RNA干扰技术特异性抑制肾癌survivin表达的实验研究

Inhibition of survivin expression by RNA interference in renal cancer cell line 786-O

目的:体外转录合成survivin siRNA(Small interference RNA,siRNA),观察其在肾癌786-O细胞株中抑制survivin的表达情况。方法:体外转录合成3组survivin序列特异性双链RNA(Double-stranded RNA,dsRNA),转染786-O细胞株中,用RT-PCR和Western blot检测转染后survivin基因的mRNA和蛋白的表达,MTT法检测细胞活性。结果:有2组转染特异性siRNA的786-O细胞表达survivin mRNA及蛋白均下调,活性受到抑制,另一组效果不明显。结论:体外转录合成特异性survivin siRNA能有效抑制肾癌786-O细胞中survivin的表达,抑制细胞的活性,RNA干扰(RNA interfering,RNAi)技术可能为肾癌的基因治疗提供一种新策略。

Objective：To study the inhibition of survivin expression by small interference RNA（siRNA） synthesized in vitro in renal cancer cell line 786-O. Methods：Three groups of survivin siRNAs were chemically synthesized in vitro and then transfected into cell line 786-O. The expression of survivin mRNA and protein after transfection were detected by reverse transcription polymerase chain reaction （RT-PCR）and Western Blotting. The proliferation of cells was detected by MTT. Results：Two sequence specific siRNAs targeting survivin down-regulated the expression of survivn mRNA and protein. The proliferation of cells was inhibited after transfection. The third one was ineffective. Conclusion：Sequence specific siRNA targeting survivin can efficiently inhibit the survivin expression and cell proliferation in renal cancer cell line 786-O. The successful application of RNAi targeting survivin might extend the list of available gene therapeutic modalities in renal cancer.