siRNA对HepG2细胞mcl-1表达的影响  

Effect of siRNA on mcl-1 expression in HepG2 cells

作　　者：张巍[1] 赵蒲[2] 曹大勇[1] 陶开山[1] 蔡磊[1] 张福琴[1] 窦科峰[1] 

机构地区：[1]第四军医大学西京医院肝胆外科,陕西西安710033 [2]第四军医大学基础部细胞生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2007年第6期530-533,共4页Journal of the Fourth Military Medical University

摘　　要：目的:探讨特异性siRNA对肝癌细胞系HepG2中mcl-1表达的影响.方法:将特异性siRNA经脂质体介导转染人肝癌细胞系HepG2;半定量RT-PCR观察干扰前后mcl-1 mRNA水平变化,比较对应不同位点的两对siRNA片段对mcl-1的干扰效果,分析RNA干扰的特异性、时效性;Western Blot检测转染前后Mcl-1蛋白表达情况.结果:特异性siRNA片段能有效降低mcl-1 mRNA水平,最大干扰效率达67.25%,高于作为对照的非相关片段;对应不同位点的两对siRNA片段对mcl-1均可产生干扰作用,彼此间差别不大;干扰作用于转染后24h即可出现,48h达高峰,72h稍有降低;Western Blot证明转染siRNA-F1后Mcl-1蛋白表达下调.结论:RNA干扰技术可有效下调肝癌细胞系HepG2中mcl-1 mRNA水平,明显下调Mcl-1蛋白表达.AIM： To investigate the effect of specific siRNA on racl-1 expression in hepatoma cell line HepG2. METHODS： Human hepatoma cell line HepG2 was cultured in vitro and transfected with two siRNAs （different loci ） of mcl-1 by lipofectamine 2000. The levels of mcl-1 mRNA expression before and after transfection were＂ detected by semi-quantitative reverse transcription polymerase chain reaction （ RT-PCR ）. The interfering effect between the two siRNAs and the specificity and time effect for interference were compared. Then the levels of Mcl-1 protein expression before and after transfection were detected by Western Blot. RESULTS： The level of mcl-1 mRNA in HepG2 cells was inhibited by the specific siRNAs. The expression of mcl-1 mRNA began to decrease 24 h after transfection; and the most apparent interfering efficiency was 67.25% 48 h after transfection, which was markedly higher than that in the cells transfected with the control siRNAs; at 72 h, the expression restored a little. Both siRNAs （ siRNA-F1 and siRNA-F2 ） from different loci had interfering effect on mcl-1 mRNA expression, and there was only a little difference between them. Western Blot indicated that the expression of Md-1 protein was downregulated in HepG2 cells after interfered with siRNA-F1. CONCLUSION： RNA interference can downregulate the level of mcl-1 mRNA in HepG2 cells. And Mcl-1 protein expression is also downregulated by specific siRNA.

关 键 词：癌 肝细胞 mcl—1 RNA干扰 

分 类 号：R735.7[医药卫生—肿瘤]