摘要
目的探究miR-92b-3p在人脐带间充质干细胞(h UCMSCs)成骨分化过程中的作用及其作用机制。方法通过转染h UCMSCs过表达或抑制miR-92b-3p,qRT-PCR检测各组成骨相关基因OCN、RUNX2、OSTERIX mRNA表达以明确miR-92b-3p对h UCMSCs成骨分化的体外调控作用,用茜素红染色比较各处理组的骨化能力。通过生物信息学分析miR-92b-3p可能的靶基因,并通过双荧光素酶基因报告系统以及Western blot验证。结果 miR-92b-3p在h UCMSCs成骨过程中表达量较对照组明显升高(P<0.05);过表达miR-92b-3p能促进h UCMSCs体外成骨分化及体内的异位成骨能力(P<0.05);而抑制miR-92b-3p则能降低h UCMSCs体外成骨分化(P<0.05)。过表达miR-92b-3p能显著降低DKK1的表达量(P<0.05),抑制则能明显提高DKK1的表达(P<0.05)。结论 miR-92b-3p可通过抑制DKK1表达,促进h UCMSCs成骨分化。
Objective enchymal stem cells To investigate the function of the miR-92b-3p in osteogenesis of human umbilical cord mes- （hUCMSCs）. Methods The expression of miR-92b-3p after transfection of synthetic miR- 92b-3p mimic was examined by qRT-PCR. The transfected hUCMSCs were induced into osteoblast differentiation. Alizarin red S staining was used to determine osteogenic ability of different transfected cells, qRT-PCR was used to analyze the osteogenic marker gene expression. We used bioinformatics analysis to predict the potential target of miR-92b-3p, and the dual luciferase report gene system and Western blot to verify the predication. Results The expression of miR-92b-3p in the process of osteodifferentiation was significantly higher than that of the control group （P 〈 0.01 ）. Overexpressed miR-92b-3p could promote hUCMSCs osteogenetic ability. While inhibiting miR-92b- 3p could reduce the osteogenetic differentiation level of hUCMSCs. Overexpressed miR-92b-3p could significantly reduce the expression of DKK1, while inhibition of miR-92b-3p may significantly improve the expression of DKK1. Conclusions miR-92b-3p can promote the osteogenesis of hUCMSCs by downregulating the DKK1.
出处
《基础医学与临床》
CSCD
2016年第5期598-603,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(31471390)
国家重点基础研究和开发项目(2011CB965101)
