survivin 靶向siRNA 对膀胱癌细胞增殖和凋亡作用的体外研究

目的 观察靶向survivin的siRNA对膀胱癌细胞T-24增殖和凋亡的影响.方法 使用lipofectamineTM2000将靶向survivin的siRNA真核表达载体转染至膀胱癌细胞T-24,半定量RT-PCR检测T-24细胞survivin基因表达的变化;MTT法检测对T-24细胞增殖的影响;膜联蛋白V-PI(annexinV-PI)染色及流式细胞技术检测诱导凋亡的影响.结果 靶向survivin的序列特异性siRNA可以高效率抑制T-24细胞survivin基因在基因水平的表达,mRNA抑制率为61.73%;细胞重新接种24、48h后,其增殖抑制率分别为32.42%和37.48%;转染24h后可以诱导细胞凋亡16.76%.结论 利用RNA干扰技术沉默survivin基因表达可以显著抑制T-24细胞增殖,并在一定程度上诱导其自发凋亡,靶向survivin的RNA干扰技术在膀胱癌的基因治疗中具有一定价值.

Investigation of siRNA targeted to survivin in suppressing proliferation and inducing apoptosis in bladder cancer

Objective To investigate the effect of blocking the survivin expression with RNA interference(RNAi)techniques on supperresing proliferation and inducing apoptosis of bladder cancer T-24 cells.Methods A siRNA directed against survivin was transfeted into bladder cancer T-24 cells with lipofectamineTM2000.The changes of survivin expression were detected by semi-quantitive RT-PCR.The effect of suppressing proliferation of T-24 cell was detected by MTT assay.The effect of inducing T-24 cell apoptosis was detected by flow cytometry.Results The sequence-specific siRNA efficiently blocked the survivin expression at mRNA level.The expression inhibition rate was 61.73% at mRNA mevel detected by semi-quantitive RT-PCR.To blocking the survivin expression could suppress proliferration of T-24 cells significantly.At 24h and 48h after the cells were reseeded,the proliferation inhibition rates were 32.42% and 37.48%,and at 24h after transfection,apoptosis was induced in 16.76% of the cells as detected by flow cytometry assay.Conclusion Blocking the survivin expression with RNAi technology can significantly suppress proliferation of T-24 cells and induce apoptosis to a certain degree.RNAi targeted to survivin has a potential value in gene therapy of bladder cancer.