siRNA沉默Bmi-1基因对人肝癌细胞株MHCC97-H侵袭迁移能力的影响  

SiRNA-mediated silencing of the Bmi-1gene inhibits cell invasion and metastasis in human hepatocellular carcinoma cell line MHCC97-H

作　　者：李小磊[1] 宋文杰[1] 陈战[1] 陈亚峰[1] 李霄[1] 周亮[1] 张福琴[1] 窦科峰[1] 

机构地区：[1]中国人民解放军第四军医大学西京医院肝胆胰脾外科,陕西省西安市710032

出　　处：《世界华人消化杂志》2011年第36期3643-3648,共6页World Chinese Journal of Digestology

摘　　要：目的:探讨RNA干扰技术沉默Bmi-1基因表达后,对人肝癌细胞株MHCC97-H侵袭迁移能力的影响.方法:设计并合成针对Bmi-1基因序列特异性的双链小干扰RNA(Bmi-1-siRNA),转染高转移性人肝癌细胞株MHCC97-H,用流式细胞仪观察转染效率,荧光实时定量PCR和Westernblot检测Bmi-1基因的mRNA和蛋白表达水平;通过体外Transwell小室基质侵袭和迁移实验,观察Bmi-1表达沉默后对人肝癌细胞株MHCC97-H侵袭和迁移能力的影响.结果:将针对Bmi-1基因序列特异性的小干扰RNA(Bmi-1-siRNA)转染高转移性人肝癌细胞株MHCC97-H后,流式细胞仪显示,转染效率可达到91%.与空白组、对照siRNA组相比,实验组Bmi-1-siRNA能有效抑制MHCC97-H细胞中Bmi-1基因的mRNA(F=56.199,P<0.05)和蛋白表达水平.通过Transwell小室基质侵袭和迁移实验,我们分析了不同组细胞的侵袭迁移能力.结果发现,与空白组、对照siRNA组相比,Bmi-1-siRNA转染的MHCC97-H细胞穿透能力明显降低(F=186.66,12.746,P<0.05).结论:沉默Bmi-1基因表达可抑制肝癌细胞株MHCC97-H的侵袭迁移力.AIM：To investigate the effect of small interference RNA （siRNA）-mediated silencing of the Bmi-1gene on cell invasion and metastasis in human hepatocellular carcinoma cell line MHCC97-H.METHODS：Chemically synthesized siRNA duplex targeting the Bmi-1 gene （Bmi-1-siRNA） was transiently transfected into MHCC97-H cells,which have high metastatic potential,using Lipofectamine 2000.Transfection efficiency was evaluated by flow cytometry （FCM）.Bmi-1 mRNA and protein expression was detected by real-time RT-PCR and Western blot,respectively.The effect of Bmi-1 knockdown on cell invasion and migration was analyzed by Transwell chamber assays.RESULTS：The transfection efficiency achieved using Bmi-1-siRNA was 91%.Compared to the blank group and control siRNA group,transfection with Bmi-1-siRNA effectively down-regulated the expression of Bmi-1mRNA （F=56.199,P0.05） and protein.Bmi-1-siRNA-transfected MHCC97-H cells had lower levels of invasion and migration capacity than cells in the blank group and control-siRNA group （F=186.66,12.746,both P0.05）.CONCLUSION：SiRNA-mediated silencing of the Bmi-1 gene could significantly inhibit cell invasion and metastasis in human hepatocellular carcinoma cell line MHCC97-H.

关 键 词：RNA干扰 肝肿瘤 BMI-1 侵袭 转移 

分 类 号：R735.7[医药卫生—肿瘤]