblastoma cell lines and other survivin over-expressing cell lines such as A549 and MCF7 relative to cells incubated with the original Abbott-derived small molecule inhibitor. PMID:26845086

  19. 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

  20. Opposed arsenite-mediated regulation of p53-survivin is involved in neoplastic transformation, DNA damage, or apoptosis in human keratinocytes

    International Nuclear Information System (INIS)

    Highlights: ► Different concentrations of arsenite cause biphasic effects in HaCaT cells. ► p53-survivin signal pathway plays a role in arsenite-induced biphasic effects. ► ERKs inactivate p53, but improve survivin expression by NF-κB/mot-2. ► JNKs block survivin expression by preventing p53 from mdm2-mediated degradation. ► ERKs and JNKs play roles in arsenite-induced biphasic effects. -- Abstract: Biphasic dose–response relationship induced by environmental agents is often characterized with the effect of low-dose stimulation and high dose inhibition. Some studies showed that arsenite may induce cell proliferation and apoptosis via biphasic dose–response relationship in human cells; however, mechanisms underlying this phenomenon are not well understood. Our present study shows that, for human keratinocytes (HaCaT) cells, a low concentration of arsenite activates extracellular signal-regulated kinases (ERKs), which leads to up-regulation of nuclear factor κB (NF-κB) binding to DNA and to elevated, NF-κB-dependent expression of mot-2 (a p53 inhibitor) and survivin (an inhibitor of apoptosis). Activation of p53 is blocked, and neoplastic transformation is enhanced. Inhibition of ERKs reduces cell proliferation and neoplastic transformation. In contrast, a high concentration of arsenite activates c-Jun N-terminal kinases (JNKs), positive regulators of p53, by binding to p53 and preventing its murine double minute 2 (mdm2)-mediated degradation. The elevated levels of p53 lead to repair of DNA damage and apoptosis. Inhibition of JNKs increases DNA damage but decreases apoptosis. By identifying a mechanism whereby ERKs and JNKs-mediated regulation of the p53-survivin signal pathway is involved in the biphasic effects of arsenite on human keratinocytes, our data expand understanding of arsenite-induced cell proliferation, neoplastic transformation, DNA damage, and apoptosis.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. Application of quantum dots as vectors in targeted survivin gene siRNA delivery

    Directory of Open Access Journals (Sweden)

    Zhao JJ

    2013-04-01

    Full Text Available Jianjiang Zhao, Xiaoling Qiu, Zhiping Wang, Jie Pan, Jun Chen, Jiusong Han Department of Surgery, Guangdong Provincial Stomatological Hospital, Southern Medical University, Guangzhou, People's Republic of China Abstract: Gene silencing using short interfering RNA (siRNA is becoming an attractive approach for probing gene function in mammalian cells. This study evaluated the specificity and efficiency of quantum dots (QDs as non-viral gene vectors for delivery of survivin siRNA and downregulation of survivin gene expression in oral squamous cell carcinoma Tca8113 cells. Water-dispersible cationically-modified QDs were electrostatically attached to anionic siRNA molecules and complexed with siRNA for downregulating expression of the survivin gene. Cellular uptake and allocation of QD–siRNA complexes in Tca8113 cells were monitored using confocal laser scanning microscopy. Real-time polymerase chain reaction (PCR was used to quantify survivin messenger RNA (mRNA levels. CdSe QDs were observed with high intensity fluorescence under confocal laser scanning microscopy. Tca8113 cells were successfully transfected by QDs with survivin siRNA, and the red fluorescence from CdSe QDs and green fluorescein amidite fluorescence from siRNA could both be easily observed after 6 hours of incubation. The release of siRNA into the cytoplasm was verified through real-time PCR quantification that showed reduced survivin mRNA levels. In this study, survivin siRNA successfully complexed with water-soluble CdSe QDs and exhibited excellent fluorescent properties and downregulated the expression of the survivin gene in oral squamous cell carcinoma Tca8113 cells. QDs are a novel non-viral gene delivery vector. Keywords: quantum dots, survivin, siRNA delivery, transfection, Tca8113, tongue cancer cells

  6. RNA干扰沉默宫颈鳞癌细胞中HIF-1a基因对survivin基因表达的影响%Effect of RNA Interference Silencing HIF-1a Gene in Cervical Squamous Cell on Carcinoma Cells Survivin Gene Expression

    Institute of Scientific and Technical Information of China (English)

    古力米热·布然江; 艾力克木·阿不都玩克; 李小文

    2015-01-01

    目的:研究 RNA 干扰对宫颈鳞癌 SiHa 细胞中 HIF-1α及survivin 的抑制效应。方法使用不同缺氧时间分别缺氧 SiHa 细胞,以Westernblot 法检测细胞中 HIF-1a 蛋白的表达变化,选择可以引起HIF-1a 蛋白表达增加的缺氧培养时间。进行缺氧后用 Westernblot 法和逆转录聚合酶链反应检测 HIF-1α及 survivin 的表达。结果 SiHa细胞 HIF-1α的表达在缺氧处理时较常氧培养时增加,在缺氧后24~48小时内明显升高,60小时略有降低,72~96 h 逐渐减少。氯化钴培养后 SiHa 细胞的 HIF-1a 蛋白及 survivin 蛋白表达增加,并与氯化钴浓度有关。通过 RNA 干扰技术抑制 HIF-1a 基因的干扰后的细胞中 HIF-1a 及 survivin 表达同时减少。结论在 SiHa 细胞中,HIF-1α基因和 survivin 表达具有高度的相关性,并且抑制 HIF-1α表达可能通过抑制 survivin 而促进癌细胞的凋亡。%Objective To explore the inhibition effect of RNA interference to HIF-1 alpha and survivin in cervical squamous carcinoma SiHa cells. Methods Deprivate oxygen to SiHa cells in different time, then detecte the changing of HIF-1a protein by Westernblot and choosing the right time which can increase the expression of HIF-1a. Use Westernblot and RT-PCR to detect the expression of HIF - 1 alpha and survivin. Results The HIF-1a expression in SiHa cells was increased by oxygen deprivation, and rised in 24~48 h, decreased a little in 60 h and much more in 72~96 h. The expression of HIF-1a and survivin is related to the concentration of CoCl2, and was decreased by using RNA interference. Conclusion In SiHa cells, the expression of HIF-1a is highly related to survivin so that suppression of HIF-1a could promote apoptosis of cancer cells.

  7. 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.

  8. Localization and upregulation of survivin in cancer health disparities: a clinical perspective.

    Science.gov (United States)

    Khan, Salma; Ferguson Bennit, Heather; Asuncion Valenzuela, Malyn May; Turay, David; Diaz Osterman, Carlos J; Moyron, Ron B; Esebanmen, Grace E; Ashok, Arjun; Wall, Nathan R

    2015-01-01

    Survivin is one of the most important members of the inhibitors of apoptosis protein family, as it is expressed in most human cancers but is absent in normal, differentiated tissues. Lending to its importance, survivin has proven associations with apoptosis and cell cycle control, and has more recently been shown to modulate the tumor microenvironment and immune evasion as a result of its extracellular localization. Upregulation of survivin has been found in many cancers including breast, prostate, pancreatic, and hematological malignancies, and it may prove to be associated with the advanced presentation, poorer prognosis, and lower survival rates observed in ethnically diverse populations. PMID:26185415

  9. Inhibition of lymphatic metastases by a survivin dominant-negative mutant.

    Science.gov (United States)

    Xu, Guang-Chao; Zhang, Peng; Leng, Fei; Pan, Li; Li, Zhi-Yong; Yu, Dan-Dan; Shan, Yan; Yuan, Qing-Zhong; Wen, Yuan; Mu, Bo; Shi, Hua-Shan; Chen, Xiang; Wang, Chun-Ting

    2012-01-01

    Metastasis is the most lethal attribute of human malignancy. High-level expression of survivin is involved in both carcinogenesis and angiogenesis in cancer. Previous studies indicate that a mutation of the threonine residue at position 34 (Thr34Ala) of survivin generates a dominant-negative mutant that induces apoptosis, inhibits angiogenesis, and suppresses highly metastatic breast carcinoma in mouse models. We investigated the efficacy of gene therapy with a survivin dominant-negative mutant and possible factors related to lymph node metastasis. The metastasis rate was compared between each group in order to find a survivin-targeted therapy against lymphangiogenesis in its earliest stages. We established lymph node metastasis models and treated animals with H22 tumors with Lip-mSurvivinT34A (Lip-mS), Lip-plasmid (Lip-P), or normal saline (NS). Eight days after the last dose, five randomly chosen mice from each group were sacrificed. We detected the apoptotic index, microvessel density (MVD), lymphatic microvessel density (LMVD), and the expression of VEGF-D with immunohistochemistry. After the remaining animals were sacrificed, we compared the tumor-infiltrated lymph nodes in each group. Administration of mSurvivinT34A plasmid complexed with cationic liposome (DOTAP/chol) resulted in the efficacious inhibition of tumor growth and lymph node metastasis within the mouse H22 tumor model. These responses were associated with tumor cell apoptosis, and angiogenesis and lymphangiogenesis inhibition. Our results suggested that Lip-mSurvivinT34A induced apoptosis and inhibited tumor angiogenesis and lymphangiogenesis, thus suppressing tumor growth and lymphatic metastasis. The mSurvivinT34A survivin mutant is a promising strategy of gene therapy to inhibit lymphatic metastasis. PMID:24139416

  10. 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

  11. 新的细胞凋亡抑制基因survivin在垂体腺瘤中的表达及其与caspase-3表达相关性的研究%Apoptosis- regulating protein,survivin:expression and relationship with caspase- 3 protein in pituitary adenomas

    Institute of Scientific and Technical Information of China (English)

    马杰; 石智勇; 魏冰; 乔思杰

    2002-01-01

    Objective To study the expression of a inhibitor gene,survivin,in pituitary adenomas and its relationship with the expression of caspase- 3 gene.Method Using streptavidin- biotin peroxidase(SP),the expression of surviving,caspase- 3 protein was examined in 8 normal pituitary tissue and 38 pituitary adenomas.Result Survivin was expressed in 23 of 38 cases of pituitary adenomas,in contrast,normal pituitary tissue did not express surviving.Positive expression rate of PRL group,GH group,Mixed group in pituitary adenomas was 12/17,7/13,4/8,respectively.Overexpression of surviving was not related with the tumor type(P >0.05).Meanwhile,surviving positive rate in caspase- 3 positive and negative groups was 5/14,79% .The result showed it was strongly associated with caspase- 3 expression(P<0.05).Conclusions Apoptosis suppression by surviving play an important role in apoptosis of pituitary adenomas.Survivin could be a new diagnostic/therapeutic target in pituitary adenomas.

  12. 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.

  13. 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.

  14. Baseline levels of CD8+ T cells against survivin and survivin-2B in the blood of lung cancer patients and cancer-free individuals.

    Science.gov (United States)

    Karanikas, Vaios; Soukou, Faye; Kalala, Fani; Kerenidi, Theodora; Grammoustianou, Evangelia S; Gourgoulianis, Konstantinos I; Germenis, Anastasios E

    2008-11-01

    Survivin and its variant survivin-2B have been considered as potential candidates for cancer immunotherapy. The magnitude however of spontaneously occurring CD8(+) T cells circulating precursor CTLs (pCTL), has never been evaluated. We set out to measure in 20 patients with lung carcinomas and 5 aged matched healthy male individuals (expressing HLA-A2 and/or -A24), the frequency of pCTLs specific for two naturally processed and presented peptides of survivin (LTLGEFLKL presented by HLA-A2) and survivin-2B (AYACNTSTL presented by HLA-A24) since these peptides are the only ones used in immunotherapeutic trials. The frequency of peptide-specific pCTLs was estimated using a sensitive method that combines HLA-multimer flow cytometric technology with a previous step of in vitro amplification under limiting dilution conditions. Anti-survivin or anti-survivin-2B specific CTL clones were not detected in 17 out of the 21 tested patients, and in none of the healthy individuals. In a number of peripheral blood mononuclear cell microcultures of the remaining 4 patients, diffuse clusters stained weakly by the HLA-multimers were observed which were not amplified after further stimulation and, therefore, they were finally considered as negative. The significance of the levels of spontaneously occurring CTL-responses against survivin and survivin-2B peptides, in cancer patients and cancer-free subjects, remains to be elucidated and it would be interesting to be considered in relation to the clinical efficacy of anti-cancer vaccination protocols. PMID:18789878

  15. 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

  16. 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.

  17. Effect of RNAi targeting survivin gene combined with X-rays radiation on apoptosis of lung adenocarcinoma A549 cells

    International Nuclear Information System (INIS)

    Objective: To construct the vector of RNA interference (RNAi) targeting survivin gene and observe its effect combined with X-rays radiation on lung adenocarcinoma A549 cell apoptosis. Methods: One pair of RNAi sequence targeting survivin gene were designed according to its cDNA sequence reported in GenBank, the recombinant RNAi plasmid pGenesil2-survivin was constructed. After identified by enzyme digestion and sequencing, the pGenesil2-survivin plasmid was trasfeced into A549 cells.In the experiment, normal group,pGenesil2 group, pGenesil2-survivin group,5 Gy irradiation group and pGenesil2-survivin + 5 Gy irradiation group were set up.The apoptosis of A549 cells was measured by flow cytometry with PI/Annexin V and TUNEL,the survivin and caspase-3 expressions were measured by Western blotting. Results: Two fragments about 389 bp and 4 206 bp were gotten by Kpn I and EcoR I enzyme digestion, they are the same to expected result, the sequencing result was compared to oligonucleotide chain with DNAssist 2.0, they were equal, these indicated the identification of pGenesil2-survivin vector was right; pGenesil2-survivin was transfected into A549 cells for 48 h, the apoptotic percentage in pGenesil2-survivin and 5 Gy X-rays groups increased obviously (P< 0.05), when the both were combined, the effect was more obvious;the Western blotting results appeared that the survivin gray scale/β-actin gray scale in pGenesil2-survivin group was lower than that in normal group(P< 0.01), and the caspase-3 gray scale/β-actin gray scale was higher than that in normal group,and that ratio in pGenesil2-survivin+5 Gy irradiation group was more high(P< 0.01). Conclusion: RNAi targeting surviving gene could inhibit survivin protein expression,but enhance caspase-3 protein expression, and promote apoptosis. When it is combined with 5 Gy X-rays irradiation, the promotion of apoptosis is enhanced. (authors)

  18. 直肠癌组织中hPTTG1和survivin的表达及意义%The expression and significance of hPTTG1 and survivin in rectal carcinoma tissue

    Institute of Scientific and Technical Information of China (English)

    殷江霞; 孙红芹; 唐磊; 赵长林; 徐惠绵

    2011-01-01

    Objective: To detect the expression of hPTTCJ1 ( human pituitary - tumour transforming gene, hPTTG1 ) and survivin in rectal carcinoma, rectal adenoma and para - cancerous tissue, explore their correlation and mechanism with biological behavior of rectal cancer. Methods : Sixty specimen of rectal carcinoma were collected from January 2006 to Decemher 2008 in our hospital, immunohistochemical SP staining method was used to detect the expression of hPTTG1 and survivin in 60 cases of rectal carcinoma, 10 cases of rectal adenoma and 10 cases of para cancerous tissues, and analyzed the relationship and hiological behavior among the speciments. Results : The positive rates of hPTTG1 in rectal carcinoma, rectal adenoma and para - cancerous tissues were 80% ( 48/60 ), 20% ( 2/ 10 ) and 0 ( 0/10 ) , respectively , ( P < 0. 01 ). The positive rates of survivin in rectal carcinoma , rectal adenoma and para - cancerous tissue were 90% ( 54/60 ), 80% ( 8/10 ) and 20% ( 2/10 ), respectively ( P < 0. 05 ). The expression of hPTTG1 in rectal carcmoma was significantly correlated with Dukes staging, the degree of differentiation, lymph node metastasis and type of the tumor ( P < 0. 05 ). The expression of survivin was significantly correlated with Dukes staging and lyruph node metastasis in rectal carcinoma ( P < 0. 05 ). The expression of hPTTG1 and survivn in rectal adenocarcmoma was not correlated with gender and age of patients, CEA, tumor locations of rectum. Conclusion: hPTTG1 and survivn may play an important role in development and progression of rectal carcinoma, show a close correlation with rectal carcinoma invasion and metastasis.%目的:检测人垂体瘤转化基因1(human pituitary-tumour transforming gene,hPTTG1)与存活素(survivin)在直肠癌、直肠腺瘤和癌旁正常组织中的表达,分析两者与直肠癌生物学行为的关系和机制.方法:收集2006-2008年间我院直肠癌手术标本60例,直肠腺瘤10例,癌旁正常组织10

  19. Anti-survivin antibody responses in lung cancer.

    Science.gov (United States)

    Karanikas, Vaios; Khalil, Sanaa; Kerenidi, Theodora; Gourgoulianis, Konstantinos I; Germenis, Anastasios E

    2009-09-18

    Existing evidence regarding spontaneous anti-survivin humoral responses in lung cancer is inconclusive. Moreover, despite that cancer cell death elicited by radiotherapy and some chemotherapeutic agents seems to be immunogenic, information about the possible effect of treatment on these responses, is lacking. Serum samples from 33 small cell lung cancer (SCLC) and 117 non-small cell lung cancer (NSCLC) patients upon diagnosis, and from 100 controls, were tested by ELISA for anti-survivin antibodies. Cutoff was set to the mean+2SD of controls. 7.7% of NSCLC, none of the SCLC patients and 2% of the controls appeared with elevated antibody levels (OR 3.6, 95% CI 0.7-17.3 for NSCLC, OR 0.6, 95% CI 0.03-12.6 for SCLC). Measurement of antibodies in 76 NSCLC patients post therapies and during their follow-up, revealed that in 12 NSCLC patients the antibody levels increased up to 2-38 times, and in seven others, they decreased by 2-8 times. No significant correlation was uncovered between either the antibody levels upon diagnosis or their changes post therapies and during follow-up, and any clinicopathological parameter, their response to therapy and survival. Survivin does not induce considerable humoral responses in lung cancer. Potentially, however, strong anti-survivin antibody responses can be elicited during the post therapy and follow-up of the patients, whose clinical significance remains to be elucidated. These findings, together with our previous data concerning survivin expression and the related cytolytic T cell responses in lung cancer, signify a high tolerogenic potential of this tumor-associated antigen. PMID:19380192

  20. 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

  1. Survivin基因在牛蒡子苷元增强人肺癌H460细胞化疗敏感性中的作用%Biological functions of Survivin in chemosensitization of arctigenin in lung cancer H460 cells

    Institute of Scientific and Technical Information of China (English)

    王焕勤; 王静

    2015-01-01

    Objective To explore the role of survivin in the chemosensitization of arctigenin in lung cancer H460 cells.Methods Human full-length Survivin cDNA was amplified by polymerase chain reaction and cloned into pcDNA3.1 (+) expression vector.The resultant survivin-expressing plasmid or empty vector was individually transfected into H460 cells using Lipofectamine 2000.Twenty-four hours after transfection,survivin expression in H460 cells was tested by Western blot.Transfected cells were exposed to arctigenin alone or combined with cisplatin and cell apoptosis was assessed using Annexin-Ⅴ/ PI staining methods.Results Compared to untransfected and empty vector-transfected H460 cells,survivin-transfected cells had a 10-fold elevation in the survivin expression.Low-dose cisplatin did not significantly affect the expression of survivin in H460 cells.In contrast,arctigenin significantly inhibited the expression of survivin protein.When cisplatin was combined with arctigenin,a greater inhibition of survivin expression was observed.Compared to transfection of empty vector,pre-transfection with pcDNA3.1-Survivin plasmid significantly reversed the pro-apoptotic activity of arctigenin alone or combined with cisplatin,leading to a significant reduction in the apoptotic index.Conclusions Arctigenin can inhibit the expression of survivin in lung cancer H460 cells.Arctigenin-mediated chemosensitization of lung cancer H460 cells to cisplatin is associated with suppression of survivin signaling pathway.%目的 研究Survivin基因在牛蒡子苷元促进肺癌细胞化疗敏感性中的作用.方法 应用RT-PCR技术从H460细胞中扩增全长人Survivin cDNA,并将其克隆至pcDNA3.1(+)表达载体,构建人Survivin基因真核表达质粒.将pcDNA3.1-Survivin重组质粒或空载体分别转染H460细胞,孵育24 h后,应用Western blot法检测Survivin过表达情况;转染细胞与牛蒡子苷元单独或联合顺铂处理,应用Annexin-Ⅴ/PI染色法检

  2. 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.

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

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    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.

  4. 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.

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

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    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.

  6. Analysis of Survivin Expression in Subtypes of Lymphoma%不同类型淋巴瘤Survivin的表达及其意义

    Institute of Scientific and Technical Information of China (English)

    顾霞; 林汉良

    2004-01-01

    背景与目的:淋巴瘤的诊断与分型是临床病理诊断的难点.本研究检测抗凋亡基因survivin在不同类型淋巴瘤中的表达,并探讨其对淋巴瘤分型的意义.方法:用免疫组化法检测83例淋巴瘤、5例淋巴结反应性增生石蜡标本中survivin蛋白的表达;同时用RT-PCR检测K562、HL-60、Raji、Jurkat细胞系和以上病例中18例淋巴瘤及2例淋巴结反应性增生新鲜标本中survivin mRNA的表达;对不同类型的淋巴瘤survivin蛋白及mRNA表达进行半定量分析.结果:Survivin蛋白在非霍奇金淋巴瘤(non-Hodgkin's lymphoma,NHL)的DLBL(diffuse large B cell lymphoma)、BL(Burkitt lymphoma)、LBL(lymphoblastic lymphoma)中有较高的表达,分别为87.2%(34/39)、100%(2/2)、85.7%(6/7),而在FL(follicularlymphoma)、MALT(extranodalmarginal zone B-cell lymphoma of mucosa-associated lymphoid tissue)和MZL(marginalzone lymphoma)中表达较低,分别为22.2%、33.3%和40.0%,且多为弱阳性.高表达组(DLBL、BL、LBL)与低表达组(FL、MZL、MALT)之间差异有统计学意义(P<0.01).并且DLBL中survivin阳性者中位年龄为57岁,明显高于阴性者41岁.霍奇金淋巴瘤(Hodgkin's lymphoma,HL)中大部分R-S(Reed-Sternberg)细胞表达survivin蛋白.NHL中survivinmRNA的检测结果与其蛋白水平呈正相关(相关系数r=0.627 0,P<0.01).结论:Survivin蛋白及mRNA表达水平在不同类型淋巴瘤存在着明显的差异,survivin可能作为一个分子标记对淋巴瘤分型具有一定的价值.

  7. Inhibitory effect of small interference RNA targeting survivin nanospheres on human pancreatic carcinoma BXPC-3 cell growth%Survivin siRNA纳米载体的制备及其对胰腺癌细胞生物学特性的影响

    Institute of Scientific and Technical Information of China (English)

    潘新亭; 朱青云; 吴力群; 赵志惠; 曹景玉; 王祖森; 胡维昱; 韩冰

    2012-01-01

    Objectives To used nanotechnology and gene interference to block survivin expression in pancreatic carcinoma ceil and to study the inhibitory effect of small interference RNA targeting survivin nanospheres on the proliferation and apoptosis of human pancreatic cancer BXPC-3 cells. Methods: Human pancreatic cancer BXPC-3 cells cultured in vitro were assigned into four groups: saline, gene-free nanospheres, siRNA-survivin and siRNA-survivin nanospheres groups. Survivin mRNA expression was detected by RT-PCR; Survivin protein expression was detected by Western blot; Apoptosis of BXPC-3 cells was determined by flow cytometry; BXPC-3 cells growth was examined by MTT. Reautlts At 72 hours after treatment, both survivin mRNA expression and survivin protein expression in the siRNA-survivin nanospheres group were significantly less than other three groups (P<0.05). BXPC-3 cell growth was remarkably inhibited in the siRNA-survivin nanospheres group, but apoptotic rate was significantly greater than other three groups (P<0.05). Conclusion* siRNA-survivin nanospheres can specifically reduce both survivin mRNA and survivin protein expressions in human pancreatic cancer BXPC-3 cells, significantly increase tumor cell apoptosis, and remarkably inhibit BXPC-3 cell proliferation.%目的:利用纳米技术和基因干扰技术设计并合成携载survivin siRNA的纳米载体,探讨survivin siRNA纳米微粒对人胰腺癌细胞BXPC-3增殖和凋亡的影响.方法:体外培养人胰腺癌BXPC-3细胞,将BXPC-3细胞随机分为4组:生理盐水组、不含基因的纳米微粒组、survivin siRNA组和survivin siRNA纳米微粒组.RT-PCR检测survivin mRNA的表达;Western blot法检测survivin蛋白的表达;流式细胞仪检测细胞凋亡情况;MTT法检测细胞增殖情况.结果:细胞培养72 h后,survivin siRNA纳米微粒组细胞的survivin mRNA和蛋白表达均低于其他3组(P<0.05).survivin siRNA纳米微粒组细胞增殖明显受抑制,生长缓慢,

  8. Calcium and survivin are involved in the induction of apoptosis by dihydroartemisinin in human lung cancer SPC-A-1 cells.

    Science.gov (United States)

    Mu, D; Chen, W; Yu, B; Zhang, C; Zhang, Y; Qi, H

    2007-01-01

    Dihydroartemisinin (DHA), a semisynthetic derivative of artemisinin isolated from the traditional Chinese herb Artemisia annua, is an effective novel antimalarial drug. Recent studies suggest that it also has anticancer effects. The present study investigated the apoptosis activity of DHA in cultured human lung cancer cells by terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assay and flow cytometry. Intracellular free calcium concentrations in the lung cancer cells were evaluated by laser scanning confocal microscopy using Fura-3/AM as probe. The observations also indicated that DHA downregulated the mRNA and protein expression level of survivin in the lung cancer cell line SPC-A-1 cells, whereas it did not affect those of caspase-4. These results demonstrated that DHA can induce apoptosis of lung cancer cell line SPC-A-1 cells and that calcium and survivin participated in the apoptotic signalling pathways. PMID:17344942

  9. Correlation between 18F-FDG PET-CT signs with the expression of Survivin and clinicopathological factors in non-small cell lung cancer tissues%非小细胞肺癌18F-FDG PET-CT影像与Survivin表达及其临床病理特征的关系

    Institute of Scientific and Technical Information of China (English)

    林珊; 韩波; 于丽娟; 王瑞芝; 宁晓明

    2011-01-01

    目的:探讨非小细胞肺癌氟-18-2-脱氧-D-葡萄糖(18F-FDG)正电子发射体层成像(PET-CT)的标准摄取值(SUV)与凋亡抑制基因Survivin表达间的关系.方法:选择行18F-FDG PET-CT扫描并经手术确诊的NSCLC患者33例,Survivin的检测,并结合临床病理特点进行统计学分析.结果:33例NSCLC病例 PET-CT的SUVmax平均为10.5±5.4,SUVmax与病理类型(鳞癌和腺癌,r=0.432,P=0.012)、临床分型(中心型和周围型,r=0.379,P=0.029)及病灶大小(r=0.743,P=0.000)具有显著的相关性;Survivin表达的阳性率为84.8%(28/33),腺癌的阳性表达高于鳞癌,u=2.29,P=0.022;SUVmax与Survivin的表达无明显相关性,与年龄、性别、病理分级、TNM分期及淋巴结转移无明显相关性.结论:PET-CT的SUVmax与组织病理类型、临床分型及病灶大小有关,Survivin的表达与组织病理类型有关,SUVmax与Survivin表达无明显相关性.%OBJECTIVE: The study focused on the correlation between PET-CT signs with the expression of Survivin (an inhibitor of apoptosis protein) and clinicopathological characteristics in nonsmall cell lung cancer (NSCLC).METHODS: Thirty-three cases of non-small cell lung cancer confirmed by pathology were examined with 2-deoxy-18 F-fluoro-D-glucose ( 18 F-FDG) PET before operation.The expression of Survivin was evaluated by immunohistochemical assay in 33 NSCLC tumor samples.The maximum standarized uptake value (SUVmax), Survivin and eight clinicopathological factors were analyzed respectively.RESULTS: The mean of SUVmax in 33 NSCLC was 10.5 ± 5.4.The correlation between SUVmax and histological type ( r= 0.432, P = 0.012), clinical type (r=0.379,P=0.029), size of lesion (r=0.743,P=0.000) were significant in the entire group; the positive expression rate of Survivin was 84.8% (28/33).It is higher in adenocarcinoma than that in squmous cell carcinoma (u= 2.29, P = 0.022); SUVmax showed no relation to the expression of Survivin, age, gender, differentiation

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

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    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.

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

    Science.gov (United States)

    Lotti, Roberta; Palazzo, Elisabetta; Petrachi, Tiziana; Dallaglio, Katiuscia; Saltari, Annalisa; Truzzi, Francesca; Quadri, Marika; Puviani, Mario; Maiorana, Antonino; Marconi, Alessandra; Pincelli, Carlo

    2016-01-01

    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 Ras(G12V)-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.

  12. Survivin-T34A: molecular mechanism and therapeutic potential

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    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

  13. Survivin is a Key Factor in the Differential Susceptibility of Gastric Endothelial and Epithelial Cells to Alcohol-Induced Injury

    OpenAIRE

    Jones, Michael K.; Padilla, Oscar R.; Zhu, Ercheng

    2010-01-01

    We previously demonstrated that the anti-apoptosis protein, survivin, plays a protective role against alcohol-induced gastric injury. Since the endothelium is a primary target of alcohol-induced gastric damage, we investigated whether survivin expression is a key factor in the greater susceptibility of gastric endothelial vs. epithelial cells to alcohol-induced injury. Here, we demonstrate that rat gastric epithelial cells (RGM1 cells, an epithelial cell line derived from normal rat gastric m...

  14. 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

  15. Survivin siRNA沉默对非小细胞肺癌细胞生物学的影响%The biological effect of Survivin on NSCLC cell line by siRNA targeting Survivin

    Institute of Scientific and Technical Information of China (English)

    翟宏芳; 崔大为; 盛树海; 孙国贵

    2014-01-01

    Objective To investigate the effect of inhibitor of apoptosis proteins (Survivin) on biological effect of human lung cancer cell line. Methods Survivin siRNA was transiently transfected into non small cell lung carcinoma (NSCLC) cell H1299 by liposome-mediated method and was detected by RT-PCR and western blot. MTT assay, cell apoptosis, cell cycle and western blot were also conducted as to the influence of the down-regulated expression of Survivin that might be found on H1299 cells biological effect. Results RT-PCR and Western blot showed that H1299 cell transfected Survivin siRNA had a lower relative expressive content than normal cell (t=10.970, P=0.000; t=9.192, P=0.003). MTT assay, cell apoptosis, cell cycles demonstrated that H1299 cell transfected Survivin siRNA had a lower survival fraction, higher cell apoptosis, more percentage of the G2/M phases (t=8.162, P=0.001;t=10.800, P=0.000). Western blot showed that the relative content of Caspase-9 protein was increased in H1299 cell transfected Survivin siRNA compared with H1299 cell untransfected Survivin (t=9.192, P=0.003). Conclusion Survivin may involved in the biological processes of lung cancer cell proliferation, apoptosis, and cell cycle.%目的:探讨凋亡抑制蛋白(Survivin)对非小细胞肺癌(NSCLC)H1299细胞株的生物学影响。方法通过脂质体介导将 Survivin 基因 siRNA 瞬时转染 H1299细胞。采用 RT-PCR及Western blot 检测转染后H1299细胞内Survivin 基因的表达水平。MTT比色法、流式细胞术(FCM)及Western blot观察细胞增殖、凋亡、周期及Caspase-9蛋白表达变化情况。结果 RT-PCR及Western blot证实,siRNA沉默的H1299细胞中Survivin mRNA、蛋白的表达水平较未转染组明显降低(t=10.970,P=0.000;t=9.192,P=0.003)。MTT、细胞凋亡及周期结果表明,siRNA 沉默的 H1299细胞其增殖能力明显减弱、细胞凋亡明显升高,G2/M期阻滞,Caspase-9蛋白表达增加(t=8

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

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    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.

  17. 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.

  18. Dimethoxy Curcumin Induces Apoptosis by Suppressing Survivin and Inhibits Invasion by Enhancing E-Cadherin in Colon Cancer Cells.

    Science.gov (United States)

    Chen, Dong; Dai, Fang; Chen, Zhehang; Wang, Saisai; Cheng, Xiaobin; Sheng, Qinsong; Lin, Jianjiang; Chen, Wenbin

    2016-01-01

    BACKGROUND Dimethoxy curcumin (DMC) is a kind of lipophilic analog of curcumin with great improvement in chemical and metabolic stability. DMC has been studied in breast and renal cancer, but no research in colon cancer has been found yet. MATERIAL AND METHODS Two colon cancer cells (HT-29 and SW480) and one normal human colon mucosal epithelial cell (NCM460) were used in this study. We studied the effect of DMC on the proliferation in vitro and in vivo. Transwell migration assay was used to estimate the inhibition of DMC on invasion. Moreover, the expressions of PARP, caspase-3, survivin and E-cadherin were detected to uncover the related signaling pathways by western blotting assay both in vitro and in vivo. RESULTS DMC significantly inhibited the growth of colon cancer cells in dose-dependent manner; IC50 for DMC was calculated to be 43.4, 28.2 and 454.8µM on HT-29, SW480 and NCM460. DMC significantly increased the apoptosis in both HT-29 (p=0.0051) and SW480 (p=0.0013) cells in vitro, and significantly suppressed the growth of both cell lines in vivo. Moreover, DMC reduced the number of migrated cells in both HT-29 (p=0.007) and SW480 (p=0.004) cells. By western blotting analysis, the cleavage of pro-caspases-3 and PARP were clearly induced by DMC to their active form, while the expression of survivin was reduced and E-cadherin was enhanced in both cells in vitro and in vivo. CONCLUSIONS DMC may exert an effective anti-tumor effect in colon cancer cells by down-regulating survivin and upregulating E-cadherin. PMID:27614381

  19. 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%

  20. Polymeric micelles containing reversibly phospholipid-modified anti-survivin siRNA: a promising strategy to overcome drug resistance in cancer

    OpenAIRE

    Salzano, G; Riehle, R.; Navarro, Gemma; Perche, Federico; Rosa, G.; Torchilin, VT

    2013-01-01

    The discovery that survivin, a small anti-apoptotic protein, is involved in chemoresistance, opens a new scenario to overcome the drug resistance in cancer. It was shown that siRNA can efficiently inhibit the expression of survivin in cancer cells. However, the clinical use of siRNA is still hampered by an unfavorable pharmacokinetic profile. To address this problem, earlier we developed a novel system to deliver siRNA into cancer cells. Namely, we reversibly modified the survivin siRNA with ...

  1. 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.

  2. An efficient antigene activity and antiproliferative effect by targeting the Bcl-2 or survivin gene with triplex forming oligonucleotides containing a W-shaped nucleoside analogue (WNA-βT).

    Science.gov (United States)

    Taniguchi, Yosuke; Sasaki, Shigeki

    2012-10-01

    Triplex forming oligonucleotides (TFOs) are some of the most promising tools in the antigene strategy for the development of gene targeting therapeutics. However, the stable triplex formation is restricted to the homopurine sequences consisting of purine nucleosides, dG and dA. Therefore, the T or dC nucleoside in the homopurine strand inhibits the stable triplex formation. We have developed W-shaped nucleoside analogues (WNAs) for the formation of the unnatural type triplex DNA, with sequences containing the interrupting site in an antiparallel triplex formation. In the present study, we tested the antigene effect of TFOs having WNA-βT, which increased the stability of the triplex formation with a target sequence including the TA interrupting site. We designed the GU TFO (WNA) and GU TFO (natural) for targeting sequences of the Bcl-2 or survivin oncogene. The gel shift assay showed that the TFO (WNA) formed more stable triplexes than the natural TFO. Remarkably, the Bcl-2- or survivin-targeted TFO (WNA) inhibited the cell proliferation and induced a caspase-dependent apoptosis. It was confirmed that the survivin-targeted TFO (WNA) more effectively decreased the number of survivin products in the A549 cell than the natural TFOs. PMID:22987068

  3. 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.

  4. Localization and upregulation of survivin in cancer health disparities: a clinical perspective

    Directory of Open Access Journals (Sweden)

    Khan S

    2015-07-01

    Full Text Available Salma Khan,1,2 Heather Ferguson Bennit,1,2 Malyn May Asuncion Valenzuela,1,2 David Turay,1,3 Carlos J Diaz Osterman,1,2 Ron B Moyron,1,2 Grace E Esebanmen,1,2 Arjun Ashok,1,2 Nathan R Wall1,2 1Department of Biochemistry, 2Center for Health Disparities and Molecular Medicine, 3Department of Anatomy, Loma Linda University School of Medicine, Loma Linda, CA, USA Abstract: Survivin is one of the most important members of the inhibitors of apoptosis protein family, as it is expressed in most human cancers but is absent in normal, differentiated tissues. Lending to its importance, survivin has proven associations with apoptosis and cell cycle control, and has more recently been shown to modulate the tumor microenvironment and immune evasion as a result of its extracellular localization. Upregulation of survivin has been found in many cancers including breast, prostate, pancreatic, and hematological malignancies, and it may prove to be associated with the advanced presentation, poorer prognosis, and lower survival rates observed in ethnically diverse populations. Keywords: survivin, cancer, exosomes, health disparity

  5. 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.

  6. 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

  7. 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

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

    International Nuclear Information System (INIS)

    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. 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. 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). Using the same threshold as our previous study, we have validated survivin CNR as a marker of good prognosis in

  9. 阿米福汀对HL-60细胞中survivin表达的影响%Effect of amifostine on survivin expression in HL-60 cells in vitro

    Institute of Scientific and Technical Information of China (English)

    武婕萍; 马梁明; 周永安

    2007-01-01

    目的 探讨阿米福汀(WR-2721,amifostine,AMI)对人髓系白血病细胞HL-60中survivin表达的影响.方法 不同浓度AMI(1,10,1 000 μmol/L)干预HL-60细胞24,48,72 h,用MTT比色法检测细胞的生长抑制作用;用半定量RT-PCR方法检测抑凋亡基因survivin mRNA的表达水平.结果 阿米福汀可显著抑制HL-60细胞的增殖和下调抑凋亡基因survivin 的表达水平,且均有明显浓度和时间依赖性.结论 AMI可能通过下调抑凋亡基因survivin的表达,解除其抑制凋亡效应,从而抑制HL-60细胞的增殖.

  10. 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.

  11. 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. PMID:26708213

  12. 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

  13. 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.

  14. 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

  15. Survivin表达对鼠髓样树突状细胞成熟的影响%Effect of survivin expression on the maturation of mouse marrow dendritic cells

    Institute of Scientific and Technical Information of China (English)

    黎衍敏; 苟欣; 何卫阳; 王海峰

    2009-01-01

    目的:探讨survivin的表达变化对小鼠骨髓来源树突状细胞分化成熟的影响.方法:无菌条件下取小鼠的骨髓细胞作为树突状前体细胞,用粒细胞巨噬细胞集落刺激因子(Granulocyte macrophage colony-stimulating factor,GM-CSF)和白细胞介素4(Interleukin 4,IL-4)的培养基定向诱导树突状前体细胞分化成未成熟树突状细胞(Immature dendritic cells,iDC),用GM-CSF,IL-4及肿瘤坏死因子-α(Tumor necrosis factor-alpha,TNF-α)定向诱导树突状前体细胞分化成成熟树突状细胞(Mature dendritic cells,mDC):运用半定量RT-PCR,Western blot比较树突状前体细胞、iDC与mDC中survivin mRNA、survivin蛋白的含量变化.结果:RT-PCR检测结果显示,树突状前体细胞survivin mRNA表达值为(0.735±0.081),iDC为(0.596±0.073,P<0.05),mDC为(0.393±0.052,P<0.01);Western blot检测结果显示:树突状前体细胞survivin蛋白的表达值为(0.767±0.045),iDC的表达值为(0.396±0.016,P<0.05),mDC的表达值为(0.168±0.009,P<0.01).结论:survivin在树突状前体细胞中高表达,在iDC与mDC中表达量略低;且survivin在iDC中的表达量高于mDC,这个结论为我们的后续实验提供了理论基础.

  16. 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.

  17. 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.

  18. 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.

  19. YAP promotes malignant progression of Lkb1-deficient lung adenocarcinoma through downstream regulation of survivin.

    Science.gov (United States)

    Zhang, Wenjing; Gao, Yijun; Li, Fuming; Tong, Xinyuan; Ren, Yan; Han, Xiangkun; Yao, Shun; Long, Fei; Yang, Zhongzhou; Fan, Hengyu; Zhang, Lei; Ji, Hongbin

    2015-11-01

    The serine/threonine kinase LKB1 is a well-characterized tumor suppressor that governs diverse cellular processes, including growth, polarity, and metabolism. Somatic-inactivating mutations in LKB1 are observed in about 15% to 30% of non-small cell lung cancers (NSCLC). LKB1 inactivation confers lung adenocarcinomas (ADC) with malignant features that remain refractory to therapeutic intervention. YAP activation has been linked to LKB1 deficiency, but the role of YAP in lung ADC formation and progression is uncertain. In this study, we showed that ectopic expression of YAP in type II alveolar epithelial cells led to hyperplasia in mouse lungs. YAP overexpression in the Kras(G12D) lung cancer mouse model accelerated lung ADC progression. Conversely, YAP deletion dramatically delayed the progression of lung ADC in LKB1-deficient Kras(G12D) mice. Mechanistic studies identified the antiapoptotic oncoprotein survivin as the downstream mediator of YAP responsible for promoting malignant progression of LKB1-deficient lung ADC. Collectively, our findings identify YAP as an important contributor to lung cancer progression, rationalizing YAP inhibition in the context of LKB1 deficiency as a therapeutic strategy to treat lung ADC.

  20. 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.

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

    International Nuclear Information System (INIS)

    Highlights: → Intrinsic trastuzumab resistance occurs in ∼70% of metastatic HER2 + breast carcinomas (BC). → Approximately 15% of early HER2 + BC relapse in spite of treatment with trastuzumab-based therapies. → HER2-independent downstream pro-survival pathways might underlie trastuzumab refractoriness. → Survivin is indispensable for proliferation and survival of HER2 + BC unresponsive to HER2-targeted therapies ab initio. → 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 IC50 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 ∼4 times more survivin protein than trastuzumab-responsive SKBR3 cells. In response to trastuzumab, JIMT-1 cells accumulated ∼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 suppressed in JIMT-1 cells. Our current findings may

  2. Silencing of signal transducer and activator of transcription 3 expression by RNA interference suppresses growth of human hepatocellular carcinoma in tumor-bearing nude mice

    Institute of Scientific and Technical Information of China (English)

    Jing Li; Yun-Feng Piao; Zheng Jiang; Li Chen; Hai-Bo Sun

    2009-01-01

    AIM: To explore the effect of silencing of signal transducer and activator of transcription 3 (STAT3) expression by RNA interference (RNAi) on growth of human hepatocellular carcinoma (HCC) in tumorbearing nude mice in vivo.METHODS: To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP (pSH1-siRNASTAT3) and establish the tumor-bearing nude mouse model of the HCC cell line SMMC7721, we used intratumoral injection together with electroblotting to transfect the recombinant plasmid pSH1-siRNASTAT3 into the transplanted tumor. The weight of the nude mice and tumor volumes were recorded. STAT3 gene transcription was detected by semi-quantitative reverse transcription polymerase chain reaction (RTPCR).Level of protein expression and location of STAT3 were determined by Western blotting and immunohistochemical staining. STAT3-related genes such as survivin, c-myc, VEGF, p53 and caspase3 mRNA and protein expression were detected in tumor tissues at the same time. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptosis of tumor cells.RESULTS: The weight of the treated nude mice increased, and the tumor volume decreased markedly compared with those of the mock-treated and negative control groups ( P < 0.01). The results of RT-PCR and Western blotting showed that mRNA and protein levels of STAT3 declined markedly in the treated group. The change in STAT3-related gene expression in tumor tissues at the mRNA and protein level also varied, the expression of survivin, VEGF and c-myc were obviously reduced, and expression of p53 and caspase3 increased ( P < 0.01). Most of the tumor tissue cells in the treated group developed apoptosis that was detected by TUNEL assay.CONCLUSION: Silencing of STAT3 expression by RNAi significantly inhibits expression of STAT3 mRNA and protein, and suppresses growth of human HCC in tumor-bearing nude mice. The mechanism may be related to down-regulation of survivin, VEGF

  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. Survivin Expression in Medulloblastoma and its Clinical Significance%survivin蛋白在髓母细胞瘤中的表达及其与预后的关系

    Institute of Scientific and Technical Information of China (English)

    王多彬; 蒋永明; 吕明学; 曾春; 康东; 刘翼; 张跃康

    2009-01-01

    目的:通过检测髓母细胞瘤(Medulloblastoma)中凋亡抑制蛋白(survivin)的表达情况,探讨它的临床意义,为髓母细胞瘤的治疗寻求新的思路.方法:对50例髓母细胞瘤及10例正常脑组织石蜡切片行免疫组化检测,通过对survivin蛋白在髓母细胞瘤患者与正常脑组织中的表达差异,并运用相关统计分析方法分析它在不同性别、年龄段、肿瘤大小、病理类型、临床分期的表达差异及其与预后的相关性.结果:(1)髓母细胞瘤中survivin蛋白阳性表达率为72%,明显高于正常脑组织的阳性表达率0%,二者表达差异明显(P0.05),而年龄及TM临床分期在两组间差异显著(P分别为0.044和0.027).(2)survivin阳性患者预后与和阴性组相比差异显著(P=0.001).结论:survivin可能参与了对髓母细胞瘤细胞凋亡的调控,并对髓母细胞瘤的发生发展可能起着重要作用.由于survivin蛋白表达与髓母细胞瘤患者年龄、临床分期及预后等也密切相关,可作为判断病情和评价预后的指标.

  5. Effect of NF-κB, survivin, Bcl-2 and Caspase3 on apoptosis of gastric cancer cells induced by tumor necrosis factor related apoptosis inducing ligand

    Institute of Scientific and Technical Information of China (English)

    Liu-Qin Yang; Dian-Chun Fang; Rong-Quan Wang; Shi-Ming Yang

    2004-01-01

    AIM: To study the effect of NF-κB, survivin, Bcl-2 and Caspase3 on tumor necrosis factors related apoptosis inducing ligand (TRAIL) induced apoptosis of gastric cancer cells.METHODS: Gastric cancer cells of SGC-7901, MKN28,MKN45 and AGS lines were cultured in PRMI-1640 medium and the apoptosis rates of the cells of 4 lines were observed after treatment of tumor necrosis factors related apoptosis inducing ligand (TRAIL) with a flow cytometer. The expression of NF-κB, survivin, Bcl-2 and Caspase3 in gastric cancer cells of 4 lines was analyzed with Western blot.RESULTS: After the gastric cancer cells were exposed to TRAIL 300 ng/ml for 24 hours, the apoptosis rate was 36.05%, 20.27%, 16.50% and 11.80% in MKN28, MKN45,AGS and SGC-7901cells respectively. Western blot revealed that the expressions of NF-κB and survivin were lower in MKN28 cells than in MKN45, AGS and SGC-7901 cells. In contrast, the expression of Caspase3 was higher in MKN28 cells than in MKN45, AGS and SGC-7901 cells.CONCLUSION: There is a selectivity of TRAIL potency to induce apoptosis in gastric cancer cells of different cell lines.The anticancer potency of TRAIL is associated with the decreased expression of NF-κB and survivin and increased expression of Caspase3 of gastric cancer cells.

  6. Synthesis and evaluation of a radioiodinated 4,6-diaryl-3-cyano-2-pyridinone derivative as a survivin targeting SPECT probe for tumor imaging.

    Science.gov (United States)

    Fuchigami, Takeshi; Mizoguchi, Tatsuya; Ishikawa, Natsumi; Haratake, Mamoru; Yoshida, Sakura; Magata, Yasuhiro; Nakayama, Morio

    2016-02-01

    Survivin is overexpressed in most of the cancerous tissues but not in terminally differentiated normal tissues, making it an attractive target for diagnosis and therapy of various types of cancers. In this study, we aimed to develop 4,6-diaryl-3-cyano-2-pyridinone (DCP) derivatives, as novel cancer imaging probes that target survivin. Chloro and iodo analogs of DCP (CDCP and IDCP, respectively) were successfully synthesized by using a previously unreported carbon monoxide-free procedure. IDCP exhibited a slightly higher binding affinity for recombinant human survivin (Kd=34 nM) than that of CDCP (Kd=44 nM). Fluorescence staining indicated that both CDCP and IDCP showed high signals in MDA-MB-231 cells with high levels of survivin expression. Significantly low fluorescent signals were observed in MCF-10A cells, which showed low levels of survivin expression. [(125)I]IDCP was synthesized for the application of IDCP to single photon emission computed tomography (SPECT) imaging. Quantitative in vitro binding of [(125)I]IDCP in cell cultures showed results consistent to those observed after fluorescent staining. In vivo biodistribution studies in tumor-bearing mice demonstrated that the tumor uptake of [(125)I]IDCP increased gradually with time and was 0.65% injected dose per gram (% ID/g) at 180 min. The maximum tumor/blood and tumor/muscle ratio at 60 min were 0.87 and 2.27, respectively, indicating inadequate [(125)I]IDCP accumulation in tumors necessary for in vivo imaging. Although further structural modifications are necessary to improve pharmacokinetic properties of IDCP, this study demonstrates the feasibility of using the DCP backbone as a scaffold for the development of survivin-targeting tumor imaging probes. PMID:26733475

  7. Survivin upregulation, dependent on leptin-EGFR-Notch1 axis, is essential for leptin induced migration of breast carcinoma cells

    OpenAIRE

    Knight, Brandi B.; Oprea-Ilies, Gabriela M.; Nagalingam, Arumugam; Yang, Lily; Cohen, Cynthia; Saxena, Neeraj K; Sharma, Dipali

    2011-01-01

    Obese breast cancer patients exhibit a higher risk for larger tumor burden and increased metastasis. Molecular effects of obesity on carcinogenesis are mediated by autocrine and paracrine effects of adipocytokine leptin. Leptin participates in tumor progression and metastasis of human breast. We show that leptin induces clonogenicity and migration potential of breast cancer cells. We found that survivin expression is induced in response to leptin. In this study, we examine the role and leptin...

  8. 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.

  9. 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.

  10. 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

  11. Enhancing glioblastoma cell sensitivity to chemotherapeutics: A strategy involving survivin gene silencing mediated by gemini surfactant-based complexes.

    Science.gov (United States)

    Cruz, Rita Q; Morais, Catarina M; Cardoso, Ana M; Silva, Sandra G; Vale, Maria L; Marques, Eduardo F; Pedroso de Lima, Maria C; Jurado, Amália S

    2016-07-01

    Glioblastoma (GBM), the highest grade astrocytoma, is one of the most aggressive and challenging cancers to treat. The standard treatment is usually limited due to the intrinsic resistance of GBM to chemotherapy and drug non-specific effects. Therefore, new therapeutic strategies need to be developed to target tumor cells, sparing healthy tissues. In this context, the inhibitor-of-apoptosis protein (IAP) survivin emerges as an ideal target for a gene silencing approach, since it is sharply differentially expressed in cancer tissues. In this work, two different families of cationic gemini surfactants (bis-quat conventional and serine-derived) were tested regarding their efficiency to deliver small interfering RNAs (siRNAs) in a human GBM cell line (U87), in order to select an effective siRNA anti-survivin carrier. Importantly, survivin downregulation combined with administration of the chemotherapeutic agents temozolomide or etoposide resulted in a synergistic cytotoxic effect, thus revealing to be a promising strategy to reduce the chemotherapeutic doses for GBM treatment. PMID:27106606

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  14. Simultaneous gene silencing of Bcl-2, XIAP and Survivin re-sensitizes pancreatic cancer cells towards apoptosis

    International Nuclear Information System (INIS)

    Pancreatic ductal adenocarcinoma shows a distinct apoptosis resistance, which contributes significantly to the aggressive nature of this tumor and constrains the effectiveness of new therapeutic strategies. Apoptosis resistance is determined by the net balance of the cells pro-and anti-apoptotic 'control mechanisms'. Numerous dysregulated anti-apoptotic genes have been identified in pancreatic cancer and seem to contribute to the high anti-apoptotic buffering capacity. We aimed to compare the benefit of simultaneous gene silencing (SGS) of several candidate genes with conventional gene silencing of single genes. From literature search we identified the anti-apoptotic genes XIAP, Survivin and Bcl-2 as commonly upregulated in pancreatic cancer. We performed SGS and silencing of single candidate genes using siRNA molecules in two pancreatic cancer cell lines. Effectiveness of SGS was assessed by qRT-PCR and western blotting. Apoptosis induction was measured by flow cytometry and caspase activation. Simultaneous gene silencing reduced expression of the three target genes effectively. Compared to silencing of a single target or control, SGS of these genes resulted in a significant higher induction of apoptosis in pancreatic cancer cells. In the present study we performed a subliminal silencing of different anti-apoptotic target genes simultaneously. Compared to silencing of single target genes, SGS had a significant higher impact on apoptosis induction in pancreatic cancer cells. Thereby, we give further evidence for the concept of an anti-apoptotic buffering capacity of pancreatic cancer cells

  15. Serum Survivin Levels and Outcome of Chemotherapy in Patients with Malignant Mesothelioma

    Directory of Open Access Journals (Sweden)

    Katja Goričar

    2015-01-01

    Full Text Available Background. Survivin is an inhibitor of apoptosis protein involved in the regulation of cell proliferation that could be used as a marker for cancer diagnosis or prognosis. Our aim was to evaluate whether serum survivin levels influence the outcome of cisplatin-based chemotherapy in patients with malignant mesothelioma (MM. Methods. Serum survivin levels were determined using human survivin enzyme-linked immunosorbent assay in 78 MM patients before chemotherapy, after chemotherapy, and at disease progression. The influence on tumor response and survival was evaluated using nonparametric tests and Cox regression. Results. A median serum survivin level at diagnosis was 4.1 (0–217.5 pg/mL. Patients with a progressive disease had significantly higher survivin levels before chemotherapy (p = 0.041. A median serum survivin level after chemotherapy was 73.1 (0–346.2 pg/mL. If survivin levels increased after chemotherapy, patients had, conversely, better response (p = 0.001, OR = 5.40, 95% CI = 1.98–14.72. Unexpectedly, patients with increased survivin levels after chemotherapy also had longer progression-free (p < 0.001, HR = 0.33, 95% CI = 0.20–0.57 and overall survival (p = 0.001, HR = 0.29, 95% CI = 0.14–0.58. Conclusions. These results suggest that serum survivin levels before and during chemotherapy could serve as a biomarker predicting MM treatment response.

  16. 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.

  17. Effect of Survivin gene on proliferation inhibition and apoptosis of SiHa cells after RNA interference%RNA干涉Survivin基因对SiHa细胞增殖抑制及凋亡的影响

    Institute of Scientific and Technical Information of China (English)

    赵丽晶; 徐红梅; 刘艳波; 梁作文; 许多; 滕菲; 赵秀云; 赵淑华; 赵雪俭

    2011-01-01

    Objective: To study the effect of Survivin gene on proliferation inhibition and apoptosis of SiHa cells after RNA interference and the mechanism.Methods: Recombinant plasmid (pU6 -siRNA -Survivin ) and empty plasmid were used to transfect SiHa cells, MTT method was used to detect cell proliferation and transcription level of Survivin gene, acridine orange staining and flow cytometry were used to detect cell apoptosis, immunohistochemical staining was used to detect the expressions of Survivin and Caspase 3.Results: After RNA interference, the proliferation of SiHa cells was inhibited, and apoptosis appeared, the transcription of Survivin gene was down -regulated, while the transcription of Caspase 3 was up - regulated.Conclusion: Survivin gene may induce the proliferation inhibition and apoptosis of SiHa cells after RNA interference, the mechanism is related to up - regulation of Caspase 3.%目的:研究RNA干涉survivin基因对宫颈癌SiHa细胞的增殖抑制及凋亡的影响并探讨其作用机制.方法:重组质粒(pU6-siRNA-Survivin)及空质粒转染SiHa细胞,MTT法检测细胞增殖情况,检测survivin基因转录水平,吖啶噔染色、流式细胞术检测细胞凋亡,免疫组化染色检测survivin、caspase3基因表达.结果:RNA干涉后,抑制SiHa细胞增殖并出现细胞凋亡,survivin基因转录表达下调,caspase3基因表达上调.结论:RNA干涉survivin基因可导致SiHa细胞增殖抑制并凋亡,其机制与caspase3基因表达上调有关.

  18. 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

  19. Effects of all-trans-retinoic add on chemotherapeutics sensibility and Survivin gene expression of human colon cancer LoVo cell line%全反式维甲酸对人结肠癌LoVo细胞化疗药物敏感性及Survivin基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    胡宝光; 郑宗珩; 卫洪波; 韩晓燕; 魏波; 黄江龙

    2011-01-01

    Objective To study the effects of all-trans-retinoic acid (ATRA) on chemotherapeutics sensibility and expression of Survivin in human LoVo colon cancer cell line, and to provide a theoretical basis for clinical application of ATRA.Methods Flow cytometer (FCM) was used to detect the cell cycle of LoVo colon cancer cell line after treated with various dose of ATRA (10-5 mol/L, 10-6 mol/L and 10-7 mol/L)for 24 h, 48 h, 72 h, 5 d, 9 d and 15 d respectively.The cells were divided into ATRA group (10-6 mol/L)and control group (cultured in routine medium RPMI1640).Meanwhile, each group were divided into subgroups respectively treated with 0 g/L, 2 g/L, 4 g/L and 6 g/L 5-fluorouracil(5-FU) , or with 0 mg/L, 200 mg/L, 400 mg/L and 600 mg/L mitomycin (MMC).MTT assay was used to detect the viability of cells so as to analyze the change in chemotherapeutics sensibility.Drug interactions were analyzed according to the interaction effects.Staining with acridine orange (AO) and ethidium bromide (EB) was used to study the apoptosis in control group(RPMI1640) , ATRA group (10-6 mol/L), 5-FU group (4 g/L), MMC group (400 mg/L), ATRA (10-6 mol/L) + 5-FU (4 g/L) group and ATRA (10-6 mol/L) + MMC (400 mg/L) group.The expression of Survivin in the LoVo cell line was investigated by immunofluorescence technique.Results Compared with the control group, the percentage of cell in Go-G1 phase increased after interference with various doses of ATRA for 24 hours, peaked at 48 h, and gradually decreased afterwards, while the percentage of cells in stages S and G2-M decreased, reach the trough at 48 h, and then increased gradually.Effects of 10-6 mol/L ATRA were most significant at all time spots.Compared with control group, 10-6 mol/L ATRA reduced cell survival rate (P<0.05),and the survival rates in control subgroups treated with various doses of 5-FU were significantly higher than those in ATRA group (all P<0.05).In MMC-treated control group, cell survival rate with 200 mg/L MMC was lower

  20. Endoglin regulates cyclooxygenase-2 expression and activity.

    Science.gov (United States)

    Jerkic, Mirjana; Rivas-Elena, Juan V; Santibanez, Juan F; Prieto, Marta; Rodríguez-Barbero, Alicia; Perez-Barriocanal, Fernando; Pericacho, Miguel; Arévalo, Miguel; Vary, Calvin P H; Letarte, Michelle; Bernabeu, Carmelo; López-Novoa, Jose M

    2006-08-01

    The endoglin heterozygous (Eng(+/-)) mouse, which serves as a model of hereditary hemorrhagic telangiectasia (HHT), was shown to express reduced levels of endothelial NO synthase (eNOS) with impaired activity. Because of intricate changes in vasomotor function in the Eng(+/-) mice and the potential interactions between the NO- and prostaglandin-producing pathways, we assessed the expression and function of cyclooxygenase (COX) isoforms. A specific upregulation of COX-2 in the vascular endothelium and increased urinary excretion of prostaglandin E(2) were observed in the Eng(+/-) mice. Specific COX-2 inhibition with parecoxib transiently increased arterial pressure in Eng(+/-) but not in Eng(+/+) mice. Transfection of endoglin in L6E9 myoblasts, shown previously to stimulate eNOS expression, led to downregulation of COX-2 with no change in COX-1. In addition, COX-2 promoter activity and protein levels were inversely correlated with endoglin levels, in doxycyclin-inducible endothelial cells. Chronic NO synthesis inhibition with N(omega)-nitro-l-arginine methyl ester induced a marked increase in COX-2 only in the normal Eng(+/+) mice. N(omega)-nitro-l-arginine methyl ester also increased COX-2 expression and promoter activity in doxycyclin-inducible endoglin expressing endothelial cells, but not in control cells. The level of COX-2 expression following transforming growth factor-beta1 treatment was less in endoglin than in mock transfected L6E9 myoblasts and was higher in human endothelial cells silenced for endoglin expression. Our results indicate that endoglin is involved in the regulation of COX-2 activity. Furthermore, reduced endoglin levels and associated impaired NO production may be responsible, at least in part, for augmented COX-2 expression and activity in the Eng(+/-) mice. PMID:16840721

  1. Analysis of Survivin-Specific T Cells in Breast Cancer Patients Using Human DCs Engineered with Survivin mRNA

    DEFF Research Database (Denmark)

    Met, Ozcan; Svane, Inge Marie

    2013-01-01

    The observation that dendritic cells (DCs) charged with tumor-associated antigens (TAAs) is a potent strategy to elicit protective immunity in tumor-bearings hosts has prompted extensive testing of DCs as cellular adjuvant in cancer vaccines. To improve the clinical development of DC-based cancer...... vaccines, it may be beneficial to analyze preexistent immunity against TAAs in cancer patients because it may be easier to expand a memory pool of T cells compared to generating new immunity. Recent research shows that engineering DCs to synthesize tumor epitopes endogenously by transfecting DCs with m......RNA-encoding TAAs are particular effective in stimulating robust T-responses in vitro and in vivo. In this chapter, we describe the methodology to analyze for survivin-specific T cells in breast cancer patients using human DCs engineered with survivin mRNA....

  2. 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...... expression were significantly associated with breast cancer grade. There was a significant association between increased CIN and both increased AURKA and SURVIVIN expression. AURKA gene amplification was also associated with increased CIN. To our knowledge this is the largest study assessing CIN status...... in parallel with the expression of the mitotic regulators AURKA and SURVIVIN. These data suggest that elevated expression of AURKA and SURVIVIN, together with AURKA gene amplification, are associated with increased CIN in breast cancer, and may be used as a proxy for CIN in breast cancer samples...

  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

    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...... 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...... independent prognostic factors for response and survival after cisplatin-containing chemotherapy in patients with advanced bladder cancer....

  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

    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...... 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...... independent prognostic factors for response and survival after cisplatin-containing chemotherapy in patients with advanced bladder cancer....

  5. 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.

  6. CRM1-mediated nuclear export determines the cytoplasmic localization of the antiapoptotic protein survivin

    NARCIS (Netherlands)

    Rodriguez, JA; Span, SW; Ferreira, CGM; Kruyt, FAE; Giaccone, G

    2002-01-01

    Survivin is a member of the inhibitor of apoptosis (IAP) family of negative regulators of programmed cell death that is frequently overexpressed in human tumors. Survivin is not only involved in the regulation of apoptosis, but is also known to play a role in the control of cell cycle progression at

  7. The novel curcumin analog FLLL32 decreases STAT3 DNA binding activity and expression, and induces apoptosis in osteosarcoma cell lines

    International Nuclear Information System (INIS)

    Curcumin is a naturally occurring phenolic compound shown to have a wide variety of antitumor activities; however, it does not attain sufficient blood levels to do so when ingested. Using structure-based design, a novel compound, FLLL32, was generated from curcumin. FLLL32 possesses superior biochemical properties and more specifically targets STAT3, a transcription factor important in tumor cell survival, proliferation, metastasis, and chemotherapy resistance. In our previous work, we found that several canine and human osteosarcoma (OSA) cell lines, but not normal osteoblasts, exhibit constitutive phosphorylation of STAT3. Compared to curcumin, we hypothesized that FLLL32 would be more efficient at inhibiting STAT3 function in OSA cells and that this would result in enhanced downregulation of STAT3 transcriptional targets and subsequent death of OSA cells. Human and canine OSA cells were treated with vehicle, curcumin, or FLLL32 and the effects on proliferation (CyQUANT®), apoptosis (SensoLyte® Homogeneous AMC Caspase- 3/7 Assay kit, western blotting), STAT3 DNA binding (EMSA), and vascular endothelial growth factor (VEGF), survivin, and matrix metalloproteinase-2 (MMP2) expression (RT-PCR, western blotting) were measured. STAT3 expression was measured by RT-PCR, qRT- PCR, and western blotting. Our data showed that FLLL32 decreased STAT3 DNA binding by EMSA. FLLL32 promoted loss of cell proliferation at lower concentrations than curcumin leading to caspase-3- dependent apoptosis, as evidenced by PARP cleavage and increased caspase 3/7 activity; this could be inhibited by treatment with the pan-caspase inhibitor Z-VAD-FMK. Treatment of OSA cells with FLLL32 decreased expression of survivin, VEGF, and MMP2 at both mRNA and protein levels with concurrent decreases in phosphorylated and total STAT3; this loss of total STAT3 occurred, in part, via the ubiquitin-proteasome pathway. These data demonstrate that the novel curcumin analog FLLL32 has biologic activity

  8. Hormone activation of baculovirus expressed progesterone receptors.

    Science.gov (United States)

    Elliston, J F; Beekman, J M; Tsai, S Y; O'Malley, B W; Tsai, M J

    1992-03-15

    Human and chicken progesterone receptors (A form) were overproduced in a baculovirus expression system. These recombinant progesterone receptors were full-length bound progesterone specifically and were recognized by monoclonal antibodies, AB52 and PR22, specific for human and chicken progesterone receptor, respectively. In gel retardation studies, binding of recombinant human and chicken progesterone receptors to their progesterone response element (PRE) was specific and was enhanced in the presence of progesterone. Binding of human progesterone receptor to the PRE was also enhanced in the presence of the antiprogestin, RU486, but very little effect was observed in the presence of estradiol, dexamethasone, testosterone, and vitamin D. In our cell-free transcription system, human progesterone receptor induced transcription in a receptor-dependent and hormone-activable manner. Receptor-stimulated transcription required the presence of the PRE in the test template and could be specifically inhibited by excess PRE oligonucleotides. Furthermore, chicken progesterone receptor also induced in vitro transcription in a hormone-activable manner. These results demonstrate that steroid receptors overexpressed in a baculovirus expression system are functional and exhibit steroid-responsive binding and transcription. These observations support our present understanding of the mechanism of steroid receptor-regulated gene expression and provide a technological format for studies of the role of hormone and antihormone in altering gene expression. PMID:1544902

  9. 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).

  10. Differential Control of Growth, Apoptotic Activity, and Gene Expression in Human Breast Cancer Cells by Extracts Derived from Medicinal Herbs Zingiber officinale

    Directory of Open Access Journals (Sweden)

    Ayman I. Elkady

    2012-01-01

    Full Text Available The present study aimed to examine the antiproliferative potentiality of an extract derived from the medicinal plant ginger (Zingiber officinale on growth of breast cancer cells. Ginger treatment suppressed the proliferation and colony formation in breast cancer cell lines, MCF-7 and MDA-MB-231. Meanwhile, it did not significantly affect viability of nontumorigenic normal mammary epithelial cell line (MCF-10A. Treatment of MCF-7 and MDA-MB-231 with ginger resulted in sequences of events marked by apoptosis, accompanied by loss of cell viability, chromatin condensation, DNA fragmentation, activation of caspase 3, and cleavage of poly(ADP-ribose polymerase. At the molecular level, the apoptotic cell death mediated by ginger could be attributed in part to upregulation of Bax and downregulation of Bcl-2 proteins. Ginger treatment downregulated expression of prosurvival genes, such as NF-κB, Bcl-X, Mcl-1, and Survivin, and cell cycle-regulating proteins, including cyclin D1 and cyclin-dependent kinase-4 (CDK-4. On the other hand, it increased expression of CDK inhibitor, p21. It also inhibited the expression of the two prominent molecular targets of cancer, c-Myc and the human telomerase reverse transcriptase (hTERT. These findings suggested that the ginger may be a promising candidate for the treatment of breast carcinomas.

  11. survivin与宫颈鳞癌新辅助化疗和放疗敏感性的相关性研究%Correlative Study between Expression of the Survivin and the Neoadjuvant Chemotherapy and Radiosensitivity in Cervical Squamous Cell Carcinoma

    Institute of Scientific and Technical Information of China (English)

    高碧燕; 张明; 杨毅; 吴星娆; 金丛国

    2010-01-01

    目的 探讨survivin与宫颈鳞癌新辅助化疗(NACT)和放射治疗(RT)敏感性的相关性.方法 新辅助化疗前后采用免疫组化(SABC法) 法,检测32例宫颈鳞癌组织中survivin的表达.结果 32例患者化疗前后survivin阳性表达率分别为68.8%(22/32)和56.3%(18/32),差异无统计学意义(P=0.302).<45岁与≥45岁、Ⅰb~Ⅱa期与Ⅱb~Ⅲb期、高中分化癌与低分化癌、化疗前与化疗后 survivin阳性表达率比较均无统计学意义(P>0.05).化疗前survivin阳性表达患者NACT与NACT+RT总体疗效比较,差异均有统计学意义(P=0.030,0.009);survivin(+)组和(++)组NACT与NACT+RT的疗效比较,差异均有统计学意义(P≤0.05);survivin(-)组和(+++)组NACT与NACT+RT的疗效比较,差异均无统计学意义(P≥0.05).结论 宫颈鳞癌survivin表达与患者年龄、临床分期和肿瘤分化程度无明显相关性,NACT和NACT+RT对不同survivin表达程度患者的疗效有显著性差异,治疗前检测患者survivin表达水平可为预测化疗和放疗敏感性提供客观依据.

  12. 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.

  13. 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

  14. 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

  15. Chimeric nucleolin aptamer with survivin DNAzyme for cancer cell targeted delivery.

    Science.gov (United States)

    Subramanian, Nithya; Kanwar, Jagat R; Akilandeswari, Balachandran; Kanwar, Rupinder K; Khetan, Vikas; Krishnakumar, Subramanian

    2015-04-25

    A chimeric aptamer-DNAzyme conjugate was generated for the first time using a nucleolin aptamer (NCL-APT) and survivin Dz (Sur_Dz) and exhibited the targeted killing of cancer cells. This proof of concept of using an aptamer for the delivery of DNAzyme can be applied to other cancer types to target survivin in cancer cells in a specific manner. PMID:25797393

  16. Tanshinone I induces human colorectal cancer cell apoptosis: The potential roles of Aurora A-p53 and survivin-mediated signaling pathways.

    Science.gov (United States)

    Lu, Mingjie; Wang, Chen; Wang, Jian

    2016-08-01

    Colorectal cancer (CRC) is one of the most common malignancies worldwide and a leading cause of cancer death. Despite decades of intensive investigations, effective interventional options are still limited and patient prognosis remains poor. Tanshinone I, an active compound from traditional Chinese herbal medicine Salvia miltiorrhiza Bunge, has been shown to inhibit cell growth of leukemia, lung, and breast cancers. However, whether and how Tanshinone I exerts similar effects on CRC needs to be elucidated. Tanshinone I induced CRC cell apoptosis was characterized and the roles of Aurora A-p53 and survivin-mediated pathways were analyzed in different CRC cell lines. Tanshinone I markedly inhibited CRC cell growth and induced apoptosis in CRC cells with functional p53 protein. Interestingly, Tanshinone I did not exert as much inhibitory effect on normal colon epithelial cells or CRC cells with mutant p53, indicating relative selectivity toward colorectal cancer cells with full presence of p53. In tse cells with wild-type p53, data showed that Tanshinone I mediated Aurora A inhibition results in p53 upregulation, which is required for cell apoptosis. In CRC cells with mutant p53 protein (not able to localize to the nucleus), however, Aurora A knockdown failed to induce CRC cell apoptosis. Instead, data showed that protein level of survivin decreased following Tanshinone I treatment. These observations were further substantiated by the pivotal role of survivin in Tanshinone I mediated apoptosis in CRC cells with p53 mutant. Tanshinone I, a novel natural compound, exerts significant inhibitory effect on CRC cell growth via a mechanism involving either Aurora A-p53 axis or survivin-involving mechanism depending on different intrinsic characteristics of tumor cells. PMID:27279458

  17. 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.

  18. 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.

  19. Expression of Bcl-2 in cells with different telomerase activities

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Both telomerase and Bcl-2 are important genes in controlling apoptosis. The activation of telomerase and the abnormal regulation of Bcl-2 are also closely related to carcinogenesis. However, little is known about the linkage between telomerase and Bcl-2. The effect of activated telomerase on the expression of Bcl-2 has been investigated. It is demonstrated that in tumor and transformed cells with higher telomerase activity, Bcl-2 expression is significantly lower than that in telomerase negative or less telomerose activity cells. Further study showed that in the telomerase gene-transformed 2BS-fibroblasts, Bcl-2 expression is inhibited significantly while the exogenous telomerase catalytic subunit gene is re-expressed in fibroblasts. Results indicated that there might be a certain linkage between the expression of telomerase and Bcl-2, and overexpression of exogenous telomerase gene might down regulate the expression of Bcl-2.

  20. 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.

  1. Identification and characterization of survivin-derived H-2Kb-restricted CTL epitopes

    DEFF Research Database (Denmark)

    Hofmann, Uta B; Voigt, Heike; Andersen, Mads H;

    2009-01-01

    for potential binding K(b)-restricted octamer peptide epitopes. Two epitopes, which bind strongly to K(b), were selected to test their immunogenicity in vivo. Spleen cells from mice vaccinated by intradermal injection of mature DC pulsed with these peptides displayed reactivity to the respective epitopes......Survivin is overexpressed in several malignancies and in tumor-associated endothelium making it an attractive target for therapeutic cytotoxic T-cell responses. Thus, it would be important to test this notion in preclinical models. Consequently, we screened the murine survivin sequence...... in a reduction of tumor cells but also the tumor supplying blood vessels. The presented preclinical model for survivin-directed vaccination may serve as a valuable tool to improve already running clinical trials in a syngeneic tumor model....

  2. Gene therapy for C-26 colon cancer using heparin-polyethyleneimine nanoparticle-mediated survivin T34A

    Directory of Open Access Journals (Sweden)

    Zhang L

    2011-10-01

    Full Text Available Ling Zhang1,*, Xiang Gao1,2,*, Ke Men1, BiLan Wang1, Shuang Zhang1, Jinfeng Qiu1, Meijuan Huang1, MaLing Gou1, Ning Huang2, ZhiYong Qian1, Xia Zhao1, YuQuan Wei11State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, 2Department of Pathophysiology, College of Preclinical and Forensic Medical Sciences, Sichuan University, Chengdu, People’s Republic of China*These authors contributed equally to this workBackground: Gene therapy provides a novel method for the prevention and treatment of cancer, but the clinical application of gene therapy is restricted, mainly because of the absence of an efficient and safe gene delivery system. Recently, we developed a novel nonviral gene carrier, ie, heparin-polyethyleneimine (HPEI nanoparticles for this purpose.Methods and results: HPEI nanoparticles were used to deliver plasmid-expressing mouse survivin-T34A (ms-T34A to treat C-26 carcinoma in vitro and in vivo. According to the in vitro studies, HPEI nanoparticles could efficiently transfect the pGFP report gene into C-26 cells, with a transfection efficiency of 30.5% ± 2%. Moreover, HPEI nanoparticle-mediated ms-T34A could efficiently inhibit the proliferation of C-26 cells by induction of apoptosis in vitro. Based on the in vivo studies, HPEI nanoparticles could transfect the Lac-Z report gene into C-26 cells in vivo. Intratumoral injection of HPEI nanoparticle-mediated ms-T34A significantly inhibited growth of subcutaneous C-26 carcinoma in vivo by induction of apoptosis and inhibition of angiogenesis.Conclusion: This research suggests that HPEI nanoparticle-mediated ms-T34A may have a promising role in C-26 colon carcinoma therapy.Keywords: gene therapy, mouse survivin-T34A, colon cancer, polyethyleneimine, nanoparticles, cancer therapy

  3. 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.

  4. 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

  5. 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...

  6. HPV detection and measurement of HPV-16, telomerase, and survivin transcripts in colposcopy clinic patients

    OpenAIRE

    Lanham, S; Herbert, A.; Watt, P

    2001-01-01

    Aims—To determine whether the detection of high risk human papillomavirus (HPV) types is more predictive for high grade CIN than the current cervical smear test, and whether the production and measurement of HPV type 16 (HPV-16) and cellular survivin and telomerase transcripts can be used to discriminate between cervical HPV infections that self cure and those that induce high grade lesions.

  7. p53 increases caspase-6 expression and activation in muscle tissue expressing mutant huntingtin

    DEFF Research Database (Denmark)

    Ehrnhoefer, Dagmar E; Skotte, Niels H; Ladha, Safia;

    2014-01-01

    a role in the peripheral phenotypes, such as muscle wasting observed in HD. We assessed skeletal muscle tissue from HD patients and well-characterized mouse models of HD. Cleavage of the caspase-6 specific substrate lamin A is significantly increased in skeletal muscle obtained from HD patients as well...... as in muscle tissues from two different HD mouse models. p53, a transcriptional activator of caspase-6, is upregulated in neuronal cells and tissues expressing mutant huntingtin. Activation of p53 leads to a dramatic increase in levels of caspase-6 mRNA, caspase-6 activity and cleavage of lamin A. Using mouse......-6 expression and activation is exacerbated in cells and tissues of both neuronal and peripheral origin expressing mutant huntingtin (Htt). These findings suggest that the presence of the mutant Htt protein enhances p53 activity and lowers the apoptotic threshold, which activates caspase-6...

  8. Learning Multiscale Active Facial Patches for Expression Analysis.

    Science.gov (United States)

    Zhong, Lin; Liu, Qingshan; Yang, Peng; Huang, Junzhou; Metaxas, Dimitris N

    2015-08-01

    In this paper, we present a new idea to analyze facial expression by exploring some common and specific information among different expressions. Inspired by the observation that only a few facial parts are active in expression disclosure (e.g., around mouth, eye), we try to discover the common and specific patches which are important to discriminate all the expressions and only a particular expression, respectively. A two-stage multitask sparse learning (MTSL) framework is proposed to efficiently locate those discriminative patches. In the first stage MTSL, expression recognition tasks are combined to located common patches. Each of the tasks aims to find dominant patches for each expression. Secondly, two related tasks, facial expression recognition and face verification tasks, are coupled to learn specific facial patches for individual expression. The two-stage patch learning is performed on patches sampled by multiscale strategy. Extensive experiments validate the existence and significance of common and specific patches. Utilizing these learned patches, we achieve superior performances on expression recognition compared to the state-of-the-arts. PMID:25291808

  9. Altered glutamyl-aminopeptidase activity and expression in renal neoplasms

    International Nuclear Information System (INIS)

    Advances in the knowledge of renal neoplasms have demonstrated the implication of several proteases in their genesis, growth and dissemination. Glutamyl-aminopeptidase (GAP) (EC. 3.4.11.7) is a zinc metallopeptidase with angiotensinase activity highly expressed in kidney tissues and its expression and activity have been associated wtih tumour development. In this prospective study, GAP spectrofluorometric activity and immunohistochemical expression were analysed in clear-cell (CCRCC), papillary (PRCC) and chromophobe (ChRCC) renal cell carcinomas, and in renal oncocytoma (RO). Data obtained in tumour tissue were compared with those from the surrounding uninvolved kidney tissue. In CCRCC, classic pathological parameters such as grade, stage and tumour size were stratified following GAP data and analyzed for 5-year survival. GAP activity in both the membrane-bound and soluble fractions was sharply decreased and its immunohistochemical expression showed mild staining in the four histological types of renal tumours. Soluble and membrane-bound GAP activities correlated with tumour grade and size in CCRCCs. This study suggests a role for GAP in the neoplastic development of renal tumours and provides additional data for considering the activity and expression of this enzyme of interest in the diagnosis and prognosis of renal neoplasms

  10. PUMA and survivin are involved in the apoptosis of HepG2 cells induced by microcystin-LR via mitochondria-mediated pathway.

    Science.gov (United States)

    Ma, Junguo; Feng, Yiyi; Liu, Yang; Li, Xiaoyu

    2016-08-01

    The present study aimed to determine the cytotoxicity of microcystin-LR (MC-LR) on the human hepatocellular carcinoma (HepG2) cells in order to elucidate the mechanism of apoptosis induced by MC-LR. Morphological evaluation results showed that MC-LR induced time- and concentration-dependent apoptosis in HepG2 cells. The biochemical assays revealed that MC-LR-exposure caused overproduction of reactive oxygen species (ROS), cyclooxygenase-2 activity alteration, cytochrome c release, and remarkable activation of caspase-3 and caspase-9 in HepG2 cells, indicating that MC-LR-induced apoptosis is mediated by mitochondrial pathway. Moreover, we also found that p53 and Bax might play an important role in MC-LR-induced apoptosis in HepG2 cells in which PUMA and survivin were involved. However, further studies are necessary to elucidate the possible functions of PUMA and survivin in MC-LR-induced apoptosis in HepG2 cells. PMID:27235693

  11. 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

  12. 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.

  13. 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.

  14. Cloning and Expression of Yak Active Chymosin in Pichia pastoris.

    Science.gov (United States)

    Luo, Fan; Jiang, Wei Hua; Yang, Yuan Xiao; Li, Jiang; Jiang, Ming Feng

    2016-09-01

    Rennet, a complex of enzymes found in the stomachs of ruminants, is an important component for cheese production. In our study, we described that yak chymosin gene recombinant Pichia pastoris strain could serve as a novel source for rennet production. Yaks total RNA was extracted from the abomasum of an unweaned yak. The yak preprochymosin, prochymosin, and chymosin genes from total RNA were isolated using gene specific primers based on cattle chymosin gene sequence respectively and analyzed their expression pattern byreal time-polymerase chain reaction. The result showed that the chymosin gene expression level of the sucking yaks was 11.45 times higher than one of adult yaks and yak chymosin belongs to Bovidae family in phylogenetic analysis. To express each, the preprochymosin, prochymosin, and chymosin genes were ligated into the expression vector pPICZαA, respectively, and were expressed in Pichia pastoris X33. The results showed that all the recombinant clones of P. pastoris containing the preprochymosin, prochymosin or chymosin genes could produce the active form of recombinant chymosin into the culture supernatant. Heterologous expressed prochymosin (14.55 Soxhlet unit/mL) had the highest enzyme activity of the three expressed chymosin enzymes. Therefore, we suggest that the yak chymosin gene recombinant Pichia pastoris strain could provide an alternative source of rennet production. PMID:27004812

  15. [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.

  16. 胰腺癌相关抗原MUC1与survivin mRNA联合转染树突细胞激发特异性细胞毒T淋巴细胞能力的体外研究%In vitro study of induction of specific cytotoxic T lymphocytes by the dendritic cells co-transfected with pancreatic cancer-associated antigen MUC1 and survivin mRNA

    Institute of Scientific and Technical Information of China (English)

    陈江; 郭晓钟; 李宏宇; 邵晓东; 王迪; 赵佳钧; 许文达

    2014-01-01

    分泌的IL-10的差异无统计学意义.结论 MUC1与survivin mRNA联合转染的DC较单一抗原转染的DC具有更强的体外特异性CTL激发能力.%Objective To investigate the ability of induction of specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cells (DCs) co-transfected with MUC1 and survivin mRNA of human pancreatic cancer,and to provide the experimental basis for the treatment of human pancreatic cancer with multi-epitope DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) of 6 patients with pancreatic cancer.Human pancreatic cancer cell line MiaPaCa-2 was routinely cultured,after being transcripted and amplified by RT-PCR,MUC1 and survivin mRNA were co-transfected or individually transfected into DCs by electroporation,and they were named as DC-MUC1,DC-survivin,DC-MUC1 + survivin.The expression of MUC1 and survivin mRNA in DCs were detected by real-time PCR.The survival rate of transfected DCs were determined by MTT method.The lymphocyte proliferation ability was evaluated by mixed cell culture method.The Th1 cytokine releasing of antigen-specific CTLs were measured by ELISA assay.Results Mature DCs were obtained,the positive expression rates of surface markers CD40,HLA-DR,CD83 and CD86 were 34.31%,50.21%,89.17% and 73.62%,respectively.The expression amount of MUC1 mRNA of DC-MUC1 was 36.24 ± 5.17,and the expression amount of survivin mRNA of DC-survivin was 34.53 ± 4.02,while the expression amounts of MUC1,survivin mRNA of DC-MUC1 + surviving were 31.79 ±4.26 and 14.67 ± 2.96,which were significantly lower than that in individual transfection group (P < 0.05).The survival rate of DC-MUC1 + surviving was decreased in a time dependent manner,which was significantly lower than that in individual transfection group (about 50.21% vs 80% at 24 h,P <0.05).When DC/T cells ratio was 1∶ 10,1∶ 20,the autologous T cell proliferation index of MUC1 and survivin mRNA in co-transfection DC

  17. Melatonin modulates aromatase activity and expression in endothelial cells.

    Science.gov (United States)

    Alvarez-García, Virginia; González, Alicia; Martínez-Campa, Carlos; Alonso-González, Carolina; Cos, Samuel

    2013-05-01

    Melatonin is known to suppress the development of endocrine-responsive breast cancers by interacting with the estrogen signaling pathways. Paracrine interactions between malignant epithelial cells and proximal stromal cells are responsible for local estrogen biosynthesis. In human breast cancer cells and peritumoral adipose tissue, melatonin downregulates aromatase, which transforms androgens into estrogens. The presence of aromatase on endothelial cells indicates that endothelial cells may contribute to tumor growth by producing estrogens. Since human umbilical vein endothelial cells (HUVECs) express both aromatase and melatonin receptors, the aim of the present study was to evaluate the ability of melatonin to regulate the activity and expression of aromatase on endothelial cells, thus, modulating local estrogen biosynthesis. In the present study, we demonstrated that melatonin inhibits the growth of HUVECs and reduces the local biosynthesis of estrogens through the downregulation of aromatase. These results are supported by three lines of evidence. Firstly, 1 mM of melatonin counteracted the testosterone-induced cell proliferation of HUVECs, which is dependent on the local biosynthesis of estrogens from testosterone by the aromatase activity of the cells. Secondly, we found that 1 mM of melatonin reduced the aromatase activity of HUVECs. Finally, by real‑time RT-PCR, we demonstrated that melatonin significantly downregulated the expression of aromatase as well as its endothelial-specific aromatase promoter region I.7. We conclude that melatonin inhibits aromatase activity and expression in HUVECs by regulating gene expression of specific aromatase promoter regions, thereby reducing the local production of estrogens. PMID:23450505

  18. Efficient inhibition of murine breast cancer growth and metastasis by gene transferred mouse survivin Thr34→Ala mutant

    Directory of Open Access Journals (Sweden)

    Chen li-Juan

    2008-09-01

    Full Text Available Abstract Background Metastasis in breast cancer is a vital concern in treatment because most women with primary breast cancer have micrometastases to distant sites at diagnosis. As a member of the inhibitor of apoptosis protein (IAP family, survivin has been proposed as an attractive target for new anticancer interventions. In this study, we investigated the role of the plasmid encoding the phosphorylation-defective mouse survivin threonine 34→alanine mutant (Msurvivin T34A plasmid in suppressing both murine primary breast carcinomas and pulmonary metastases. Methods In vitro study, induction of apoptosis by Msurvivin T34A plasmid complexed with cationic liposome (DOTAP/Chol was examined by PI staining fluorescence microscopy and flow cytometric analysis. The anti-tumor and anti-metastases activity of Msurvivin T34A plasmid complexed with cationic liposome (DOTAP/Chol was evaluated in female BALB/c mice bearing 4T1 s.c. tumors. Mice were treated twice weekly with i.v. administration of Msurvivin T34A plasmid complexed with cationic liposome (DOTAP/Chol, PORF-9 null plasmid complexed with cationic liposome (DOTAP/Chol, 0.9% NaCl solution for 4 weeks. Tumor volume was observed. After sacrificed, tumor net weight was measured and Lung metastatic nodules of each group were counted. Assessment of apoptotic cells by TUNEL assay was conducted in tumor tissue. Microvessel density within tumor tissue was determined by CD31 immunohistochemistry. Alginate-encapsulated tumor cells test was conducted to evaluate the effect on angiogenesis. By experiment of cytotoxicity T lymphocytes, we test whether Msurvivin T34A plasmid complexed with cationic liposome (DOTAP/Chol can induce specific cell immune response. Results Administration of Msurvivin T34A plasmid complexed with cationic liposome (DOTAP/Chol resulted in significant inhibition in the growth and metastases of 4T1 tumor model. These anti-tumor and anti-metastases responses were associated with

  19. 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.

  20. 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].

  1. 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.

  2. Expression and activation of proteases in co-cultures.

    Science.gov (United States)

    Paduch, Roman; Kandefer-Szerszeń, Martyna

    2011-01-01

    The present study concerned the expression and activation of metalloproteinase-2 (MMP-2), metalloproteinase-9 (MMP-9) and the urokinase plasminogen activator/urokinase plasminogen activator receptor (uPA/uPAR) system in co-cultures of human colon carcinoma cell spheroids (HT29, LS180, SW948) with human normal colon epithelium (CCD 841 CoTr), myofibroblasts (CCD-18Co) and endothelial cells (HUVEC). Additionally, the influence of monensin on the production and function of the proteases was tested. Tumor cells expressed small amounts of MMP-2, MMP-9 and uPA. Normal cells generally produced proportionally higher concentrations of these proteases (especially MMP-2, compared with significantly smaller yields of MMP-9 and significantly lower amounts of uPAR than tumors. In co-cultures of tumor spheroids with normal cell monolayers, the concentration of the proteases was equal to the sum of the enzymes produced in monocultures of both types of cells. The highest activity of uPA, measured as the reduction of the chromogenic substrate (S-2444), was detected in supernatants and lysates of endothelial cells. Interestingly, in normal cells, the higher expression of proteases, mainly uPA, measured as the level of protein concentration, was closely linked with their lower activity and inversely, in tumor cells, the low level of the expression of the enzymes correlated with their high enzymatic activity. In zymography analysis, mainly pro-MMPs were detected both in culture supernatants and cell lysates. The highest amounts of active forms of the MMPs were detected in tumor spheroids co-cultured with endothelial cells. Monensin inhibited MMPs and uPA secretion but significantly increased uPAR release, mainly from normal cells. In conclusion, during direct interactions of tumor cells with normal cells, MMPs and the uPA/uPAR system play an important role in the degradation of ECM and tumor development, but as we found, there is a reverse relationship between the concentration and the

  3. 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

  4. CRISPR-mediated Activation of Latent HIV-1 Expression.

    Science.gov (United States)

    Limsirichai, Prajit; Gaj, Thomas; Schaffer, David V

    2016-03-01

    Complete eradication of HIV-1 infection is impeded by the existence of cells that harbor chromosomally integrated but transcriptionally inactive provirus. These cells can persist for years without producing viral progeny, rendering them refractory to immune surveillance and antiretroviral therapy and providing a permanent reservoir for the stochastic reactivation and reseeding of HIV-1. Strategies for purging this latent reservoir are thus needed to eradicate infection. Here, we show that engineered transcriptional activation systems based on CRISPR/Cas9 can be harnessed to activate viral gene expression in cell line models of HIV-1 latency. We further demonstrate that complementing Cas9 activators with latency-reversing compounds can enhance latent HIV-1 transcription and that epigenome modulation using CRISPR-based acetyltransferases can also promote viral gene activation. Collectively, these results demonstrate that CRISPR systems are potentially effective tools for inducing latent HIV-1 expression and that their use, in combination with antiretroviral therapy, could lead to improved therapies for HIV-1 infection. PMID:26607397

  5. Equine 5α-reductase activity and expression in epididymis.

    Science.gov (United States)

    Corbin, C J; Legacki, E L; Ball, B A; Scoggin, K E; Stanley, S D; Conley, A J

    2016-10-01

    The 5α-reductase enzymes play an important role during male sexual differentiation, and in pregnant females, especially equine species where maintenance relies on 5α-reduced progesterone, 5α-dihydroprogesterone (DHP). Epididymis expresses 5α-reductases but was not studied elaborately in horses. Epididymis from younger and older postpubertal stallions was divided into caput, corpus and cauda and examined for 5α-reductase activity and expression of type 1 and 2 isoforms by quantitative real-time polymerase chain reaction (qPCR). Metabolism of progesterone and testosterone to DHP and dihydrotestosterone (DHT), respectively, by epididymal microsomal protein was examined by thin-layer chromatography and verified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Relative inhibitory potencies of finasteride and dutasteride toward equine 5α-reductase activity were investigated. Pregnenolone was investigated as an additional potential substrate for 5α-reductase, suggested previously from in vivo studies in mares but never directly examined. No regional gradient of 5α-reductase expression was observed by either enzyme activity or transcript analysis. Results of PCR experiments suggested that type 1 isoform predominates in equine epididymis. Primers for the type 2 isoform were unable to amplify product from any samples examined. Progesterone and testosterone were readily reduced to DHP and DHT, and activity was effectively inhibited by both inhibitors. Using epididymis as an enzyme source, no experimental evidence was obtained supporting the notion that pregnenolone could be directly metabolized by equine 5α-reductases as has been suggested by previous investigators speculating on alternative metabolic pathways leading to DHP synthesis in placenta during equine pregnancies. PMID:27466384

  6. Re-expression of HPV16 E2 in SiHa (human cervical cancer) cells potentiates NF-κB activation induced by TNF-α concurrently increasing senescence and survival.

    Science.gov (United States)

    Prabhavathy, Devan; Subramanian, Chandrasekaran Karthik; Karunagaran, Devarajan

    2015-01-01

    Re-expression of E2 in human papillomavirus (HPV) transformed tumour cells can induce apoptosis; however, some evidences also attribute an important role to E2 in sustaining tumorigenesis. In the present paper, we studied the effects of tumour necrosis factor (TNF)-α-mediated NF-κB (nuclear factor kappa-light-chain-enhancer of activated B-cells) activation on E2-induced senescence in HPV16-integrated SiHa cells. The results show that E2 inhibits endogenous E6 gene expression and sensitizes SiHa cells to TNF-α-induced NF-κB activation. Under this condition there was an increase in the expression of senescent proteins p53, p21, p27 and p16 and senescence-associated (SA)-β-galactosidase activity indicating that TNF-α augments E2-mediated senescence. Re-expression of E2 expression with TNF-α treatment resulted in an increase in the expression of anti-apoptotic Bcl2 (B-cell lymphoma 2) protein and other pro-survival genes like cyclin D1 (cyc D1), survivin and hTERT (human telomerase reverse transcriptase). Concomitantly, E2 + TNF-α combination increased the survival of SiHa cells by positive changes in viability, proliferation and colony formation. E2-induced apoptotic tendency shifted towards senescence in presence of TNF-α by arresting the cells at both G0/G1 and G2/M phases, thus enhancing cell survival. Another observation in the present study is the significant up-regulation of key senescence messaging factors regulated by NF-κB namely interleukin (IL)-6, IL-8, high-mobility group protein A (HMGA)1 and B (HMGB)1 in E2-transfected cells treated with TNF-α. Our data provide a mechanistic basis and a new insight for the role of TNF-α and E2 in linking cellular senescence, tumorigenesis and HPV re-infection. PMID:25572145

  7. 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

  8. Behavioral meaningful opioidergic stimulation activates kappa receptor gene expression

    International Nuclear Information System (INIS)

    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

  9. Neurotrophin receptors expression and JNK pathway activation in human astrocytomas

    International Nuclear Information System (INIS)

    Neurotrophins are growth factors that regulate cell growth, differentiation and apoptosis in the nervous system. Their diverse actions are mediated through two different transmembrane – receptor signaling systems: Trk receptor tyrosine kinases (TrkA, TrkB, TrkC) and p75NTR neurotrophin receptor. Trk receptors promote cell survival and differentiation while p75NTR induces, in most cases, the activity of JNK-p53-Bax apoptosis pathway or suppresses intracellular survival signaling cascades. Robust Trk activation blocks p75NTR -induced apoptosis by suppressing the JNK-p53-Bax pathway. The aim of this exploratory study was to investigate the expression levels of neurotrophin receptors, Trks and p75NTR, and the activation of JNK pathway in human astrocytomas and in adjacent non-neoplastic brain tissue. Formalin-fixed paraffin-embedded serial sections from 33 supratentorial astrocytomas (5 diffuse fibrillary astrocytomas, WHO grade II; 6 anaplastic astrocytomas, WHO grade III; 22 glioblastomas multiforme, WHO grade IV) were immunostained following microwave pretreatment. Polyclonal antibodies against TrkA, TrkB, TrkC and monoclonal antibodies against p75NTR and phosphorylated forms of JNK (pJNK) and c-Jun (pc-Jun) were used. The labeling index (LI), defined as the percentage of positive (labeled) cells out of the total number of tumor cells counted, was determined. Moderate to strong, granular cytoplasmic immunoreactivity for TrkA, TrkB and TrkC receptors was detected in greater than or equal to 10% of tumor cells in the majority of tumors independently of grade; on the contrary, p75NTR receptor expression was found in a small percentage of tumor cells (~1%) in some tumors. The endothelium of tumor capillaries showed conspicuous immunoreactivity for TrkB receptor. Trk immunoreactivity seemed to be localized in some neurons and astrocytes in non-neoplastic tissue. Phosphorylated forms of JNK (pJNK) and c-Jun (pc-Jun) were significantly co-expressed in a tumor grade

  10. 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

  11. 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.

  12. Mitotic arrest and apoptosis in breast cancer cells induced by Origanum majorana extract: upregulation of TNF-α and downregulation of survivin and mutant p53.

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    Yusra Al Dhaheri

    Full Text Available BACKGROUND: In the present study, we investigated the effect of Origanum majorana ethanolic extract on the survival of the highly proliferative and invasive triple-negative p53 mutant breast cancer cell line MDA-MB-231. RESULTS: We found that O. majorana extract (OME was able to inhibit the viability of the MDA-MB-231 cells in a time- and concentration-dependent manner. The effect of OME on cellular viability was further confirmed by the inhibition of colony growth. We showed, depending on the concentration used, that OME elicited different effects on the MDA-MB 231 cells. Concentrations of 150 and 300 µg/mL induced an accumulation of apoptotic-resistant population of cells arrested in mitotis and overexpressing the cyclin-dependent kinase inhibitor, p21 and the inhibitor of apoptosis, survivin. On the other hand, higher concentrations of OME (450 and 600 µg/mL triggered a massive apoptosis through the extrinsic pathway, including the activation of tumor necrosis factor-α (TNF-α, caspase 8, caspase 3, and cleavage of PARP, downregulation of survivin as well as depletion of the mutant p53 in MDA-MB-231 cells. Furthermore, OME induced an upregulation of γ-H2AX, a marker of double strand DNA breaks and an overall histone H3 and H4 hyperacetylation. CONCLUSION: Our findings provide strong evidence that O. majorana may be a promising chemopreventive and therapeutic candidate against cancer especially for highly invasive triple negative p53 mutant breast cancer; thus validating its complementary and alternative medicinal use.

  13. 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.

  14. 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

  15. TLR3 expression correlates with apoptosis, proliferation and angiogenesis in hepatocellular carcinoma and predicts prognosis

    International Nuclear Information System (INIS)

    Toll-like receptor 3 (TLR3) plays a key role in innate immunity. In the present study, we analyzed tissues of patients with human hepatocellular carcinoma (HCC) to determine the significance of the relationship between TLR3 expression and cell proliferation, apoptosis, hepatitis B virus infections, angiogenesis and prognosis. We collected paraffin-embedded tissues from 85 patients with HCC who had complete histories and were followed for >5 years. The expression and intracellular localization of TLR3 and downstream proteins (TRIF, NF-κB, and IRF3) were detected using immunohistochemistry. Further, we determined the expression of proteins that mediate cell proliferation (Ki67, cyclin D1), apoptosis (survivin, bcl-2, caspases 3, 8, and 9), and angiogenesis (CD34, MMP-2) as well as the HBV proteins HBsAg and HBcAg. Apoptosis in HCC tissues was detected using TUNEL. We conducted dual-labeling immunohistochemical analyses of TLR3 expression and TUNEL activity. TLR3 expression was significantly lower in HCC tissues compared with adjacent tissues. TRIF, NF-κB, and IRF3 correlated positively with TLR3 expression. Survivin and Bcl-2 expression correlated negatively with TLR3. The frequencies of caspases 3, 8, and 9 expression correlated positively with TLR3 signaling proteins. Cytoplasmic TLR3 and serum levels of HBsAg correlated positively. The apoptotic index determined using the TUNEL method and correlated positively with TLR3 expression. TLR3 expression in the cytoplasm correlated positively with TUNEL-positive cells and HBsAg. Ki67 and cyclin D1 correlated negatively with TLR3 expression. MMP-2 expression, microvessel density (CD34+) and endothelial progenitor cells (EPCs) correlated negatively with TLR3 expression. Kaplan–Meier survival analysis shows that TLR3 expression correlated with longer survival. The expression of TLR3 in HCC tissues may exert a synergistic effect on apoptosis and inhibit the proliferation of HCC cells, MMP-2 expression, generation of EPCs

  16. 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.

  17. Troglitazone, a peroxisome proliferator-activated receptor γ ligand, induces growth inhibition and apoptosis of HepG2 human liver cancer cells

    Institute of Scientific and Technical Information of China (English)

    Yan-Ming Zhou; Yin-Hao Wen; Xiao-Yan Kang; Hai-Hua Qian; Jia-Mei Yang; Zheng-Feng Yin

    2008-01-01

    AIM: To examine the effect of troglitazone, a peroxisome proliferator-activated receptor γ (PPARγ) ligand, on the proliferation and apoptosis of human liver cancer cells.METHODS: Liver cancer cell line HepG2 was cultured and treated with troglitazone. Cell proliferation was detected by 3-(4-,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; apoptosis was detected by flow cytometry and terminal deoxynucleotidyl transferasemediated nick end labeling of DNA fragmentation sites (TUNEL) assay; and apoptosis-related protein was detected by immunocytochemistry and Western blotting.RESULTS: Troglitazone inhibited growth and induced apoptosis of HepG2 cells in a dose-dependent manner,and induced activation of caspase-3 expression.Troglitazone not only drove apoptosis-inhibiting factor survivin to translocate incompletely from the nucleus to the cytoplasm, but also inhibited expression of survivin,while it did not affect expression of apoptosis-promoting factor Bax.CONCLUSION: PPARγ ligands inhibit growth and induce apoptosis of liver cancer cells, and may have applications for the prevention and treatment of liver cancer.

  18. Pro-oncogene Pokemon promotes breast cancer progression by upregulating survivin expression

    OpenAIRE

    Zu, Xuyu; Ma, Jun; Liu, Hongxia; Liu, Feng; Tan, Chunyan; Yu, Lingling; Wang, Jue; Xie, Zhenhua; Cao, Deliang; Jiang, Yuyang

    2011-01-01

    Introduction Pokemon is an oncogenic transcription factor involved in cell growth, differentiation and oncogenesis, but little is known about its role in human breast cancer. In this study, we aimed to reveal the role of Pokemon in breast cancer progression and patient survival and to understand its underlying mechanisms. Methods Tissue microarray analysis of breast cancer tissues from patients with complete clinicopathological data and more than 20 years of follow-up were used to evaluate Po...

  19. 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.

  20. Combination of survivin siRNA with neoadjuvant chemotherapy enhances apoptosis and reverses drug resistance in breast cancer MCF-7 cells

    Directory of Open Access Journals (Sweden)

    Honglin Dong

    2015-01-01

    Conclusion: Survivin siRNA combined with the neoadjuvant chemotherapy can significantly enhance the sensitivity of MCF-7 cells to chemotherapeutics and cell apoptosis. This technology has important potential value in the therapeutic study of breast cancer.

  1. EGF activates TTP expression by activation of ELK-1 and EGR-1 transcription factors

    Directory of Open Access Journals (Sweden)

    Florkowska Magdalena

    2012-03-01

    Full Text Available Abstract Background Tristetraprolin (TTP is a key mediator of processes such as inflammation resolution, the inhibition of autoimmunity and in cancer. It carries out this role by the binding and degradation of mRNA transcripts, thereby decreasing their half-life. Transcripts modulated by TTP encode proteins such as cytokines, pro-inflammatory agents and immediate-early response proteins. TTP can also modulate neoplastic phenotypes in many cancers. TTP is induced and functionally regulated by a spectrum of both pro- and anti-inflammatory cytokines, mitogens and drugs in a MAPK-dependent manner. So far the contribution of p38 MAPK to the regulation of TTP expression and function has been best described. Results Our results demonstrate the induction of the gene coding TTP (ZFP36 by EGF through the ERK1/2-dependent pathway and implicates the transcription factor ELK-1 in this process. We show that ELK-1 regulates ZFP36 expression by two mechanisms: by binding the ZFP36 promoter directly through ETS-binding site (+ 883 to +905 bp and by inducing expression of EGR-1, which in turn increases ZFP36 expression through sequences located between -111 and -103 bp. Conclusions EGF activates TTP expression via ELK-1 and EGR-1 transcription factors.

  2. Activity and expression of urokinase-type plasminogen activator and matrix metalloproteinases in human colorectal cancer

    International Nuclear Information System (INIS)

    Matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and urokinase-type plasminogen activator (uPA) are involved in colorectal cancer invasion and metastasis. There is still debate whether the activity of MMP-2 and MMP-9 differs between tumors located in the colon and rectum. We designed this study to determine any differences in the expression of MMP-2, MMP-9 and uPA system between colon and rectal cancer tissues. Cancer tissue samples were obtained from colon carcinoma (n = 12) and rectal carcinomas (n = 10). MMP-2 and MMP-9 levels were examined using gelatin zymography and Western blotting; their endogenous inhibitors, tissue inhibitor of metalloproteinase-2 (TIMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1), were assessed by Western blotting. uPA, uPAR and PAI-1 were examined using enzyme-linked immunosorbent assay (ELISA). The activity of uPA was assessed by casein-plasminogen zymography. In both colon and rectal tumors, MMP-2, MMP-9 and TIMP-1 protein levels were higher than in corresponding paired normal mucosa, while TIMP-2 level in tumors was significantly lower than in normal mucosa. The enzyme activities or protein levels of MMP-2, MMP-9 and their endogenous inhibitors did not reach a statistically significant difference between colon and rectal cancer compared with their normal mucosa. In rectal tumors, there was an increased activity of uPA compared with the activity in colon tumors (P = 0.0266), however urokinase-type plasminogen activator receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1) showed no significant difference between colon and rectal cancer tissues. These findings suggest that uPA may be expressed differentially in colon and rectal cancers, however, the activities or protein levels of MMP-2, MMP-9, TIMP-1, TIMP-2, PAI-1 and uPAR are not affected by tumor location in the colon or the rectum

  3. 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.

  4. Correlation between expression of human telomerase subunits and telomerase activity in esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    Chun Li; Ming-Yao Wu; Ying-Rui Liang; Xian-Ying Wu

    2003-01-01

    AIM: To investigate telomerase activity and hTERT, TP-1 expression and their relationships in esophageal squamouscell carcinoma (ESCC).METHODS: Telomerase activity was measured in 60 ESCCtissues using telomeric repeat amplification protocol (TRAP)assay by silver staining. In situ hybridization was used for detecting hTERT and TP-lmRNA.RESULTS: The telomerase activity was detected in 83.3 % of ESCC tissues. The difference of telomerase activity was significant between well and poorly cancer differentiated lesions (P<0.05). The positive rate of telomerase activity was higher in patients with lymphatic metastasis than in patients without lymphatic metastasis. In cancer tissues hTERT mRNA expression was 75 % and TP-1 mRNA expression was 71.7 %. The expression of hTERT, TP-1 mRNA in well and poorly differentiated carcinoma was not significant. The expression of hTERT mRNA was correlated with telomerase activity, but TP-1 mRNA expression was not correlated with it.CONCLUSION: Telomerase activity and hTERT, TP-1 mRNA expression are up-regulated in ESCC. Telomerase activity in ESCC is correlated with lymphatic metastasis and cancer differentiation. Telomerase activity may be used as a prognostic marker in ESCC. hTERT mRNA expression is correlated with telomerase activity. Enhanced hTERT mRNA expression may initially comprehend the telomerase activity level, but it is less sensitive than TRAP assay.

  5. The heat shock protein 90 inhibitor SNX5422 has a synergistic activity with histone deacetylase inhibitors in induction of death of anaplastic thyroid carcinoma cells.

    Science.gov (United States)

    Kim, Si Hyoung; Kang, Jun Goo; Kim, Chul Sik; Ihm, Sung-Hee; Choi, Moon Gi; Yoo, Hyung Joon; Lee, Seong Jin

    2016-02-01

    The influence of the heat shock protein 90 (hsp90) inhibitor SNX5422 alone or in combination with the histone deacetylase (HDAC) inhibitors PXD101, suberoylanilide hydroxamic acid (SAHA), and trichostatin A (TSA) on survival of anaplastic thyroid carcinoma (ATC) cells was investigated. In 8505C and CAL62 cells, SNX5422 caused cell death with concomitant changes in the expression of hsp90 client proteins. After treatment of both SNX5422 and PXD101, SAHA and TSA, compared with treatment of SNX5422 alone, cell viability was diminished, whereas inhibition rate and cytotoxic activity were enhanced. All of the combination index values were lower than 1.0, suggesting the synergism between SNX5422 and PXD101, SAHA and TSA in induction of cell death. In cells treated with both SNX5422 and PXD101, SAHA and TSA, compared with cells treated with SNX5422 alone, the protein levels of Akt, phospho-4EBP1, phospho-S6 K, and survivin were diminished, while those of γH2AX, acetyl. histone H3, acetyl. histone H4, cleaved PARP, and cleaved caspase-3 were enhanced. In conclusion, these results demonstrate that SNX5422 has a cytotoxic activity in conjunction with alterations in the expression of hsp90 client proteins in ATC cells. Moreover, SNX5422 synergizes with HDAC inhibitors in induction of cytotoxicity accompanied by the suppression of PI3K/Akt/mTOR signaling and survivin, and the overexpression of DNA damage-related proteins in ATC cells. PMID:26219406

  6. Survivin基因沉默抑制胃癌MGC-803细胞增殖并增强其对塞来昔布的敏感性%Survivin gene silencing inhibits the proliferation of human gastric cancer MGC-803 cells and enhances their sensitivity to celecoxib

    Institute of Scientific and Technical Information of China (English)

    方雷; 吴海滨; 陈晓岗

    2011-01-01

    Objective To investigate the effect of small interfering RNA (siRNA)-mediated survivin knock-down on the proliferation of human gastric cancer MGC-803 cells and their sensitivity to celecoxib. Methods The siRNA against survivin was constructed and transfected into MGC-803 cells via LipofectamineTM 2000. The expression of survivin in the transfected cells was detected by RT-PCR and Western blot, and flow cytometry was used to detect the cell cycle changes. The sensitivity of the cells to celecoxib after transfection was examined using MTT assay and clonogenic assay. Results The protein and mRNA levels of survivin in MGC-803 cells were decreased significantly after siRNA transfection, which also caused cell cycle arrest in G0/G1, phase. The sensitivity of MGC-803 cells to celecoxib was significantly increased after siRNA transfection. Conclusion siRNA-mediated survivin silencing causes growth suppression of MGC-803 cells and enhances their sensitivity to celecoxib in vitro.%目的 研究Survivin基因沉默对人胃癌MGC-803细胞增殖和对化疗药物塞来昔布敏感性的影响.方法 设计合成Survivin的siRNA序列,LipofectamineTM2000转染入MGC-803细胞.采用RT-PCR和Western blotting检测Survivin在干扰后mRNA和蛋白的表达情况,利用流式细胞仪检测细胞周期.通过MTT法和细胞克隆形成试验法察Survivin基因沉默后MGC-803细胞对塞来昔布的敏感性.结果 Survivin基因沉默48 h后,MGC-803细胞的Survivin基因和蛋白表达明显降低(P<0.05).细胞周期被阻滞在G0/G1期,S期细胞数减少(P<0.05).Survivin基因沉默组细胞对塞来昔布的敏感性显著性增强(P<0.05).结论 Survivin特异性siRNA能显著沉默MGC-803细胞Survivin基因,抑制细胞增殖,并增强MGC-803细胞对塞来昔布的敏感性.

  7. 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.

  8. 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.

  9. 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...

  10. 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. PMID:26507667

  11. Inhibitors of apoptosis proteins (IAPs) expression and their prognostic significance in hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Similarly to other tumor types, an imbalance between unrestrained cell proliferation and impaired apoptosis appears to be a major unfavorable feature of hepatocellular carcinoma (HCC). The members of IAP family are key regulators of apoptosis, cytokinesis and signal transduction. IAP survival action is antagonized by specific binding of Smac/DIABLO and XAF1. This study aimed to investigate the gene and protein expression pattern of IAP family members and their antagonists in a series of human HCCs and to assess their clinical significance. Relative quantification of IAPs and their antagonist genes was assessed by quantitative Real Time RT-PCR (qPCR) in 80 patients who underwent surgical resection for HCC. The expression ratios of XIAP/XAF1 and of XIAP/Smac were also evaluated. Survivin, XIAP and XAF1 protein expression were investigated by immunohistochemistry. Correlations between mRNA levels, protein expression and clinicopathological features were assessed. Follow-up data were available for 69 HCC patients. The overall survival analysis was estimated according to the Kaplan-Meier method. Survivin and Livin/ML-IAP mRNAs were significantly over-expressed in cancer tissues compared to non-neoplastic counterparts. Although Survivin immunoreactivity did not correlate with qPCR data, a significant relation was found between higher Survivin mRNA level and tumor stage, tumor grade and vascular invasion. The mRNA ratio XIAP/XAF1 was significantly higher in HCCs than in cirrhotic tissues. Moreover, high XIAP/XAF1 ratio was an indicator of poor prognosis when overall survival was estimated and elevated XIAP immunoreactivity was significantly associated with shorter survival. Our study demonstrates that alterations in the expression of IAP family members, including Survivin and Livin/ML-IAP, are frequent in HCCs. Of interest, we could determine that an imbalance in XIAP/XAF1 mRNA expression levels correlated to overall patient survival, and that high XIAP immunoreactivity

  12. Expression of functionally active sialylated human erythropoietin in plants

    OpenAIRE

    Jez, Jakub; Castilho, Alexandra; Grass, Josephine; Vorauer-Uhl, Karola; Sterovsky, Thomas; Altmann, Friedrich; Steinkellner, Herta

    2013-01-01

    Recombinant human erythropoietin (rhEPO), a glycohormone, is one of the leading biopharmaceutical products. The production of rhEPO is currently restricted to mammalian cell expression systems because of rhEPO's highly complex glycosylation pattern, which is a major determinant for drug-efficacy. Here we evaluate the ability of plants to produce different glycoforms of rhEPO. cDNA constructs were delivered to Nicotiana benthamiana (N. benthamiana) and transiently expressed by a viral based ex...

  13. Expression and assembly of a fully active antibody in algae

    OpenAIRE

    Mayfield, Stephen P.; Franklin, Scott E.; Lerner, Richard A.

    2003-01-01

    Although combinatorial antibody libraries have solved the problem of access to large immunological repertoires, efficient production of these complex molecules remains a problem. Here we demonstrate the efficient expression of a unique large single-chain (lsc) antibody in the chloroplast of the unicellular, green alga, Chlamydomonas reinhardtii. We achieved high levels of protein accumulation by synthesizing the lsc gene in chloroplast codon bias and by driving expression of the chimeric gene...

  14. 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.

  15. 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.

  16. Activation of Peroxisome Proliferator-activated Receptor γ (PPARγ) and CD36 Protein Expression: THE DUAL PATHOPHYSIOLOGICAL ROLES OF PROGESTERONE.

    Science.gov (United States)

    Yang, Xiaoxiao; Zhang, Wenwen; Chen, Yuanli; Li, Yan; Sun, Lei; Liu, Ying; Liu, Mengyang; Yu, Miao; Li, Xiaoju; Han, Jihong; Duan, Yajun

    2016-07-15

    Progesterone or its analog, one of components of hormone replacement therapy, may attenuate the cardioprotective effects of estrogen. However, the underlying mechanisms have not been fully elucidated. Expression of CD36, a receptor for oxidized LDL (oxLDL) that enhances macrophage/foam cell formation, is activated by the transcription factor peroxisome proliferator-activated receptor γ (PPARγ). CD36 also functions as a fatty acid transporter to influence fatty acid metabolism and the pathophysiological status of several diseases. In this study, we determined that progesterone induced macrophage CD36 expression, which is related to progesterone receptor (PR) activity. Progesterone enhanced cellular oxLDL uptake in a CD36-dependent manner. Mechanistically, progesterone increased PPARγ expression and PPARγ promoter activity in a PR-dependent manner and the binding of PR with the progesterone response element in the PPARγ promoter. Specific deletion of macrophage PPARγ (MφPPARγ KO) expression in mice abolished progesterone-induced macrophage CD36 expression and cellular oxLDL accumulation. We also determined that, associated with gestation and increased serum progesterone levels, CD36 and PPARγ expression in mouse adipose tissue, skeletal muscle, and peritoneal macrophages were substantially activated. Taken together, our study demonstrates that progesterone can play dual pathophysiological roles by activating PPARγ expression, in which progesterone increases macrophage CD36 expression and oxLDL accumulation, a negative effect on atherosclerosis, and enhances the PPARγ-CD36 pathway in adipose tissue and skeletal muscle, a protective effect on pregnancy. PMID:27226602

  17. Expression of Activated Epidermal Growth Factor Receptor and Transcription Factor E2F in Condyloma Accuminata

    Institute of Scientific and Technical Information of China (English)

    俞小虹; 程浩; 郑伟

    2003-01-01

    Objective: To study the expression of activated epi-dermal growth factor receptor (EGFR) and transcrip-tion factor E2F (E2F) in Condyloma Accuminata(CA) patients. Methods: Immunofluorescent techniques were used to investigate the expression of activated EGFR and E2F in CA patients. Results: The expression of activated EGFR on the membrane of epithelial cells in CA lesions was sig-nificantly greater compared to expression levers in the control group (P<0.01). Moreover, the co-expres-sion of activated EGFR and E2F was significantly in-creased compared to the control group (P<0.01).Conclusion: Our observations suggest that the in-crease in activated EGFR expression may stimulate hyperplasia in CA patients through the activation of transcription factor E2F.

  18. 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.

  19. 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.

  20. Expression and assembly of a fully active antibody in algae

    Science.gov (United States)

    Mayfield, Stephen P.; Franklin, Scott E.; Lerner, Richard A.

    2003-01-01

    Although combinatorial antibody libraries have solved the problem of access to large immunological repertoires, efficient production of these complex molecules remains a problem. Here we demonstrate the efficient expression of a unique large single-chain (lsc) antibody in the chloroplast of the unicellular, green alga, Chlamydomonas reinhardtii. We achieved high levels of protein accumulation by synthesizing the lsc gene in chloroplast codon bias and by driving expression of the chimeric gene using either of two C. reinhardtii chloroplast promoters and 5' and 3' RNA elements. This lsc antibody, directed against glycoprotein D of the herpes simplex virus, is produced in a soluble form by the alga and assembles into higher order complexes in vivo. Aside from dimerization by disulfide bond formation, the antibody undergoes no detectable posttranslational modification. We further demonstrate that accumulation of the antibody can be modulated by the specific growth regime used to culture the alga, and by the choice of 5' and 3' elements used to drive expression of the antibody gene. These results demonstrate the utility of alga as an expression platform for recombinant proteins, and describe a new type of single chain antibody containing the entire heavy chain protein, including the Fc domain.

  1. The aryl hydrocarbon receptor-dependent disruption of contact inhibition in rat liver WB-F344 epithelial cells is linked with induction of survivin, but not with inhibition of apoptosis

    International Nuclear Information System (INIS)

    Inhibition of apoptosis by the ligands of the aryl hydrocarbon receptor (AhR) has been proposed to play a role in their tumor promoting effects on liver parenchymal cells. However, little is presently known about the impact of toxic AhR ligands, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on apoptosis in other liver cell types, such as in liver epithelial/progenitor cells. In the present study, we focused on the effects of TCDD on apoptosis regulation in a model of liver progenitor cells, rat WB-F344 cell line, during the TCDD-elicited release from contact inhibition. The stimulation of cell proliferation in this cell line was associated with deregulated expression of a number of genes known to be under transcriptional control of the Hippo signaling pathway, a principal regulatory pathway involved in contact inhibition of cell proliferation. Interestingly, we found that mRNA and protein levels of survivin, a known Hippo target, which plays a role both in cell division and inhibition of apoptosis, were significantly up-regulated in rat liver epithelial cell model, as well as in undifferentiated human liver HepaRG cells. Using the short interfering RNA-mediated knockdown, we confirmed that survivin plays a central role in cell division of WB-F344 cells. When evaluating the effects of TCDD on apoptosis induction by camptothecin, a genotoxic topoisomerase I inhibitor, we observed that the pre-treatment of WB-F344 cells with TCDD increased number of cells with apoptotic nuclear morphology, and it potentiated cleavage of both caspase-3 and poly(ADP-ribose) polymerase I. This indicated that despite the observed up-regulation of survivin, apoptosis induced by the genotoxin was potentiated in the model of rat liver progenitor cells. The present results indicate that, unlike in hepatocytes, AhR agonists may not prevent induction of apoptosis elicited by DNA-damaging agents in a model of rat liver progenitor cells

  2. 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.

  3. Constitutive Expression of Functionally Active Protease-Activated Receptors 1 and 2 in Human Conjunctival Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Timothy J. Nickel

    2006-01-01

    epithelial cells as in vitro model. We performed RT-PCR and immunofluorescence analyses to determine the expression of PAR1 and PAR2, and monitored the production of IL-6 after activating HCECs with PAR1 activating agents (thrombin or TFLLRN or PAR2 activating agents (tryptase, trypsin, or SLIGKV. The results show that HCECs constitutively express PAR1 and PAR2 mRNA and proteins, and produce significant amounts of IL-6 when incubated with specific PAR-activating enzymes or agonist peptides. Thrombin- and tryptase-induced HCEC activation was blocked by PAR1 and PAR2 neutralizing antibodies, respectively, and by specific enzyme inhibitors. The constitutive expression of PAR1 and PAR2, and their activation by thrombin and tryptase, respectively, may have important implications in ocular inflammation.

  4. 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.

  5. Gene expression analyses implicate an alternative splicing program in regulating contractile gene expression and serum response factor activity in mice.

    Directory of Open Access Journals (Sweden)

    Twishasri Dasgupta

    Full Text Available Members of the CUG-BP, Elav-like family (CELF regulate alternative splicing in the heart. In MHC-CELFΔ transgenic mice, CELF splicing activity is inhibited postnatally in heart muscle via expression of a nuclear dominant negative CELF protein under an α-myosin heavy chain promoter. MHC-CELFΔ mice develop dilated cardiomyopathy characterized by alternative splicing defects, enlarged hearts, and severe contractile dysfunction. In this study, gene expression profiles in the hearts of wild type, high- and low-expressing lines of MHC-CELFΔ mice were compared using microarrays. Gene ontology and pathway analyses identified contraction and calcium signaling as the most affected processes. Network analysis revealed that the serum response factor (SRF network is highly affected. Downstream targets of SRF were up-regulated in MHC-CELFΔ mice compared to the wild type, suggesting an increase in SRF activity. Although SRF levels remained unchanged, known inhibitors of SRF activity were down-regulated. Conversely, we found that these inhibitors are up-regulated and downstream SRF targets are down-regulated in the hearts of MCKCUG-BP1 mice, which mildly over-express CELF1 in heart and skeletal muscle. This suggests that changes in SRF activity are a consequence of changes in CELF-mediated regulation rather than a secondary result of compensatory pathways in heart failure. In MHC-CELFΔ males, where the phenotype is only partially penetrant, both alternative splicing changes and down-regulation of inhibitors of SRF correlate with the development of cardiomyopathy. Together, these results strongly support a role for CELF-mediated alternative splicing in the regulation of contractile gene expression, achieved in part through modulating the activity of SRF, a key cardiac transcription factor.

  6. Activity-dependent brain-derived neurotrophic factor expression regulates cortistatin-interneurons and sleep behavior

    Directory of Open Access Journals (Sweden)

    Martinowich Keri

    2011-03-01

    Full Text Available Abstract Background Sleep homeostasis is characterized by a positive correlation between sleep length and intensity with the duration of the prior waking period. A causal role for brain-derived neurotrophic factor (BDNF in sleep homeostasis has been suggested, but the underlying mechanisms remain unclear. Cortistatin, a neuropeptide expressed primarily in a subset of cortical GABAergic interneurons, is another molecule implicated in sleep homeostasis. Results We confirmed that sleep deprivation leads to an increase in cortical cortistatin mRNA expression. Disruption of activity-dependent BDNF expression in a genetically modified mouse line impairs both baseline levels of cortistatin mRNA as well as its levels following sleep deprivation. Disruption of activity-dependent BDNF also leads to a decrease in sleep time during the active (dark phase. Conclusion Our studies suggest that regulation of cortistatin-expressing interneurons by activity-dependent BDNF expression may contribute to regulation of sleep behavior.

  7. Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes

    OpenAIRE

    Groscurth Peter; Dumrese Claudia; Sundstrom Hanna; Walch Michael; Latinovic-Golic Sonja; Ziegler Urs

    2007-01-01

    Abstract Background Granulysin, a cytotoxic protein expressed in human natural killer cells and activated T lymphocytes, exhibits cytolytic activity against a variety of intracellular microbes. Expression and transcription have been partially characterised in vitro and four transcripts (NKG5, 519, 520, and 522) were identified. However, only a single protein product of 15 kDa was found, which is subsequently processed to an active 9 kDa protein. Results In this study we investigated generatio...

  8. Regulation of Plasminogen Activator Inhibitor-1 Expression by Tumor Suppressor Protein p53*

    OpenAIRE

    Shetty, Sreerama; Shetty, Praveenkumar; Idell, Steven; Velusamy, Thirunavukkarasu; Bhandary, Yashodhar P.; Shetty, Rashmi S.

    2008-01-01

    H1299 lung carcinoma cells lacking p53 (p53-/-) express minimal amounts of plasminogen activator inhibitor-1 (PAI-1) protein as well as mRNA. p53-/- cells express highly unstable PAI-1 mRNA. Transfection of p53 in p53-/- cells enhanced PAI-1 expression and stabilized PAI-1 mRNA. On the contrary, inhibition of p53 expression by RNA silencing in non-malignant human lung epithelial (Beas2B) cells decreased basal as well as urokinase-type plasminogen activator-induced PAI-...

  9. Differential activation of catalase expression and activity by PPAR agonists: Implications for astrocyte protection in anti-glioma therapy

    Directory of Open Access Journals (Sweden)

    Nicholas K.H. Khoo

    2013-01-01

    Full Text Available Glioma survival is dismal, in part, due to an imbalance in antioxidant expression and activity. Peroxisome proliferator-activated receptor (PPAR agonists have antineoplastic properties which present new redox-dependent targets for glioma anticancer therapies. Herein, we demonstrate that treatment of primary cultures of normal rat astrocytes with PPAR agonists increased the expression of catalase mRNA protein, and enzymatic activity. In contrast, these same agonists had no effect on catalase expression and activity in malignant rat glioma cells. The increase in steady-state catalase mRNA observed in normal rat astrocytes was due, in part, to de novo mRNA synthesis as opposed to increased catalase mRNA stability. Moreover, pioglitazone-mediated induction of catalase activity in normal rat astrocytes was completely blocked by transfection with a PPARγ-dominant negative plasmid. These data suggest that defects in PPAR-mediated signaling and gene expression may represent a block to normal catalase expression and induction in malignant glioma. The ability of PPAR agonists to differentially increase catalase expression and activity in normal astrocytes but not glioma cells suggests that these compounds might represent novel adjuvant therapeutic agents for the treatment of gliomas.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. Reductive Dehalogenase Gene Expression as a Biomarker for Physiological Activity of Dehalococcoides spp.

    OpenAIRE

    Lee, Patrick K. H.; David R. Johnson; Holmes, Victor F.; He, Jianzhong; Alvarez-Cohen, Lisa

    2006-01-01

    This study characterizes the transcriptional expression of the reductive dehalogenase (RDase)-encoding tceA and vcrA genes and evaluates their applicability as potential biological markers of Dehalococcoides activity. When Dehalococcoides ethenogenes 195 was provided with trichloroethene (TCE) as the electron acceptor, the expression of the tceA gene increased by 90-fold relative to that in cells starved of chlorinated ethenes, demonstrating that tceA gene expression is indicative of the acti...

  19. 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...

  20. 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...

  1. Activation of Six1 Expression in Vertebrate Sensory Neurons.

    Directory of Open Access Journals (Sweden)

    Shigeru Sato

    Full Text Available SIX1 homeodomain protein is one of the essential key regulators of sensory organ development. Six1-deficient mice lack the olfactory epithelium, vomeronasal organs, cochlea, vestibule and vestibuloacoustic ganglion, and also show poor neural differentiation in the distal part of the cranial ganglia. Simultaneous loss of both Six1 and Six4 leads to additional abnormalities such as small trigeminal ganglion and abnormal dorsal root ganglia (DRG. The aim of this study was to understand the molecular mechanism that controls Six1 expression in sensory organs, particularly in the trigeminal ganglion and DRG. To this end, we focused on the sensory ganglia-specific Six1 enhancer (Six1-8 conserved between chick and mouse. In vivo reporter assays using both animals identified an important core region comprising binding consensus sequences for several transcription factors including nuclear hormone receptors, TCF/LEF, SMAD, POU homeodomain and basic-helix-loop-helix proteins. The results provided information on upstream factors and signals potentially relevant to Six1 regulation in sensory neurons. We also report the establishment of a new transgenic mouse line (mSix1-8-NLSCre that expresses Cre recombinase under the control of mouse Six1-8. Cre-mediated recombination was detected specifically in ISL1/2-positive sensory neurons of Six1-positive cranial sensory ganglia and DRG. The unique features of the mSix1-8-NLSCre line are the absence of Cre-mediated recombination in SOX10-positive glial cells and central nervous system and ability to induce recombination in a subset of neurons derived from the olfactory placode/epithelium. This mouse model can be potentially used to advance research on sensory development.

  2. Activation of Six1 Expression in Vertebrate Sensory Neurons.

    Science.gov (United States)

    Sato, Shigeru; Yajima, Hiroshi; Furuta, Yasuhide; Ikeda, Keiko; Kawakami, Kiyoshi

    2015-01-01

    SIX1 homeodomain protein is one of the essential key regulators of sensory organ development. Six1-deficient mice lack the olfactory epithelium, vomeronasal organs, cochlea, vestibule and vestibuloacoustic ganglion, and also show poor neural differentiation in the distal part of the cranial ganglia. Simultaneous loss of both Six1 and Six4 leads to additional abnormalities such as small trigeminal ganglion and abnormal dorsal root ganglia (DRG). The aim of this study was to understand the molecular mechanism that controls Six1 expression in sensory organs, particularly in the trigeminal ganglion and DRG. To this end, we focused on the sensory ganglia-specific Six1 enhancer (Six1-8) conserved between chick and mouse. In vivo reporter assays using both animals identified an important core region comprising binding consensus sequences for several transcription factors including nuclear hormone receptors, TCF/LEF, SMAD, POU homeodomain and basic-helix-loop-helix proteins. The results provided information on upstream factors and signals potentially relevant to Six1 regulation in sensory neurons. We also report the establishment of a new transgenic mouse line (mSix1-8-NLSCre) that expresses Cre recombinase under the control of mouse Six1-8. Cre-mediated recombination was detected specifically in ISL1/2-positive sensory neurons of Six1-positive cranial sensory ganglia and DRG. The unique features of the mSix1-8-NLSCre line are the absence of Cre-mediated recombination in SOX10-positive glial cells and central nervous system and ability to induce recombination in a subset of neurons derived from the olfactory placode/epithelium. This mouse model can be potentially used to advance research on sensory development. PMID:26313368

  3. 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

  4. 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.

  5. 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.

  6. Telomerase activity, estrogen receptors (α, β), Bcl-2 expression in human breast cancer and treatment response

    International Nuclear Information System (INIS)

    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. 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. 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). 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

  7. 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...

  8. 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.

  9. 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.

  10. 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.

  11. 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...

  12. 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-positive c......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......-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...... surface-negative despite effective induction of apoptosis. Interestingly, inhibition of endolysosomes or normal ER/Golgi transport did not affect Hsp70 surface expression. Intracellular calcium and the transcription factor Sp1, which has been shown previously to be important for the intracellular stress...

  13. Ectopic Expression of an Activated RAC in Arabidopsis Disrupts Membrane CyclingD⃞V⃞

    OpenAIRE

    Bloch, Daria; Lavy, Meirav; Efrat, Yael; Efroni, Idan; Bracha-Drori, Keren; Abu-Abied, Mohamad; Sadot, Einat; Yalovsky, Shaul

    2005-01-01

    Rho GTPases regulate the actin cytoskeleton, exocytosis, endocytosis, and other signaling cascades. Rhos are subdivided into four subfamilies designated Rho, Racs, Cdc42, and a plant-specific group designated RACs/Rops. This research demonstrates that ectopic expression of a constitutive active Arabidopsis RAC, AtRAC10, disrupts actin cytoskeleton organization and membrane cycling. We created transgenic plants expressing either wild-type or constitutive active AtRAC10 fused to the green fluor...

  14. 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. PMID:23960123

  15. 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.

  16. Surviving gene expression in salivary adenoid cystic carcinoma%Survivin基因在涎腺腺样囊性癌中的表达意义

    Institute of Scientific and Technical Information of China (English)

    闫大勇

    2015-01-01

    目的 研究Survivin基因在涎腺腺样囊性癌中的表达,探索其在临床诊治中的意义.方法 采用免疫组化技术对病理科确诊的46例涎腺腺样囊性癌标本及20例正常涎腺标本进行Survivin基因表达的检测和分析.结果 正常涎腺组织标本均无Survivin基因表达,而73.91%的涎腺腺样囊性癌组织中Survivin基因呈阳性表达,差异有统计学意义(P<0.05).Survivin基因表达与患者年龄、性别、肿瘤分布部位、大小无相关性(P>0.05),而与肿瘤TNM分期、肿瘤病理类型、是否存在淋巴结转移呈正相关关系.结论 Survivin基因可作为涎腺腺样囊性癌早期诊断指标,其表达强度可作为判断预后的重要指标,针对Survivin基因开展靶向治疗与其他促凋亡治疗相结合的治疗方式将是临床诊治涎腺腺样囊性癌的重要途径.%Objective To investigate the expression of Survivin in salivary adenoid cystic carcinoma(SACC),in order to guide clinical diagnosis and treatment.Methods Immunohistochemical method was used to detect the expression of Survivin in 46 patients suffered with SACC and 20 normal parotid gland tissues.Results Survivin was not expressed in adjacent normal parotid gland tissues,but the positive rate of Survivin in SACC was 73.91%,there was significant difference(P <0.05).The positive expression of Survivin was not associated with ages,gender,site or tumor size,but showed positive correlation with TNM stages,histological classification and lymph node metastasis.Conclusions Survivin gene can be used as early diagnosis index of SACC,the overexpression of Survivin suggests the bad prognosis in SACC;Survivin gene may be identified as a defining diagnostic marker and a potential therapeutic target in SACC.

  17. Statins Activate Human PPAR Promoter and Increase PPAR mRNA Expression and Activation in HepG2 Cells

    Directory of Open Access Journals (Sweden)

    Makoto Seo

    2008-01-01

    Full Text Available Statins increase peroxisome proliferator-activated receptor (PPAR mRNA expression, but the mechanism of this increased PPAR production remains elusive. To examine the regulation of PPAR production, we examined the effect of 7 statins (atorvastatin, cerivastatin, fluvastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin on human PPAR promoter activity, mRNA expression, nuclear protein levels, and transcriptional activity. The main results are as follows. (1 Majority of statins enhanced PPAR promoter activity in a dose-dependent manner in HepG2 cells transfected with the human PPAR promoter. This enhancement may be mediated by statin-induced HNF-4. (2 PPAR mRNA expression was increased by statin treatment. (3 The PPAR levels in nuclear fractions were increased by statin treatment. (4 Simvastatin, pravastatin, and cerivastatin markedly enhanced transcriptional activity in 293T cells cotransfected with acyl-coenzyme A oxidase promoter and PPAR/RXR expression vectors. In summary, these data demonstrate that PPAR production and activation are upregulated through the PPAR promoter activity by statin treatment.

  18. The expression and function of microRNA-203 in lung cancer.

    Science.gov (United States)

    Jin, Jianhua; Deng, Jianzhong; Wang, Fang; Xia, Xiyi; Qiu, Tiefeng; Lu, Wenbin; Li, Xianwen; Zhang, Hua; Gu, Xiaoyan; Liu, Yungang; Cao, Weiguo; Shao, Wenlong

    2013-02-01

    We aimed to determine the expression of microRNA-203 (miR-203) in human lung cancer cell lines and to evaluate the effects of miR-203 by targeting survivin, on the lung cancer cell line 95-D to provide potential new strategies for treating lung cancer. The expression of miR-203 was detected using quantitative real-time PCR (qRT-PCR) in the in vitro cultured lung cancer cells A549, HCC827, NCI-H1299, and 95-D as well as in normal human bronchial epithelial cells. Following a 72-h transfection with the miR-203 precursor in 95-D lung cancer cells, the change in miR-203 expression was detected using qRT-PCR and the resulting effect on survivin protein expression was ascertained by Western blot analysis. The influence of miR-203 on the viability of 95-D lung cancer cells was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The effect of miR-203 on 95-D cell proliferation was analyzed using flow cytometry. The consequences of miR-203 expression on 95-D cell apoptosis were analyzed by Annexin V/propidium iodide double staining coupled with flow cytometry. The role of miR-203 in the invasive potential of 95-D cells was studied using a transwell chamber assay. A luciferase reporter gene system was used to verify that survivin is a target gene for miR-203. By qRT-PCR, the expression of miR-203 was lower in lung cancer cells than in normal bronchial epithelial cells (p transfection with the miR-203 precursor (p transfection with the miR-203 precursor, survivin protein levels in 95-D cells were significantly decreased (p transfection with the miR-203 precursor (p < 0.05). The expression of miR-203 is downregulated in lung cancer cells. miR-203 negatively regulates survivin protein expression and inhibits the proliferation and invasion of lung cancer cells. Therapeutic strategies that enhance miR-203 expression or silence survivin could potentially benefit lung cancer patients. PMID:23073851

  19. 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.

  20. 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.

  1. 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.

  2. Expression of Human Vascular Endothelial Growth Factor (VEGF165) in Pichia pastoris and Its Biological Activity

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective To express human vascular endothelial growth factor (hVEGF165) cDNA in Pichia pastroris, purify the expressed product and detect the biological activity of it. Methods  By inserting hVEGF165 cDNA coding 165 amino acid residues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides, a recombinant expression plasmid pPIC9K/hVEGF165 was constructed and transformed to yeast host strain KM71, then multiple-copy insert transformants were screened out and cultured in flasks, and hVEGF165 was expressed under the induction of 1% methanol. Results  SDS-PAGE showed that after being induced with 1% methanol for 4d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by Heparin-Sepharose CL6B affinity chromatography, the purity of expressed product reached above 90%. Biological assays proved that the expressed product could stimulate the proliferation of HUVEC. Conclusion  hVEGF165 was successfully expressed. The study opened up a wide prospect for the application of VEGF165 in the prevention and treatment of ischemic heart disease and other tissue ischemic diseases such as secondary arterial occlusion in limbs.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. relA over-expression reduces tumorigenicity and activates apoptosis in human cancer cells

    OpenAIRE

    Ricca, A; Biroccio, A; Trisciuoglio, D; M. Cippitelli; Zupi, G.; Bufalo, D Del

    2001-01-01

    We previously demonstrated that bcl-2 over-expression increases the malignant behaviour of the MCF7 ADR human breast cancer cell line and enhances nuclear factor-kappa B (NF-k B) transcriptional activity. Here, we investigated the direct effect of increased NF-k B activity on the tumorigenicity of MCF7 ADR cells by over-expressing the NF-k B subunit relA/p65. Surprisingly, our results demonstrated that over-expression of relA determines a considerable reduction of the tumorigenic ability in n...

  8. 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...

  9. 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

  10. Physical activity and prostate gene expression in men with low-risk prostate cancer

    OpenAIRE

    Magbanua, MJM; Richman, EL; Sosa, EV; Jones, LW; Simko, J; Shinohara, K.; Haqq, CM; Carroll, PR; Chan, JM

    2014-01-01

    Purpose: Vigorous physical activity after diagnosis of localized prostate cancer may reduce the risk of disease progression and prostate cancer-specific mortality. The molecular mechanisms by which physical activity may exert protective effects in the prostate remain unknown. Methods: We examined the associations between self-reported physical activity and gene expression patterns in morphologically normal prostate tissue of 71 men with low-risk prostate cancer on active surveillance. Differe...

  11. The protein kinase C inhibitor enzastaurin exhibits antitumor activity against uveal melanoma.

    Directory of Open Access Journals (Sweden)

    Xinqi Wu

    Full Text Available GNAQ mutations at codon 209 have been recently identified in approximately 50% of uveal melanomas (UM and are reported to be oncogenic through activating the MAPK/Erk1/2 pathway. Protein kinase C (PKC is a component of signaling from GNAQ to Erk1/2. Inhibition of PKC might regulate GNAQ mutation-induced Erk1/2 activation, resulting in growth inhibition of UM cells carrying GNAQ mutations. UM cells carrying wild type or mutant GNAQ were treated with the PKC inhibitor enzastaurin. Effects on proliferation, apoptosis, and signaling events were evaluated. Enzastaurin downregulated the expression of several PKC isoforms including PKCβII PKCθ, PKCε and/or their phosphorylation in GNAQ mutated cells. Downregulation of these PKC isoforms in GNAQ mutated cells by shRNA resulted in reduced viability. Enzastaurin exhibited greater antiproliferative effect on GNAQ mutant cells than wild type cells through induction of G1 arrest and apoptosis. Enzastaurin-induced G1 arrest was associated with inhibition of Erk1/2 phosphorylation, downregulation of cyclin D1, and accumulation of cyclin dependent kinase inhibitor p27(Kip1. Furthermore, enzastaurin reduced the expression of antiapoptotic Bcl-2 and survivin in GNAQ mutant cells. Inhibition of Erk1/2 phosphorylation with a MEK specific inhibitor enhanced the sensitivity of GNAQ wild type cells to enzastaurin, accompanied by p27(Kip1 accumulation and/or inhibition of enzastaurin-induced survivin and Bcl-2 upregulation. PKC inhibitors such as enzastaurin have activity against UM cells carrying GNAQ mutations through inhibition of the PKC/Erk1/2 pathway and induction of G1 arrest and apoptosis. Inhibition of the PKC pathway provides a basis for clinical investigation in patients with UM.

  12. 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.

  13. 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.

  14. Morphological expression of active tectonics in the Southern Alps

    Science.gov (United States)

    Robl, Jörg; Heberer, Bianca; Neubauer, Franz; Hergarten, Stefan

    2015-04-01

    Evolving drainage pattern and corresponding metrics of the channels (e.g. normalized steepness index) are sensitive indicators for tectonic or climatic events punctuating the evolution of mountain belts and their associated foreland basins. The analysis of drainage systems and their characteristic properties represents a well-established approach to constrain the impact of tectonic and climatic drivers on mountainous landscapes in the recent past. The Southern Alps (SA) are one of the seismically most active zones in the periphery of northern Adria. Recent deformation is caused by the ongoing convergence of the Adriatic and European plate and is recorded by numerous earthquakes in the domain of the SA. Deformation in the SA is characterized by back-thrusting causing crustal thickening and should therefore result in uplift and topography formation. The vertical velocity field determined by GPS-data clearly indicates a belt of significant uplift in the south South alpine indenter between Lake Garda in the west and the Triglav in the east and strong subsidence of the foreland basin surrounding the Mediterranean Sea near Venice, although subsidence is often related to ongoing subduction of the Adriatic microplate underneath Appennines. Despite of these short term time series, timing, rates and drivers of alpine landscape evolution are not well constrained and the linkage between crustal deformation and topographic evolution of this highly active alpine segment remains unclear for the following reasons: (1) The eastern Southern Alps were heavily overprinted by the Pleistocene glaciations and tectonic signals in the alpine landscape are blurred. Only the transition zone to the southern foreland basin remained unaffected and allows an analysis of a glacially undisturbed topography. (2) The major part of this domain is covered by lithology (carbonatic rocks) which is unsuitable for low temperature geochronology and cosmogenic isotope dating so that exhumation and erosion

  15. Co-targeting EGFR and survivin with a bivalent aptamer-dual siRNA chimera effectively suppresses prostate cancer.

    Science.gov (United States)

    Liu, Hong Yan; Yu, Xiaolin; Liu, Haitao; Wu, Daqing; She, Jin-Xiong

    2016-01-01

    Current targeted therapies using small kinase inhibitors and antibodies have limited efficacy in treating prostate cancer (PCa), a leading cause of cancer death in American men. We have developed a novel strategy by engineering an RNA-based aptamer-siRNA chimera, in which a bivalent aptamer specifically binds prostate-specific membrane antigen (PSMA) via an antibody-like structure to promote siRNA internalization in PCa cells, and two siRNAs specific to EGFR and survivin are fused between two aptamers. The chimera is able to inhibit EGFR and survivin simultaneously and induce apoptosis effectively in vitro and in vivo. In the C4-2 PCa xenograft model, the treatment with the chimera significantly suppresses tumor growth and angiogenesis. The inhibition of angiogenesis is mediated by an EGFR-HIF1α-VEGF-dependent mechanism. Our results support that the bivalent aptamer-driven delivery of two siRNAs could be a new combination therapeutic strategy to effectively inhibit multiple and conventionally "undruggable" targets. PMID:27456457

  16. Apoptosis of rat lens epithelial cells after eye contusion and effects of Survivin gene on apoptosis%大鼠眼钝挫伤后晶状体上皮细胞的凋亡及Survivin基因对凋亡的影响

    Institute of Scientific and Technical Information of China (English)

    刘建伟; 赵桂秋; 杜兆东; 张丽娜; 张振华

    2012-01-01

    Objective To investigate the apoptotic characteristics of rat lens epithelial cells(LEC) after eye contusion,and study the effects of Survivin gene on the apoptosis of LEC. Methods Forty-five adult Wistar rats were chosen,5 cases were set as normal control group,and other 40 cases were randomized into group A,B,C,D,10 cases in each group. Contusion eyes were induced by dropping a steel ball of 20 g from a height of 20 cm in group A,B,C,D. The lens was extracted at 1 hour( group A) ,12 hours( group B) ,24 hours( group C) and 48 hours( group D) after eye trauma, the apoptosis of LEC and expression of Survivin gene were detected. Results No pathological apoptosis or expression of Survivin gene were found in group A or normal rats. Apoptosis and expression of Survivin were observed in group B, C and D, the apoptotic ratewere(14.54?.98) % ,(41. 54 ?.07) % and(25.38 ?. 72) % .respectively,there were statistical differences between group C and group B, D (t = 9. 19,6. 64, both P ? 0. 05 ). The relative expressions of Survivin gene in group B, C, D were 0. 357 ?. 105, 0. 582 ?.131 and 0. 386 ?0.143,respectively. The relative expression of Survivin gene and apoptotic rate has a negative correlation in group B,C and D( r = -0.795, - 0.806, - 0.832, both P ? 0. 05). Conclusion The apoptosis of rat LEC and expression of Survivin gene can be observed after eye contusion. Apoptotic peak appears at 24 hours after trauma. Survivin gene may have an anti-apoptosis effect on the apoptosis of LEC after eye contusion.%目的 检测大鼠眼钝挫伤后晶状体上皮细胞凋亡的特点,探讨Survivin基因对晶状体上皮细胞凋亡的影响.方法 45只成年Wistar大鼠随机选取5只作正常对照组,其余40只随机分为A组、B组、C组、D组4组,每组各10只.除正常对照组外,其余鼠均用20 g钢球打击一眼制造眼钝挫伤模型后,于1h(A组)、12 h(B组)、24h(C组)、48 h(D组)后摘出晶状体,检测晶状体上皮细胞的凋亡及Survivin

  17. Activation of OX40 Augments Th17 Cytokine Expression and Antigen-Specific Uveitis

    NARCIS (Netherlands)

    Zhong, Wenwei; Hinrichs, David; Wu, Xiumei; Weinberg, Andrew; Hall, Mark; Spencer, Doran; Wegmann, Keith; Rosenbaum, James T.

    2010-01-01

    Uveitis is a major and common cause of visual disability Recent studies have shown that Th17 cells are implicated in the pathogenesis of this serious intraocular disorder Activated T cells express an inducible costimulatory molecule called OX40 and OX40 in turn promotes the activation and proliferat

  18. 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

  19. Activity-dependent modulation of odorant receptor gene expression in the mouse olfactory epithelium.

    Directory of Open Access Journals (Sweden)

    Shaohua Zhao

    Full Text Available Activity plays critical roles in development and maintenance of the olfactory system, which undergoes considerable neurogenesis throughout life. In the mouse olfactory epithelium, each olfactory sensory neuron (OSN stably expresses a single odorant receptor (OR type out of a repertoire of ∼1200 and the OSNs with the same OR identity are distributed within one of the few broadly-defined zones. However, it remains elusive whether and how activity modulates such OR expression patterns. Here we addressed this question by investigating OR gene expression via in situ hybridization when sensory experience or neuronal excitability is manipulated. We first examined the expression patterns of fifteen OR genes in mice which underwent neonatal, unilateral naris closure. After four-week occlusion, the cell density in the closed (sensory-deprived side was significantly lower (for four ORs, similar (for three ORs, or significantly higher (for eight ORs as compared to that in the open (over-stimulated side, suggesting that sensory inputs have differential effects on OSNs expressing different OR genes. We next examined the expression patterns of seven OR genes in transgenic mice in which mature OSNs had reduced neuronal excitability. Neuronal silencing led to a significant reduction in the cell density for most OR genes tested and thinner olfactory epithelium with an increased density of apoptotic cells. These results suggest that sensory experience plays important roles in shaping OR gene expression patterns and the neuronal activity is critical for survival of OSNs.

  20. 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.

  1. 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.

  2. 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

    Matrix metalloproteinases (MMPs) have been implicated in tissue damage associated with inflammatory bowel disease (IBD).As the role of the intestinal epithelium in this process is unknown, we determined MMP expression and enzyme activity in human colonic epithelial cells (CEC). MMP mRNA expression...... was assessed by reverse transcription-polymerase chain reaction in HT-29 and DLD-1 cells and in CEC isolated from biopsies from IBD and control patients. Total MMP activity in the cells was measured by a functional assay, based on degradation of a fluorescent synthetic peptide containing the specific bond...... 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...

  3. 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.

  4. Preparation of 99Tcm labeled survivin mRNA antisense PNA and gene imaging in nude mice bearing lung carcinoma A549 xenografts

    International Nuclear Information System (INIS)

    Objective: To prepare the 99Tcm-survivin mRNA antisense peptide nucleic acid (PNA)and investigate its value as a gene imaging agent in tumor bearing mice and early diagnosis in tumor. Methods: Survivin mRNA antisense PNA and mismatch PNA were synthesized. Four amino acids (Gly- (D)Ala-Gly-Gly) and Aba (4-aminobutyric acid) were linked to the 5' end of PNA. Gly- (D)Ala-Gly-Gly served as a chelating moiety for strong chelation of 99Tcm and Aba acted as a spacer to minimize the steric hindrance. PNAs were labeled with 99Tcm by the ligand-exchange method. The labeling efficiency and radiochemical purity were measured by HPLC and ITLC methods. There were five BALB/c nude mice bearing human lung carcinoma (A549) in each of antisense PNA and mismatch PNA groups. Gene imaging of 99Tcm-survivin mRNA antisense and mismatch PNAs were performed at 1, 2 and 4 h post the injection, respectively, and the T/NT ratio was measured by the method of ROI. The statistical comparisons of average values were performed with the two-group t-test for independent sample by SPSS 13.0. Results: The product kept stable in vitro. The labeling efficiency of 99Tcm-survivin mRNA antisense PNA was (95.48 ±1.92)% and more than 85% after the incubation for 24 h in serum. The radiochemical purity was >95%. The labeling efficiency of mismatch PNA was similar to the antisense PNA. 99Tcm-survivin mRNA antisense PNA was especially uptaken by tumor lesion, and its accumulation reached the top at 4 h post the injection. T/NT ratios at 1, 2, and 4 h were 2.70 ± 0.28, 3.44 ± 0.35,4.21 ± 0.63, respectively. In the comparison, the T/NT ratio of 99Tcm-survivin mRNA mismatch PNA at 4 h (3.12 ±0.50) was significantly lower (t=2.918, P=0.019). Conclusions: 99Tcm-survivin mRNA antisense PNA has high labeling efficiency,good stability and no need of purification. Its characteristic of especial uptake by tumor lesion provides the potential value in early diagnosis of tumor. (authors)

  5. 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.

  6. Cyclic strain increases protease-activated receptor-1 expression in vascular smooth muscle cells

    Science.gov (United States)

    Nguyen, K. T.; Frye, S. R.; Eskin, S. G.; Patterson, C.; Runge, M. S.; McIntire, L. V.

    2001-01-01

    Cyclic strain regulates many vascular smooth muscle cell (VSMC) functions through changing gene expression. This study investigated the effects of cyclic strain on protease-activated receptor-1 (PAR-1) expression in VSMCs and the possible signaling pathways involved, on the basis of the hypothesis that cyclic strain would enhance PAR-1 expression, reflecting increased thrombin activity. Uniaxial cyclic strain (1 Hz, 20%) of cells cultured on elastic membranes induced a 2-fold increase in both PAR-1 mRNA and protein levels. Functional activity of PAR-1, as assessed by cell proliferation in response to thrombin, was also increased by cyclic strain. In addition, treatment of cells with antioxidants or an NADPH oxidase inhibitor blocked strain-induced PAR-1 expression. Preincubation of cells with protein kinase inhibitors (staurosporine or Ro 31-8220) enhanced strain-increased PAR-1 expression, whereas inhibitors of NO synthase, tyrosine kinase, and mitogen-activated protein kinases had no effect. Cyclic strain in the presence of basic fibroblast growth factor induced PAR-1 mRNA levels beyond the effect of cyclic strain alone, whereas no additive effect was observed between cyclic strain and platelet-derived growth factor-AB. Our findings that cyclic strain upregulates PAR-1 mRNA expression but that shear stress downregulates this gene in VSMCs provide an opportunity to elucidate signaling differences by which VSMCs respond to different mechanical forces.

  7. 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.

  8. 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

  9. 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

  10. 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

  11. Expression

    Directory of Open Access Journals (Sweden)

    Wang-Xia Wang

    2014-02-01

    Full Text Available The miR-15/107 family comprises a group of 10 paralogous microRNAs (miRNAs, sharing a 5′ AGCAGC sequence. These miRNAs have overlapping targets. In order to characterize the expression of miR-15/107 family miRNAs, we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members, and other selected miRNAs, in 11 human tissues obtained at autopsy including the cerebral cortex, frontal cortex, primary visual cortex, thalamus, heart, lung, liver, kidney, spleen, stomach and skeletal muscle. miR-103, miR-195 and miR-497 were expressed at similar levels across various tissues, whereas miR-107 is enriched in brain samples. We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations (enriched for cortical neurons, astrocytes and microglia, respectively. In primary cultures of rat brain cells, several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system (CNS. In addition to mature miRNAs, we also examined the expression of precursors (pri-miRNAs. Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors. In summary, we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.

  12. 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.

  13. Cytokine, Chemokine and Immune Activation Pathway Profiles in Celiac Disease: An Immune System Activity Screening by Expression Macroarrays

    Directory of Open Access Journals (Sweden)

    José A. Garrote

    2008-01-01

    Full Text Available The aims of the study were to assess the usefulness of expression macroarrays to determine the pattern of expression of cytokines, chemokines and molecules related to immune system activation pathways, in non-stimulated intact intestinal tissue specimens from patients with active CD (aCD and on a gluten-free diet (GFD, to compare it with two groups of controls with either normal or altered mucosal architecture, and to establish putative targets for diagnostic markers or therapeutic intervention. We have experienced the lack of sensitivity to detect signal of genes with low level of expression. In spite of that, active CD seems to show a Th1 cytokine pattern, but with signs of Th2 activity. Cytokines such as IL-9, IL-11, IL-21 or MIF might be involved in mucosal inflammation in CD. In GFD, some memory cells and DC’s activity remains, and factors that maintain this remnant activation might be responsible of the fast mucosal response on gluten challenge. STAT3 and STAT5 pathways, and their regulatory molecules SOCS’s may result keys for understanding mucosal inflammation in gut and putative targets for further research.

  14. 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

  15. Soluble Expression and Characterization of Biologically Active Bacillus anthracis Protective Antigen in Escherichia coli

    OpenAIRE

    Nagendra Suryanarayana; Vanlalhmuaka,; Bharti Mankere; Monika Verma; Kulanthaivel Thavachelvam; Urmil Tuteja

    2016-01-01

    Bacillus anthracis secretory protein protective antigen (PA) is primary candidate for subunit vaccine against anthrax. Attempts to obtain large quantity of PA from Escherichia coli expression system often result in the formation of insoluble inclusion bodies. Therefore, it is always better to produce recombinant proteins in a soluble form. In the present study, we have obtained biologically active recombinant PA in small scale E. coli shake culture system using three different expression cons...

  16. Regulation of murine macrophage Ia-antigen expression by products of activated spleen cells

    OpenAIRE

    1980-01-01

    This investigation examined the effects of mediators derived form activated spleen cells on macrophage Ia-antigen expression and function. Incubation of adherent thioglycollate-induced murine peritoneal macrophages(> 90% Ia-) with concanavalin A (Con A)- stimulated spleen cell supernate (Con A sup) resulted in a dose- dependent increase in the percentage of Ia-containing (Ia+) phagocytic cells, as detected by antiserum-and-complement-mediated cytotoxicity. The Ia-antigen expression of macroph...

  17. Transcriptional activity, chromosomal distribution and expression effects of transposable elements in Coffea genomes.

    Directory of Open Access Journals (Sweden)

    Fabrício R Lopes

    Full Text Available Plant genomes are massively invaded by transposable elements (TEs, many of which are located near host genes and can thus impact gene expression. In flowering plants, TE expression can be activated (de-repressed under certain stressful conditions, both biotic and abiotic, as well as by genome stress caused by hybridization. In this study, we examined the effects of these stress agents on TE expression in two diploid species of coffee, Coffea canephora and C. eugenioides, and their allotetraploid hybrid C. arabica. We also explored the relationship of TE repression mechanisms to host gene regulation via the effects of exonized TE sequences. Similar to what has been seen for other plants, overall TE expression levels are low in Coffea plant cultivars, consistent with the existence of effective TE repression mechanisms. TE expression patterns are highly dynamic across the species and conditions assayed here are unrelated to their classification at the level of TE class or family. In contrast to previous results, cell culture conditions per se do not lead to the de-repression of TE expression in C. arabica. Results obtained here indicate that differing plant drought stress levels relate strongly to TE repression mechanisms. TEs tend to be expressed at significantly higher levels in non-irrigated samples for the drought tolerant cultivars but in drought sensitive cultivars the opposite pattern was shown with irrigated samples showing significantly higher TE expression. Thus, TE genome repression mechanisms may be finely tuned to the ideal growth and/or regulatory conditions of the specific plant cultivars in which they are active. Analysis of TE expression levels in cell culture conditions underscored the importance of nonsense-mediated mRNA decay (NMD pathways in the repression of Coffea TEs. These same NMD mechanisms can also regulate plant host gene expression via the repression of genes that bear exonized TE sequences.

  18. Inhibition of Ly-6A antigen expression prevents T cell activation

    OpenAIRE

    1990-01-01

    Antisense oligonucleotides complementary to the 5' end of the mRNA encoding the Ly-6A protein were used to block the expression of that protein. Using this approach we could inhibit the expression of Ly-6A by 60-80% in antigen-primed lymph node (LN) T cells as well as in the D10 T cell clone. Inhibition of Ly-6 expression resulted in the inability to restimulate in vitro, antigen-primed T cells. It also blocked the activation of normal spleen cells by Con A, monoclonal antibody (mAb) to CD3, ...

  19. 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.

  20. 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.

  1. Soluble Expression and Characterization of Biologically Active Bacillus anthracis Protective Antigen in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Nagendra Suryanarayana

    2016-01-01

    Full Text Available Bacillus anthracis secretory protein protective antigen (PA is primary candidate for subunit vaccine against anthrax. Attempts to obtain large quantity of PA from Escherichia coli expression system often result in the formation of insoluble inclusion bodies. Therefore, it is always better to produce recombinant proteins in a soluble form. In the present study, we have obtained biologically active recombinant PA in small scale E. coli shake culture system using three different expression constructs. The PA gene was cloned in expression vectors bearing trc, T5, and T7 promoters and transformed into their respective E. coli hosts. The growth conditions were optimized to obtain maximum expression of PA in soluble form. The expression construct PA-pET32c in DE3-pLysS E. coli host resulted in a maximum production of soluble PA (15 mg L−1 compared to other combinations. Purified PA was subjected to trypsin digestion and binding assay with lethal factor to confirm the protein’s functionality. Biological activity was confirmed by cytotoxicity assay on J774.1 cells. Balb/c mice were immunized with PA and the immunogenicity was tested by ELISA and toxin neutralization assay. This study highlights the expression of soluble and biologically active recombinant PA in larger quantity using simpler E. coli production platform.

  2. Soluble Expression and Characterization of Biologically Active Bacillus anthracis Protective Antigen in Escherichia coli.

    Science.gov (United States)

    Suryanarayana, Nagendra; Vanlalhmuaka; Mankere, Bharti; Verma, Monika; Thavachelvam, Kulanthaivel; Tuteja, Urmil

    2016-01-01

    Bacillus anthracis secretory protein protective antigen (PA) is primary candidate for subunit vaccine against anthrax. Attempts to obtain large quantity of PA from Escherichia coli expression system often result in the formation of insoluble inclusion bodies. Therefore, it is always better to produce recombinant proteins in a soluble form. In the present study, we have obtained biologically active recombinant PA in small scale E. coli shake culture system using three different expression constructs. The PA gene was cloned in expression vectors bearing trc, T5, and T7 promoters and transformed into their respective E. coli hosts. The growth conditions were optimized to obtain maximum expression of PA in soluble form. The expression construct PA-pET32c in DE3-pLysS E. coli host resulted in a maximum production of soluble PA (15 mg L(-1)) compared to other combinations. Purified PA was subjected to trypsin digestion and binding assay with lethal factor to confirm the protein's functionality. Biological activity was confirmed by cytotoxicity assay on J774.1 cells. Balb/c mice were immunized with PA and the immunogenicity was tested by ELISA and toxin neutralization assay. This study highlights the expression of soluble and biologically active recombinant PA in larger quantity using simpler E. coli production platform. PMID:26966576

  3. Inhibition of Nischarin Expression Promotes Neurite Outgrowth through Regulation of PAK Activity.

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    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.

  4. 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 .

  5. Adolescent Mouse Takes on An Active Transcriptomic Expression During Postnatal Cerebral Development

    Directory of Open Access Journals (Sweden)

    Wei Xu

    2014-06-01

    Full Text Available 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.

  6. Telomerase Activity and Telomerase Reverse Transcriptase Expression Induced by Selenium in Rat Hepatocytes

    Institute of Scientific and Technical Information of China (English)

    RI-AN YU; HUA-JIE CHEN; LING-FEI HE; BING CHEN; XUE-MIN CHEN

    2009-01-01

    Objectives To investigate the effects of sodium selenite on telomerase activity, apoptosis and expression of TERT, c-myc and p53 in rat hepatocytes. Methods Selenium at doses of 2.5, 5.0, and 10μmol/kg was given to SD rats by garage. In rat hepatocytes, telomerase activity was measured by the telomeric repeat amplification protocol (TRAP), apoptosis was detected by flow cytometry, and expressions of telomerase reverse transcriptase (TERT), c-myc and p53 were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).c-Myc and P53 proteins were detected by immunochemistry. Results Selenium at doses of 2.5, 5.0, and 10μmol/kg significantly increased hepatocellular telomerase activity and induced apoptosis in a close-dependent manner. Although selenium at doses of 2.5, 5.0, and 10μmol/kg displayed no obvious enhancing effect on the TERT mRNA expression in rat hepatocytes (P>0.05), it significantly increased the c-myc mRNA and p53 mRNA expression at the dose of 10μmol/kg (p<0.05). Selenium at doses of 5.0 and 10μmol/kg obviously increased the content of P53 protein in rat hepatocytes, but only at the dose of 10μmol/kg, it significantly promoted the value of c-Myc protein in them. Conclusion Selenium can slightly increase telomerase activity and TERT expression, and significantly induce apoptosis and over-expression of c-myc and p53 at relatively high doses. The beneficial effects of selenium on senescence and aging may be mediated by telomerase activation and expression of TERT, c-myc, and p53 in rat hepatocytes.

  7. Surface expression of Mo3e antigen by activated human monocytes and U-937 cells

    Energy Technology Data Exchange (ETDEWEB)

    Todd R.F. III; Bury, M.J.; Liu, D.Y.

    1986-03-05

    The surface expression of a protease-sensitive antigen, Mo3e, by activated human monocytes and U-937 cells is a plasma membrane feature of the activated state. Mo3e, which is an 80 kD protein on Western blot analysis, may represent the surface receptor for migration inhibitory factor (MIF), as evidenced by inhibition of MIF responsiveness produced by anti-Mo3e monoclonal antibody. Mo3e is barely detectable (by surface immunofluorescence) on freshly isolated monocytes but becomes expressed in high antigen density during 18-24 hrs culture in medium containing E. coli lipopolysaccharide (> 1 ng/ml), 4..beta..-phorbol 12-myristate 13-acetate (PMA) (5-10 nM), or muramyl dipeptide (0.1-1 ..mu..M). In U-937 cells, Mo3e surface expression is detectable after 24 hrs exposure to PMA and other pharmacological activators of protein kinase C: 4..beta..-phorbol 12, 13 dibutyrate, 4..beta..-phorbol 12, 13 didecanoate, mezerein, or Sn-1,2-dioctanoylglycerol. The biologically-inactivate phorbol compounds, 4..cap alpha..-phorbol 12, 13 didecanoate and 4/sub ..beta../-phorbol do not stimulate Mo3e expression. The calcium ionophore, ionomycin, has a synergistic effect on Mo3e expression stimulated by PMA; conversely, calcium antagonists block PMA-induced Mo3e expression. These results suggest the involvement of protein kinase C activation and intracellular calcium mobilization in the stimulated expression of Mo3e by activated human mononuclear phagocytes.

  8. 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. T