miRNA干扰Rap1对肝癌细胞凋亡的影响  

miRNA interference-mediated Rap1 gene silencing increases apoptosis in hepatocellular carcinoma cell line HepG2

作　　者：甘平[1] 查勇[1] 姚乾[1] 陈真[1] 谭晶[1] 

机构地区：[1]昆明医科大学第三附属医院(云南省肿瘤医院)腹部外科,云南省昆明市650118

出　　处：《世界华人消化杂志》2013年第10期894-898,共5页World Chinese Journal of Digestology

基　　金：云南省自然科学应用基础研究基金资助项目;No.2011FB064~~

摘　　要：目的:探讨抑制/激活蛋白(repressor/activator protein,Rap1)对肝癌细胞凋亡的影响及其作用机制.方法:肝癌细胞HepG2实验分组为:Rap1miRNA转染组、空质粒组、对照组(空白对照组).用流式细胞仪检测各组肝癌细胞凋亡率.采用Westernblot检测各组细胞中Rap1的表达,用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测核因子κB(nuclear factor-κB,NF-κB)p65的表达.结果:流式细胞仪检测结果提示,转染后72h,Rap1 miRNA组肝癌细胞凋亡率,比空质粒组和对照组高(33.0%±5.8%vs8.2%±2.5%,0.6%±0.2%,P<0.05).Rap1 miRNA转染组中Rap1,NF-κBp65表达下降.结论:miRNA干扰Rap1能促进肝癌细胞凋亡.同时伴有NF-κBp65表达下降.AIM： To investigate the role of repressor/activator protein 1 （Rap1） in nuclear factor-κB （NF-κB）-mediated apoptosis of liver cancer cells. METHODS： Hepatocellular carcinoma HepG2 cells were divided into three groups： control group, empty plasmid-transfected group, and Rap1 miRNA group. Apoptosis was determined by flow cytometry 72 h after transfection. The expression of Rap1 protein was measured by Western blot, and the expression of NF-κB p65 mRNA was measured by RT-PCR. RESULTS： Compared to the control group and empty plasmid-transfected group, the level of Rap1 was significantly decreased and apoptosis rate （33.0% ± 5.8% vs 8.2% ± 2.5%, 0.6% ± 0.2%, both P0.05） was significantly increased 72 h after miRNA transfection. Cells in the Rap1 miRNA group had a significant reduction in the level of NF-κB p65 mRNA compared to controls.CONCLUSION： Rap1 miRNA increases apopto- sis in hepatocellular carcinoma cell line HepG2 by decreasing the expression of NF-κB p65.

关 键 词：肝细胞肝癌 抑制 激活蛋白 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]