Department of Urology, Second Affiliated Hospital, School of Medicine, Zhejiang University, Jiefang Rd#88, Hangzhou, Zhejiang Province 310009, China.
Pathol Res Pract. 2009;205(12):823-7. doi: 10.1016/j.prp.2009.06.018. Epub 2009 Jul 22.
We investigated the inhibitory effect of small interfering RNA (siRNA) targeting survivin gene on cell proliferation and apoptosis in human renal clear cell carcinoma 786-O cells. qRT-PCR, immunocytochemistry, and Western Blot were used to detect Survivin gene expression in 786-O cells. Cell proliferation was determined by BrdU assay and PCNA expression. Cell apoptosis was analyzed through detection of caspase-3 activity, and the effect of survivin-siRNA on Bcl-2 gene expression was also examined. Forty-eight hours after transfection, survivin expression was markedly inhibited at the mRNA and protein level. Downregulation of survivin resulted in a significant inhibition of tumor cell growth. Caspase-3 activity showed that siRNA targeting survivin gene induced cell apoptosis in 786-O cells. Moreover, Bcl-2 protein expression was markedly inhibited by transfection with siRNA against survivin. These results indicate that siRNA targeting survivin gene can downregulate survivin gene expression in 786-O cells, inhibit growth, and induce apoptosis of renal carcinoma cells.
我们研究了针对生存素基因的小干扰 RNA（siRNA）对人肾透明细胞癌细胞 786-O 细胞增殖和凋亡的抑制作用。qRT-PCR、免疫细胞化学和 Western Blot 用于检测 786-O 细胞中生存素基因的表达。通过 BrdU 测定和 PCNA 表达来测定细胞增殖。通过检测 caspase-3 活性分析细胞凋亡，并检测 survivin-siRNA 对 Bcl-2 基因表达的影响。转染 48 小时后，mRNA 和蛋白水平上的生存素表达明显受到抑制。下调生存素导致肿瘤细胞生长明显受到抑制。caspase-3 活性表明，针对生存素基因的 siRNA 诱导 786-O 细胞发生细胞凋亡。此外，转染针对生存素的 siRNA 明显抑制 Bcl-2 蛋白表达。这些结果表明，针对生存素基因的 siRNA 可以下调 786-O 细胞中的生存素基因表达，抑制生长并诱导肾癌细胞凋亡。
