VEGF和Survivin在肾癌中的表达及其相关性研究

目的：探讨肾癌组织中血管内皮生长因子VEGF与凋亡抑制蛋白Survivin表达的相关性及其之间的关系,研究Survivin和VEGF在肾癌发生发展中的作用机制。方法：应用免疫组织化学方法检测70例肾癌组织和70例癌旁正常肾脏组织中VEGF和Survivin的表达,并将检测结果与临床病理特征进行综合分析。结果：VEGF和Survivin在肾癌中表达均高于癌旁正常肾脏组织;Survivin和VEGF在肾癌中的阳性表达率分别为75.71%（53/70）和72.86%（51/70）,在癌旁肾脏组织中的表达率分别为0%（0/70）、17.14%（12/70）,差异均有显著性意义（P〈0.05）;VEGF和Survivin的表达与患者的性别、年龄、肿瘤大小、病理分级均无相关性;VEGF和Survivin表达呈正相关性。结论：VEGF和Survivin在肾癌组织中表达率较高,为肾癌的分子靶向治疗提供了新的靶点。Survivin和VEGF在RCC中的表达关系密切,测定RCC中Survivin、VEGF蛋白的表达,有助于临床判断病人预后。     

VEGF和Survivin在肾癌中的表达及其相关性研究

目的:探讨肾癌组织中血管内皮生长因子VEGF与凋亡抑制蛋白Survivin表达的相关性及其之间的关系,研究Survivin和VEGF在肾癌发生发展中的作用机制。方法:应用免疫组织化学方法检测70例肾癌组织和70例癌旁正常肾脏组织中VEGF和Survivin的表达,并将检测结果与临床病理特征进行综合分析。结果:VEGF和Survivin在肾癌中表达均高于癌旁正常肾脏组织;Survivin和VEGF在肾癌中的阳性表达率分别为75.71%(53/70)和72.86%(51/70),在癌旁肾脏组织中的表达率分别为0%(0/70)、17.14%(12/70),差异均有显著性意义(P0.05);VEGF和Survivin的表达与患者的性别、年龄、肿瘤大小、病理分级均无相关性;VEGF和Survivin表达呈正相关性。结论:VEGF和Survivin在肾癌组织中表达率较高,为肾癌的分子靶向治疗提供了新的靶点。Survivin和VEGF在RCC中的表达关系密切,测定RCC中Survivin、VEGF蛋白的表达,有助于临床判断病人预后。     

大肠癌中Livin和Survivin的表达及相关性研究

目的探讨Livin和Survivin在大肠癌组织中的表达与临床病理因素间的关系以及两者间的相互关系。方法应用免疫组织化学S-P方法检测Livin和Survivin在65例大肠癌组织中的表达情况。结果 Livin和Survivin蛋白在大肠癌组织中的阳性表达率分别是72.3%(47/65)、73.8%(48/65),均明显高于大肠正常黏膜组织24%(6/25)、12%(3/25)(P0.05)。Livin和Survivin蛋白表达与肿瘤浸润深度、淋巴结转移及临床Duke's分期有关,与患者年龄、性别及肿瘤分化程度无关。Livin和Survivin蛋白在大肠癌组织的表达呈正相关。两者协同表达与肿瘤浸润深度、淋巴结转移情况以及临床Duke's分期呈正相关。结论 Livin和Survivin在大肠癌的发生发展中均具有重要作用,同时两者具有协同作用,提示联合检测较单一检测更有意义。     

Survivin 及Anx-A1 在肝癌组织中的表达及其临床意义

目的:探讨Survivin及Anx-A1在肝癌组织中的表达及其临床意义。方法:收集原发性肝癌病例45例,采用免疫组织化染色法检测Survivin及Anx-A1在肝癌组织及癌旁正常组织中的表达,分析Survivin及Anx-A1的表达与肝癌临床病理特征的关系。结果:Survivin在肝癌组织中的阳性表达率为86.67%,在癌旁正常组织中的阳性表达率为17.78%;Anx-A1在肝癌组织中的阳性表达率为46.67%,在癌旁正常组织中的阳性表达率为8.89%;Survivin及Anx-A1在肝癌组织中的阳性表达率均显著高于癌旁正常组织,差异具有统计学意义(P0.05);不同肿瘤分级患者肝癌组织中Survivin与Anx-A1的表达水平存在显著差异(P0.01),肿瘤分级越高,Survivin与Anx-A1表达水平越高。结论:Survivin及Anx-A1的表达与肝癌的发生发展密切相关,可用于肝癌的辅助诊断。     

胃癌组织中Survivin、P53蛋白表达和微血管密度测定及意义

目的探讨Survivin,P53蛋白在胃癌组织中的表达及其与血管生成的关系。方法用免疫组化方法对65例胃癌组织Survivin,P53,CD34的表达进行检测。结果胃癌组织中Survivin蛋白阳性表达率为70.77%（46/65）,P53蛋白阳性表达率为64.62%（42/65）。Survivin表达与分化程度和淋巴结转移有关（P〈0.05）,而与临床分期无关（P〉0.05）;P53的阳性表达与分化程度、临床分期和淋巴结转移有关（P〈0.05）。Survivin表达与P53的表达密切相关（r=0.413,P〈0.01）。Survivin阳性表达组织中的MVD值为103.04±15.06,阴性组为81.89±12.15,二者有显著差异（P〈0.01）。P53阳性表达胃癌组织中的MVD值为105.83±15.06,阴性组为81±12.71,二者有显著差异（P〈0.01）。结论Survivin和P53突变对凋亡的抑制的协同作用及其促进血管生成的作用在胃癌的发生发展中起关键作用。     

MMP-7和Survivin在结直肠癌中的表达及其临床意义

目的探讨Wnt信号通路中基质金属蛋白酶7（matrix melluoproteinases7,MMP-7）和凋亡抑制基因Survivin在结直肠癌组织中表达及其与临床病理特征的关系。方法应用免疫组织化学染色（Elivision）方法检测100例结直肠癌组织和60例癌旁正常黏膜组织中MMP-7和Survivin蛋白的表达。结果MMP-7蛋白在100例结直肠癌组织和60例癌旁正常黏膜组织中的阳性表达率分别为77．00％（77／100）和13．33％（8／60）,两组问差异有统计学意义（P〈0．01）;Survivin蛋白在100例结直肠癌组织和60例癌旁正常黏膜组织中的阳性表达率分别为65．00％（65／lOO）和15．00％（9／60）,两组问差异有统计学意义（P〈0．01）。MMP-7与Survivin蛋白阳性表达均与肿瘤的淋巴结转移和Dukes分期有关（P〈0．05）,此外,MMP-7蛋白在结直肠癌中的阳性表达也与肿瘤的浸润深度有关（P〈0．05）。而MMP-7与Survivin蛋白的阳性表达无相关性（r=0．097,P〉O．05）。结论MMP-7和Survivin在结直肠癌中的高表达可能与结直肠癌的发生、发展、浸润和转移等相关,检测癌组织中MMP-7和Survivin的表达有助于为结直肠癌的病情进展及预后判断提供帮助。     

应用组织芯片技术研究Survivin基因蛋白在前列腺肿瘤组织中的表达及其意义

目的:应用组织芯片技术分析Survivin基因蛋白在人类前列腺癌组织、前列腺正常组织及前列腺良性痛变组织中的表达情况.方法:采用兔抗人survivin单克隆抗体的免疫组织化学ABC法,研究Survivin在不同前列腺组织的表达,并分析Survivin在不同前列腺组织中的表达差异.结果:免疫组化结果显示,前列腺癌组织与前列腺良性病变组织及正常前列腺组织中Survivin的表达相比呈显著性差异(P<0.05).结论:Survivin在前列腺癌组织中呈高表达,提示其可能对前列腺癌的发生或发展有重要作用.     

Survivin在大肠癌中的表达及其与Bcl-2、P53的关系

目的:研究凋亡抑制因子Survivin蛋白在大肠癌中的表达及其与患者临床病理和预后的关系。检测大肠癌组织中Survivin的表达与Bcl-2,P53的表达的关系。方法:用免疫组织化学方法检测115例大肠癌组织中Survivin,Bcl-2及P53的蛋白表达,并对病例随访5年。结果:Survivin在大肠癌中的阳性表达率为74/115(64．3%)。正常肠黏膜组织不表达Survivin。Survivin与患者的临床病理特征无显著相关性(P＞0．05)。Survivin的表达率在Bcl-2阳性的患者明显高于Bcl-2阴性的患者(P＜0．001),但是和P53的表达无显著相关性(P＞0．05)。Survivin阳性的患者5年生存率明显低于Survivin阴性的患者(P=0．001)。结论:在大肠癌组织中检测Survivin对于肿瘤患者的预后以及基于抗凋亡机制的肿瘤靶向治疗都有很重要的意义。     

COX-2和Survivin在B细胞非霍奇金淋巴瘤中的表达及意义

目的:探讨COX-2和Survivin在B细胞非霍奇全淋巴瘤(B-NHL)中的表达及临床意义.方法:采用免疫组化SP法检测43例B-NHL和10例良性淋巴结病变组织中COX-2和Survivin的表达.结果:COX-2和Survivin在B-NHL中的阳性表达率分别为55.8%(24/43)和69.8%(30/43),与对照组相比差异具有显著性(P＜0.05.Ⅲ/Ⅳ期B-NHL患者COX-2蛋白的表述要高于Ⅰ/Ⅱ期患者(P＜0.05).Survivin的表达与B-NHL的组织病理学等级和国际预后指数(IPL)具有相关性(P＜0.05).Spearman相关分析表明COX-2的表达与Survivin的表达呈正相关(r=1.000,P=0.030).结论:COX-2和Survivin在B-NHL中表达上调以及两者之间的阳性表达密切相关,表明COX-2和Survivin在B-NHL的发生和发展中具有协同作用.     

Survivin的研究现状与RNA干扰在基因治疗中的进展

Survivin是新近发现的凋亡抑制蛋白,在正常组织中不表达而在恶性肿瘤中高表达,与肿瘤的预后、复发、转移关系密切,具有调控细胞周期,凋亡和增殖的作用;还与对化疗的耐药性和放疗的敏感性有关;此外,与肿瘤新生血管的形成关系也十分密切.针对Survivin这些特性,应用新的基因治疗技术-RNA干扰技术降低Survivin的表达可对肿瘤组织的凋亡和增殖产生影响,由于RNA干扰技术具有的高效性,特异性,长效性,使它成为肿瘤基因治疗的重要方法,应用RNA干扰技术针对Survivin靶位降低Survivin的表达有望成为今后肿瘤基因治疗的方向.     

Correlation between the levels of survivin and survivin promoter-driven gene expression in cancer and non-cancer cells

Survivin, a member of the inhibitor of apoptosis (IAP) protein family, is associated with malignant transformation and is over-expressed in most human tumors. Using lipoplex-mediated transfection, we evaluated the activity of the reporter enzyme, luciferase, expressed from plasmids encoding the enzyme under the control of either the cytomegalovirus (CMV) or survivin promoters, in tumor- and non-tumor-derived human and murine cells. We also examined whether there is a correlation between the survivin promoter-driven expression of luciferase and the level of endogenous survivin. Human cancer cells (HeLa, KB, HSC-3, H357, H376, H413), oral keratinocytes, GMSM-K, and chemically immortalized human mammary cells, 184A-1, were transfected with Metafectene at 2 μl/1 μg DNA. Murine squamous cell carcinoma cells, SCCVII, mouse embryonic fibroblasts, NIH-3T3, and murine immortalized mammary cells, NMuMG, were transfected with Metafectene PRO at 2 μl/1 μg DNA. The expression of luciferase was driven by the CMV promoter (pCMV.Luc), the human survivin promoter (pSRVN.Luc-1430), or the murine survivin promoters (pSRVN.Luc-1342 and pSRVN.Luc-194). Luciferase activity was measured, using the Luciferase Assay System and expressed as relative light units (RLU) per ml of cell lysate or per mg of protein. The level of survivin in the lysates of human cells was determined by ELISA and expressed as ng survivin/mg protein. In all cell lines, significantly higher luciferase activity was driven by the CMV promoter than by survivin promoters. The expression of luciferase driven by the CMV and survivin promoters in murine cells was much higher than that in human cells. The cells displayed very different susceptibilities to transfection; nevertheless, high CMV-driven luciferase activity appeared to correlate with high survivin-promoter driven luciferase expression. The survivin concentration in lysates of cancer cells ranged from 5.8 ± 2.3 to 24.3 ± 2.9 ng/mg protein (mean, 13.7 ng/mg). Surprisingly, elevated survivin protein was determined in lysates of non-tumor-derived cells. Survivin levels for GMSM-K and 184A-1 cells, were 16.7 ± 8.7 and 13.5 ± 6.2 ng/mg protein, respectively. The expression of endogenous survivin did not correlate with the level of survivin promoter-driven transgene activity in the same cells. The expression of survivin by non-tumorigenic, transformed cell lines may be necessary for their proliferative activity. The level of survivin promoter-driven gene expression achieved via liposomal vectors in OSCC cells was too low to be useful in cancer-cell specific gene therapy.     

Survivin expressions in human hepatoma HepG2 cells exposed to ionizing radiation of different LET

Survivin is a member of the inhibitors of apoptosis (IAP) protein family that interferes with post-mitochondrial events including activation of caspases. To examine the regulation of survivin expression in response to irradiation with different linear energy transfer (LET), human hepatoma HepG2 cells were irradiated in vitro with X-rays and carbon ions. Cellular sensitivities to low- and high-LET radiation were determined by colony formation. Survivin expression at mRNA and protein level were measured with RT-PCR and Western blot analyses, respectively. Radiation-induced cell cycle arrest and apoptosis were investigated with flow cytometry. We found that low-LET X-rays induced dose-dependent increases in survivin expression. After exposure to high-LET carbon ions, survivin expression gradually increased from 0 to 4 Gy, and then declined at 6 Gy. More pronounced survivin expression, stronger G(2)/M phase arrest was observed after exposure to carbon ions in comparison with X-rays at doses from 0 to 4 Gy. These observations indicate that there is a differential survivin expression in response to different LET radiations and the cycle arrest mechanism may be associated with it. In addition, our data on induction of apoptosis are compatible with the assumption that survivin expression induced by low-LET X-rays radiation may play a critical role in inhibiting apoptosis. However, after irradiation with ions, an anti-apoptotic function of survivin is not evident, possibly because of the serious damage produced by densely ionizing radiation.     

Crystal structure of human survivin reveals a bow tie-shaped dimer with two unusual alpha-helical extensions

Survivin is a mitotic spindle-associated protein involved in linking mitotic spindle function to activation of apoptosis in mammalian cells. The structure of the full-length human survivin has been determined by X-ray crystallography to 2.7 A. Strikingly, the structure forms a very unusual bow tie-shaped dimer. It does not dimerize through a C-terminal coiled-coil, contrary to sequence analysis prediction. The C-terminal helices contain hydrophobic clusters with the potential for protein-protein interactions. The unusual shape and dimensions of survivin suggest it serves an adaptor function through its alpha-helical extensions.     

Survivin inhibition is critical for Bcl-2 inhibitor-induced apoptosis in hepatocellular carcinoma cells

Our study aims to study the therapeutic effects of a novel Bcl-2 inhibitor, ABT-263, on hepatocellular carcinoma (HCC) and to provide primary preclinical data for future clinical trial with ABT-263. In this study we showed that Bcl-xL and survivin were up-regulated in HCC cell lines and human liver cancer tissues. Clinic used ABT-263 single treatment had no apoptotic effects on HCC cells whereas higher doses of ABT-263 did. Interestingly, the combination treatment of ABT-263 with survivin inhibitor YM-155 could result in significant apoptosis in HCC cells. Survivin inhibition through gene silencing significantly enhanced ABT-263 to induce apoptosis in HCC cells. We found that low dose of ABT-263 single treatment resulted in ERK activation and survivin up-regulation, which might be involved in the resistance of HCC cells to ABT-263 since blockade of ERK activation sensitized ABT-263-induced apoptosis. Importantly, ABT-263 and YM-155 combination treatment had no apoptotic effects on normal human hepatocytes. Taken together, these data suggest the combination treatment of Bcl-2 inhibitor and survivin inhibition may have a great potential for liver cancer therapy.