腺病毒载体介导shRNA抑制生存素对PC-3细胞的影响  

Effect of survivin gene inhibition mediated by RNA interference in prostate cancer PC-3 cell line

作　　者：徐明[1] 单玉喜[1] 阳东荣[1] 薛波新[1] 

机构地区：[1]苏州大学附属第二医院泌尿外科,215004

出　　处：《江苏医药》2010年第13期1574-1577,共4页Jiangsu Medical Journal

基　　金：江苏省自然科学基金(BK2008167);2008年苏州市科技发展计划(社会发展及医药)项目

摘　　要：目的运用RNA干扰(RNAi)技术抑制雄激素非依赖前列腺癌细胞株(PC-3)中生存素的表达,检测生存素抑制对PC-3细胞的影响。方法构建负载生存素短发夹RNA(shRNA)且携带绿色荧光蛋白(GFP)的重组腺病毒,体外转染PC-3细胞。MTT法监测细胞增殖活性,流式细胞术检测凋亡率,RT-PCR和Western blot分别对生存素mRNA及其蛋白表达进行测定。结果负载生存素shRNA重组腺病毒构建成功;转染96h后,PC-3细胞增殖受到抑制,并可诱导出PC-3细胞凋亡,凋亡率达到(14.43±0.57)%,生存素mRNA及其蛋白的表达强度分别降低了(72.23±1.11)%和(71.75±2.64)%,与阴性对照组及空白对照组比较差异有统计学意义(P<0.01)。结论腺病毒载体介导的生存素shRNA可有效地抑制PC-3细胞中生存素的表达,RNAi结合腺病毒载体系统抑制生存素表达有望成为治疗雄激素非依赖前列腺癌的有效方式之一。Objective To evaluated the effect of survivin gene inhibition mediated by RNA interference in prostate cancer PC-3 cell line.Methods A recombinant adenovirus（pGsadeno-sur shRNA） bearing survivin short hairpin RNA（shRNA） and GFP was constructed and transfected into PC-3 cell line.MTT assay and flow cytometry were used to monitor the proliferation activity and apoptosis rate of PC-3 cell after transfection.RT-PCR and Western blot were used to detect the expressions of gene and protein of survivin.Results The recombinant adenovirus was successfully constructed,which was trasfected and duplicated in PC-3 cells perfectly.PC-3 cell proliferation was inhibited apparently and apoptosis was induced too.Ninety hours after transfection,the expressions of survivin mRNA and protein were decreased by （72.23±1.11）% and （71.75±2.64）%,respectively,and the apoptosis rate achieved to （14.43±0.57）%,which were all significanly different from those in control groups（P0.01）.Conclusion The adenovirus vector-mediated survivin siRNA can efficiently inhibit the survivin expression in PC-3 cells.The inhibition of survivin expression by a combination of RNAi and adenovirus carrier system could create a new modus to cure the androgen-independent prostate cancer.

关 键 词：前列腺癌 生存素 腺病毒 

分 类 号：R737[医药卫生—肿瘤]