miR-193b在HepG2细胞中下调K-Ras蛋白表达  

Downregulation of K-ras expression in HepG2 cells by miR-193b

作　　者：杨永强[1] 付汉江[1] 铁轶[1] 刘珊珊[1,2] 朱捷[1] 邢瑞云[1] 郑晓飞[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所,北京100850 [2]吉林大学生命科学学院,长春130012

出　　处：《军事医学科学院院刊》2010年第3期216-219,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家重点基础研究发展计划(973计划)(2010CB912801;2007CB914601);国家自然科学基金(30873008;30870529)

摘　　要：目的探讨microRNA-193b(miR-193b)在HepG2细胞中对K-Ras蛋白表达的调控作用。方法 Tar-getScan软件预测获得miR-193b调控候选靶基因K-ras;构建包含预测miR-193b作用靶位点的K-rasmRNA3′UTR野生型和突变型荧光素酶报告基因载体pGL-KRAS和pGL-Mu-KRAS,检测转染HepG2细胞的荧光素酶活性;合成miR-193b的dsRNA,转染HepG2细胞,实时定量PCR和Western印迹检测对K-rasmRNA和蛋白表达的影响。结果与结论 miR-193b可以与K-rasmRNA3′UTR结合。在HepG2中,miR-193b在mRNA和蛋白水平下调K-ras基因表达。Objective To investigate the effect of microRNA-193b（miR-193b）on the expression of K-ras oncogene in HepG2 cells.Methods The candidate target gene K-ras of miR-193b was predicted by TargetScan software.The luciferase report vector with wild or mutant K-ras mRNA 3＇UTR was constructed respectively.The luciferase activity of HepG2 cells transfected by synthesized dsRNA of miR-193b and the luciferase report vectors were detected.The synthesized dsRNA of miR-193b was transfected into HepG2 cells,and the qRT-PCR and Western blotting were used to assay the expression of K-Ras.Results and Conclusion miR-193b can bind to the 3＇UTR of K-ras mRNA,and miR-193b downregulates the mRNA and the protein expression of K-ras in HepG2 cells.

关 键 词：微RNAS miR-193b HEPG2 K-RAS 突变 转染 聚合酶链反应 基因表达 

分 类 号：R363[医药卫生—病理学]