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目的:研究miR-143调控人骨髓间充质干细胞(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、western blot研究miR-143与hPTN的关系。构建hPTN 3’-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PTN 3’-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hPTN3’-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。
Aim: To study the new mechanism of miR-143 regulating adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). Methods: NC, miR-143, siPTN and miR-143i were transfected into hMSCs to induce adipogenic differentiation and to detect the effect of miR-143 on adipogenic differentiation. Findtar, a miRNA target analysis software, has predicted that miR-143 has a target at the 3’-UTR of human pleiotropic growth factor (hPTN). The relationship between miR-143 and hPTN was analyzed by RT-PCR and western blot. The hPTN 3’-UTR target site fluorescence detection plasmid prltk-PTN and its mutant plasmid prltk-m were constructed to verify whether miR-143 has a target on human PTN 3’-UTR. Results: miR-143 promoted adipogenic differentiation of hMSCs and inhibited hPTN mRNA and protein expression. Fluorescent reporter experiments confirmed that miR-143 has a target on the 3’-UTR of human PTN. Conclusion: miR-143 inhibits the expression of hPTN by binding to the target of hPTN3’-UTR, which promotes the adipogenic differentiation of hMSCs.