基础医学与临床 ›› 2016, Vol. 36 ›› Issue (5): 598-603.

• 研究论文 • 上一篇    下一篇

miR-92b-3p促进人脐带间充质干细胞的成骨分化

张燕,严冰浩,徐辰,顾道兰,王越,刘厚奇   

  1. 第二军医大学
  • 收稿日期:2015-11-06 修回日期:2016-01-04 出版日期:2016-05-05 发布日期:2016-04-26
  • 通讯作者: 刘厚奇 E-mail:houqiliu@163.com
  • 基金资助:
    国家重点基础研究和开发项目;中国国家自然科学基金

miR-92b-3p promotes the osteogenesis of humans umbilical cord mesenchymal stem cells

  • Received:2015-11-06 Revised:2016-01-04 Online:2016-05-05 Published:2016-04-26
  • Supported by:
    The National Key Basic Research and Development Project;National Natural Science Foundation of China

摘要: 目的 探究miR-92b-3p在人脐带间充质干细胞 (hUCMSCs) 成骨分化过程中的作用及其作用机制。方法 通过转染hUCMSCs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hUCMSCs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hUCMSCs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hUCMSCs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hUCMSCs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hUCMSCs成骨分化。

关键词: mir-92b-3p, 人脐带间充质干细胞(hUC-MSCs), 成骨分化, DKK1

Abstract: Objective To investigate the functional role of the miR-92b-3p in osteogenesis of human umbilical cord mesenchymal stem cells (hUCMSCs). Methods The expression level of miR-92b-3p after transfection of synthetic miR-92b-3p mimic was confirmed by qRT-PCR. The transfected hUCMSCs were induced to 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 expressions. Using 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 could significantly improve the expression of DKK1. Conclusion: MiR-92b-3p can promote the osteogenesis of hUCMSCs by downregulating the DKK1.

Key words: miR-92b-3p, hUC-MSCs, Osteo-differentiation, DKK1