应用RNAi技术沉默survivin基因对肾癌786-O细胞的影响.docVIP

作者简介：李戈，男，四川省泸州市人，汉族，硕士研究生，医师，主要从事肾肿瘤方面的研究。电话通讯作者：王明，电 收稿日期：2009-1-15；修回日期：2009-4-22. 编辑/栾嘉 应用RNAi技术沉默survivin基因对肾癌786-O细胞的影响 　　　　　　　　　　　　　　　 李戈，王明 （重庆医科大学附属第一医院泌尿外科，重庆　４０００１６） 【摘要】目的：运用RNA干扰(RNA interference，RNAi)技术特异性阻断肾癌786-O细胞系中survivin基因的表达，并研究survivin沉默后对786-O细胞的影响。 方法：用真核转录载体pcDNA?6.2-GW/EmGFPmiR构建针对survivin基因的干扰载体，用脂质体法转染肾癌786-O细胞，通过RT-PCR、免疫组化分别检测survivin基因的mRNA和蛋白水平的表达，MTT法检测细胞生长的影响。 结果：干扰载体有效的阻断了786-O细胞中survivin基因在mRNA与蛋白水平上的表达，细胞活性受到抑制。 结论： RNAi技术沉默survivin基因可以下调肾癌786-O细胞survivin基因的表达，抑制细胞的活性，为以survivin基因为靶点的肾癌基因治疗研究提供了实验依据。 【关键词】　RNA干扰 ； survivin ； 肾癌 ；基因治疗 【中图分类号】R737.1 【文献标识码】A Inhibition of human renal cancer cell line 786-O by silencing survivin gene with RNAi LI Ge , WANG Ming Department of Urinary Surgery, the First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China 【Abstract】 Objective：To observe the inhibitory effects of RNAi suppressing the expression of survivin and inducing the apoptosis of human renal cancer cell line 786-O．Methods：The target gene fragments were cloned into pcDNA?6.2-GW/EmGFPmiR vectors，and then the recombinant eukaryotic expression vectors were transfected into 786-O cells with liposomes． The expression changes in survivin mRNA and protein were detected by RT-PCR and immunohistochemisty，the proliferation inhibitory rate (IR) of 786-O cells was detected by MTT．Results： The mRNA and protein expressions of survivin gene in the siRNA group were significantly inhibited by the eukaryotic expression vectors，and it also inhibit significantly the growth and proliferation of 786-O cells．Conclusion：Silencing survivin gene by the RNAi technique can not only decrease effectively the expressions of survivin gene and protein，but also inhibit the cell proliferation in 786-O cells．This may provide the experimental evidence for the survivin gene as a target of gene therapy in renal canc