miR-195表达载体构建及其对肝癌细胞株HepG2植瘤抑制效果的影响  

Construction of miR-195 expression vector and anti-tumorigenic effect of miR-195 on human hepatoma cell strain HepG2

作　　者：胡晓岩[1] 姚嘉宜[1] 李梦鹤[1] 倪磊[1] 王爱英[1] 黄辰[1] 宋土生[1] 

机构地区：[1]西安交通大学医学院遗传学与分子生物学系,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2013年第1期11-15,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省科技攻关项目重大科技专项(No.2010ZDKG-50)~~

摘　　要：目的研究miR-195对人肝癌细胞株HepG2肿瘤形成的影响。方法应用分子克隆技术构建miR-195表达载体;用脂质体转染以及克隆筛选技术,建立miR-195高表达HepG2细胞株,并通过测序及序列比对等进行鉴定;建立荷瘤小鼠模型,检测不同细胞系的肿瘤生长能力。结果通过测序及序列比对,证明miR-195表达载体构建成功;荧光显微镜观察结果显示,miR-195高表达HepG2细胞株成功构建,实时定量PCR鉴定结果表明,构建的miR-195高表达HepG2细胞株miR-195表达量为对照培养HepG2细胞株的2 000倍(P<0.01);植瘤实验证明,miR-195高表达可以抑制肿瘤的形成过程。结论作为抑癌的miRNA,miR-195有望成为肝癌基因治疗的效应分子。Objective To investigate the anti-tumorigenic effect of miR-195 on human hepatoma cell strain HepG2. Methods We constructed miR-195 expression vector by molecular cloning technique, transfected it into HepG2 cell strain by liposome, and then constructed miR-195 overcxpression HepG2 cell strain by clone selection. Furthermore, we detected the tumorigenic potential of each different cell strain in vivo in nude mice. Results We successfully constructed miR-195 overexpression vector by analyzing both of the sequencing results and the fluorescent results. Furthermore, real time PCR results revealed that miR-195 expression level in miR-195 overexpression HepG2 cell strain was 2 000 as high as that of the related non-transfection HepG2 cells （P〈0.01）. Moreover, miR-195 overexpression cells exhibited less tumorigenic potential than that of the non-transfected groups. Conclusion miR-195 may act as a tumor suppressor in hepatoma development, suggesting an effective target in hepatoma therapy.

关 键 词：miR-195 HEPG2细胞株 动物模型 分子克隆技术 转染 肝癌 基因治疗 

分 类 号：Q78[生物学—分子生物学]