RNA干扰靶向Survivin促进膀胱癌细胞株BIU-87细胞凋亡的研究

Enhancement of Apoptosis of BIU-87 Bladder Cancer Cell Line by RNAi Targeting Survivin

目的针对凋亡抑制因子Survivin应用RNA干扰技术(RNA interference,RNA)i抑制膀胱癌细胞株BIU-87细胞内源性Survivin基因的表达进而促进膀胱癌细胞株BIU-87细胞凋亡.方法构建重组质粒pshRNA-survivin1、pshRNA-surv-ivin2和pshRNA-survivin 3并转染BIU-87细胞株,应用免疫组化法检测细胞中Survivin蛋白的表达,通过噻唑蓝比色法(MTT)检测细胞生长增殖抑制率,利用流式细胞术检测细胞凋亡率.结果重组质粒转染BIU-87细胞后,Survivin蛋白表达明显下调,细胞凋亡率显著增加,细胞生长明显受抑.结论膀胱癌细胞高表达Survivin,RNA干扰特异性抑制Survivin表达,可以促进膀胱癌细胞凋亡和增殖活性明显下降.

Objective To apply the small interfering RNAs（siRNA） targeting Survivin to inhibitexpression of endogenous Survivin gene in BIU-87 bladder cancer cells and to increase the apoptosis of BIU-87cells.Methods The recombinant plasmid pshRNA-survivin1,pshRNA-survivin2 and pshRNA-survivin3were constructed and transfected into BIU-87 cells,respectively.The expression level of Survivin wasdetermined by immunohistochemistry.The proliferation of BIU-87 cells was detected by MTT method.Theapoptosis of BIU-87 cells was detected by flow cytometry analysis（FCM）.Results After transfection ofrecombinant Survivin shRNA plasmid,the expression of Survivin was downregulated significantly,the growth ofcells was inhibited,and the apoptosis rate of BIU-87 cells increased significantly.Conclusions Theexpression levels of Survivin are high in bladder cancer cells.RNAi can inhibit specifically the expression ofSurvivin,suppress the proliferation,and induce the apoptosis of bladdercancercells.