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pcDNATM6.2-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。结果干扰载体有效地阻断了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。
Objective To explore the inhibitory effects of RNAi targeting survivin to suppress the expression of survivin and to induce the apoptosis of human renal cancer cell line 786-0. Methods The target gene fragments were cloned into peDNATM 6.2-GW/EmGFPmiR vectors, and then the recombinant eukaryotic expression vectors were transfected into 786-O ceils with liposomes. The expression changes in survivin mRNA and protein were detected by RT-PCR and immunohistochemical assay, and the proliferation inhibitory rate （IR） of 786-O cells was detected by MTT assay. Results The mRNA and protein expressions of survivin in the siRNA group were growth and effectively cells. This significantly inhibited by the eukaryotic expression vectors, and it also inhibited significantly the proliferation of 786-O cells. Conclusion Silencing survivin gene by the RNAi technique can not only decrease the expressions of survivin gene and protein, but also inhibit the cell proliferation in 786-O may provide the experimental evidence for the survivin gene as a target of gene therapy in renal cancer.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第24期2444-2447,共4页
Journal of Third Military Medical University
