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

作　　者：严勇[1] 李克功[1] 詹辉[2] 丁明霞[2] 王春晖[2] 王剑松[2] 

机构地区：[1]昆明市第一人民医院泌尿外科,云南昆明650011 [2]昆明医学院第二附属医院泌尿外科云南省泌尿外科研究所,云南昆明650101

出　　处：《昆明医学院学报》2010年第11期8-12,共5页Journal of Kunming Medical College

基　　金：国家自然科学基金资助项目(30760251)

摘　　要：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.

关 键 词：RNA干扰 SURVIVIN 基因 膀胱肿瘤 

分 类 号：R246.5[医药卫生—针灸推拿学]