MicroRNA-338-3p真核表达载体的构建及其对胃癌细胞SGC-7901增殖的影响  

Construction of microRNA-338-3p overexpression vector and its effects on proliferation of gastric cancer cell SGC-7901

作　　者：郭波[1] 王文静[2] 赵正豪 赵凌宇[1] 汪鲁敏 胡丽丽[1] 宋土生[1] 黄辰[1,3] 

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 [2]西安交通大学医学院第一附属医院肝胆外科,陕西西安710061 [3]西安交通大学医学院心血管研究中心,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2014年第4期442-446,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81171398);陕西省科技攻关项目重大科技专项(No.2010ZDKG-50)~~

摘　　要：目的构建microR-338-3p(miR-338-3p)过表达载体并探究其对人胃癌SGC-7901细胞系体外增殖的影响。方法以pcDNATM6.2-GW/EmGFP-miR真核表达载体为基础,经EcoRⅠ和HindⅢ双酶切和纯化后与人工合成miR-338-3p的寡聚核苷酸进行连接,转化至E.Coli Top10后,经过DNA测序、BLAST检测及验证;将构建成功的载体瞬时转染入SGC-7901细胞系中,应用Real-time PCR检测miR-338-3p的表达水平,利用MTT法检测细胞增殖的改变。结果测序验证成功构建了miR-338-3p的真核表达载体;瞬时转染SGC-7901细胞后miR-338-3p表达水平有显著增加,能够抑制SGC-7901细胞的体外增殖。结论成功构建了miR-338-3p的真核表达载体,获得了瞬时表达miR-338-3p的人胃癌SGC-7901细胞系,初步证实miR-338-3p过表达可抑制肿瘤细胞的增殖。Objective To construct an overexpression vector of microR-338-3p （miR-338-3p） and detect its effect on the proliferation of human gastric cancer cell line SGC-7901.Methods pcDNATM6.2-GW/EmGFP-miR eukaryotic expression vector was cut using restriction site of Hind Ⅲ and EcoR Ⅰ,connected to miR-338-3p oligonucleotide,and identified by sequencing and blast.miR-338-3p overexpression vector was transfected into SGC-7901 cells and the expression level was determined by real-time PCR.The proliferation of SGC-7901 cells was also examined by MTT.Results The overexpression vector of miR-338-3p was identified as real by sequencing.The expression level of miR-338-3p in SGC-7901 cells with expression vectors transfected was increased significantly （P＜0.05）.The proliferation of SGC-7901 cells in vitro was also inhibited.Conclusion The overexpression vector of miR-338-3p was successfully constructed and human gastric cancer cell SGC-7901 instantaneously expressing miR-338-3p was obtained,which preliminarily confirmed that the overexpression of miR-338-3p can inhibit the proliferation of cancer cells.

关 键 词：miR-338-3p 胃癌 表达载体 细胞增殖 

分 类 号：R34[医药卫生—基础医学]