miR-30a和miR-30b靶向GW182的生物学功能研究

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GW182 siRNA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。

Study of GWI82 Targeted by miR-30a and miR-30b

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.