RNAi沉默KPNA2对人膀胱癌细胞系5637转移潜能的影响

Effects of KPNA2 small interfering RNA on migration and invasion of human bladder cancer cell lines 5637

目的探讨RNA干扰技术(RNA interference,RNAi)沉默KPNA2(karyopherin a2)基因表达对人膀胱癌细胞系5637迁移和侵袭能力的影响。方法实验分为siRNA敲低组和阴性对照组。siRNA敲低组设计合成靶向KPNA2序列的小分子干扰RNA(small interfering RNA,siRNA),用LipofectaminTM2000方法瞬时转染膀胱癌细胞系5637;阴性对照组采用无关序列RNA采集。转染后48h,应用蛋白质印迹法检测KPNA2的蛋白表达;Transwell实验检测细胞的迁移和侵袭能力。结果蛋白质印迹法检测结果显示,膀胱癌5637细胞转染KPNA2特异性siRNA 48h,与阴性对照组相比较,敲低组细胞KPNA2蛋白表达水平下降92.1%,差异有统计学意义,P=0.004 3。Transwell实验检测结果显示,敲低组细胞迁移能力明显受到抑制,细胞抑制率为61.7%,差异有统计学意义,P=0.038;敲低组细胞侵袭能力也明显受到抑制,细胞抑制率为58.6%,差异有统计学意义,P=0.026。结论靶向KPNA2的特异siRNA能够下调KPNA2在膀胱癌5637细胞中的蛋白表达水平,有效抑制膀胱癌5637细胞的迁移和侵袭能力。以KPNA2为靶点的RNA干扰技术有望成为膀胱癌基因治疗的新方法。

OBJECTIVE To study the effects of karyopherin a2（KPNA2）small interfering RNA（siRNA）transfection on migration and invasion of human bladder cancer cell lines 5637.METHODS A pair of KPNA2 siRNA was designed and synthesized,in vitro transiently transfected into human bladder cancer cell lines 5637 using LipofectamineTM2000.Forty-eight hours after transfection,western blot was used to analyze the expression level of KPNA2 protein.Transwell experiment in vitro was used to detect the variation of migration and invasion for bladder cancer cell lines 5637 before and after transfection.RESULTS After specific KPNA2 siRNA transfection,comparing with negative control group,the expression level of KPNA2 protein in siRNA transfect group was significantly decreased by 92.1%（P=0.004 3）,and the ability of cells＇ migration was significantly suppressed and the inhibition rate of tumor cells was 61.7%（P=0.038）.Likewise,the ability of cells invasion was suppressed and the inhibition rate of tumor cells was 58.6%（P=0.026）.CONCLUSION The siRNA of targeting KPNA2 gene can decrease the expression of KPNA2 protein in human bladder cancer cell lines 5637,and effectively suppress the migration and invasion ability of 5637 cell lines.The suppression of KPNA2 expression by siRNA technology may be a promising strategy for the treatment of human bladder cancer.