miR-30a和miR-30b靶向GW182的生物学功能研究  

Study of GW182 Targeted by miR-30a and miR-30b

作　　者：苏雪婷[1] 夏伟[1] 陈颖[1] 李少华[1] 丁红梅[1] 李慧[1] 黄皑雪[1] 赵强[1] 李洁[1] 邵宁生[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《生物技术通讯》2013年第5期604-609,共6页Letters in Biotechnology

基　　金：国家自然科学基金(31100569;30971630);国家重点基础研究发展计划(2011CB811300;2010CB912801)

摘　　要：目的:通过证明miR-30a、miR-30b靶向GW182,探索HeLa细胞中miR-30a、miR-30b及GW182的生物学功能。方法:生物信息学预测分析表明GW182的3'UTR区存在4个保守的miR-30a、miR-30b靶位点,将含有靶位点的序列片段构建到萤光素酶报告载体中,检测miR-30a、miR-30b与靶位点的结合情况;用阳离子脂质体转染GW182siRNA和miR-30a、miR-30b mimics,检测GW182下游功能的变化。结果:miR-30a、miR-30b能够靶向GW182;qPCR及Western印迹证明miR-30a、miR-30b可以在mRNA与蛋白水平上降低GW182的表达,同时影响GW182所参与的miRNA/siRNA对靶基因的抑制作用。结论:GW182是miR-30a、miR-30b的靶基因,揭示了miR-30a、miR-30b通过靶向GW182影响miRNA/siRNA作用途径的新功能。Objective： To investigate the biological function of GW182 which was a novel target of miR-30a and miR-30b in human HeLa cells. Methods： The potential target sites of miR-30a and miR-30b in the 3＇UTR of GW182 gene sequence were analyzed by bioinformatic method. Luciferase assay, quantitative real-time polymerase chain reaction and Western blot were used to confirm the target sites of miR-30a and miR-30b in GW182. Re-sults： We found that miR-30a and miR-30b repressed GW182 expression on both of the mRNA and protein lev-els. And results showed that miR-30a and miR-30b could down-regulate the effect of miR-200b targeting ZEB1 and GFP siRNA silencing the GFP expression by targeting GW182 in HeLa cell. Conclusion： These data suggest-ed that miR-30a and miR-30b may play important roles in the regulation of microRNA or siRNA function by tar-geting GW182.

关 键 词：miR-30a miR-30b GW182 靶点 

分 类 号：Q78[生物学—分子生物学]