目的:研究miR-143调控人骨髓问充质干细胞(hMSCs)成脂分化的新机理。方法:将NC、miR-143、siPTN、miR-143i转入hMSCs中,诱导成脂分化,检测miR-143对成脂分化的影响。经miRNA靶点分析软件Findtar预测出miR-143在人多效生长因子(hPTN)的3'-UTR端有靶点。RT-PCR、westernblot研究mjR-143与hPTN的关系。构建hPTN3'-UTR靶位点荧光检测质粒prltk-PTN及其突变质粒prltk-m,验证miR-143是否在人PTN3'-UTR上有靶点。结果:miR-143促进hMSCs成脂分化,抑制hPTN的mRNA和蛋白表达水平。荧光报告实验证实miR-143在人PTN的3'-UTR上有靶点。结论:miR-143通过与hPTN3I-UTR上的靶点相结合而抑制hPTN的表达,从而促进了hMSCs成脂分化进程。
Objective: To investigate the mechanism ofmiR-143 in the regulation of hMSCs adipogenesis. Methods: hMSCs were transfected with NC/miR-143/siPTN/miR-143i and induced into adipogenic differentiation to detect the effect ofmiR-143 on adipogenesis. The miR-143 target in 3'-UTR of hPTN was predicted by Findtar-soflware of miRNA. The relationship between miR-143 and hPTN was researched by the methods of RT-PCR and western blot. The target of miR-143 predicted in hPTN 3'-UTR was verified by luciferase expression of prltk-PTN plasmid inserted with hPTN 3'-UTR segment and prltk-m plasmid of mutant target. Results: miR-143 promoted adipogenic differentiation of hMSCs, and suppressed the expression level of hPTN mRNA and protein. The experiment of luciferase demonstrated that there was miR-143 target in hPTN 3'-UTR. Conclusion: miR-143 suppressed the expression of hPTN by combining with target in hPTN 3'-UTR to accelerate the process ofhMSCs adipogenesis.