Survivin基因对大肠癌细胞HT29生物学行为的影响  

Influence of Survivin gene on biological behaviours of colorectal cancer HT29 cells

作　　者：盛文文[1,2] 陆娜娜[1] 尹玉[1] 曹立宇[1] 

机构地区：[1]安徽医科大学基础医学院病理学教研室,合肥230032 [2]皖西卫生职业学院药学系化学教研室,六安237005

出　　处：《临床与实验病理学杂志》2013年第4期378-381,共4页Chinese Journal of Clinical and Experimental Pathology

基　　金：安徽省自然科学基金(090413098)

摘　　要：目的探讨Survivin基因对人大肠癌细胞HT29增殖及凋亡的影响。方法以脂质体Lip-2000为载体,用针对Survivin特异靶点的siRNA转染人大肠癌细胞HT29后,应用免疫细胞化学SP法、Western blot技术检测Survivin蛋白表达变化;采用MTT法检测细胞增殖;采用流式细胞技术检测细胞凋亡。结果免疫细胞化学结果:正常对照组、脂质体对照组及阴性错配对照组中Survivin均呈细胞质强阳性表达,Survivin-siRNA组细胞质呈弱阳性表达;Western blot检测结果:Survivin-siRNA组细胞的蛋白条带亮度明显低于正常对照组、脂质体对照组和阴性错配对照组;MTT检测结果:与阴性错配对照组相比,Survivin-siRNA组细胞生长出现明显的抑制(P<0.05)。不同时段(24、48、72 h)肿瘤细胞增殖抑制率之间差异有显著性(P<0.05);流式细胞术检测结果显示Survivin-siRNA组细胞凋亡比例显著高于空白对照组及阴性错配对照组(P<0.01)。结论 Sur-vivin基因参与调控大肠癌细胞的增殖和凋亡,Survivin有望成为大肠癌基因治疗的新靶点。Purpose To explore the influence of Survivin gene on the proliferation and apoptosis of HT29 cells. Methods Survivin- targeted siRNA was transfeted into human eolorectal cancer HT29 cells with Lip-2000 medium. Expression of survivin protein was de- tected by immunocytoehemistry and Western blot. Cell proliferation was measured by MTT assay. Cell apoptosis was detected by flow cytometry （FCM）. Results Immunocytoehemical results indicated that Survivin had strongly positive expression in the normal control group, Lip-2000 control group and mismatch control group, and showed weak expression in Survivin-siRNA interference group. West- ern blot indicated that brightness of protein bands in Survivin-siRNA interference group was significantly lower than those in normal control group, Lip-2000 control group and mismatch control group. Compared with control group, the inhibitory rate of cell proliferation was significantly increased in Survivin-siRNA interference group compared with negative mismatch group （ P 〈 0. 05 ）. Cell proliferation rates were significantly different between 24, 48 and 72 h after transfeetion （ P 〈 0. 05 ）. FCM showed that diploid peak appeared in Survivin-siRNA interference group and cell apoptosis rates of Survivin-siRNA interference group were obviously higher than those of empty control groups （ P 〈 0. 01 ）. Conclusion Survivin gene regulates the proliferation of colorectal cancer cells and induces apopto- sis, which may become a new target in gene therapy for colorectal cancer.

关 键 词：大肠肿瘤 SURVIVIN RNA干扰 细胞增殖 细胞凋亡 

分 类 号：R730.5[医药卫生—肿瘤] R735.3[医药卫生—临床医学]