基础医学与临床 ›› 2014, Vol. 34 ›› Issue (10): 1309-1314.

• 研究论文 •    下一篇

miR-124a通过抑制TGF-β通路促进人胎盘间充质干细胞向胰腺祖细胞分化

陈冬梅1,马海滨1,刘淑丹1,王立斌2,李玉奎3,魏军1   

  1. 1. 宁夏医科大学总医院
    2. 宁夏医科大学附属医院
    3. 宁夏医科大学
  • 收稿日期:2014-02-18 修回日期:2014-04-21 出版日期:2014-10-05 发布日期:2014-09-25
  • 通讯作者: 魏军 E-mail:lydiajunwei@hotmail.com
  • 基金资助:
    宁夏自然科学基金

MiR-124a accelerates pancreatic progenitor cells differentiation of mesenchymal stem cells from human placental through TGF-β pathway

  • Received:2014-02-18 Revised:2014-04-21 Online:2014-10-05 Published:2014-09-25
  • Supported by:
    The natural science foundation of ningxia

摘要: 目的 研究miR-124a参与抑制TGFBR1和BMPR1B的转录后调控,及其对人胎盘间充质干细胞(PMSCs)向胰腺祖细胞分化的影响。方法 用慢病毒过表达miR-124a前体,建立PMSCs中过表达miR-124a细胞模型;转染miR-124a反义寡核苷酸,抑制miR-124a表达;Real-time PCR分析miR-124a与其调控的靶基因TGFBR1和BMPR1B的表达调控关系;Real-time PCR、Western blot和免疫荧光化学染色检测miR-124a过表达后,胰腺祖细胞特征基因和蛋白表达变化。结果PMSCs中过表达和抑制表达miR-124a,能反向调控其靶基因TGFBR1和BMPR1B的表达;miR-124a过表达后,胰腺祖细胞特异性基因PDX-1、Nkx6.1、NGN3和β细胞特征基因Insulin的表达量显著上升(P<0.05),PDX-1和Insulin的蛋白表达水平也同样升高。结论 miR-124a可能通过抑制TGFBR1的表达,阻断TGF-β/TGF-β R信号,使细胞发生间质上皮转化,并可能通过抑制BMPR1B的表达阻碍细胞向成骨方向分化,从而指导胎盘间充质干细胞向胰腺祖细胞分化。

关键词: miR-124a,转化生长因子β I型受体,胰腺前体细胞,胎盘间充质干细胞,分化

Abstract: Objective The aim of this study is to improve the potential of pancreatic progenitor cells differentiation of human placenta derived mesenchymal stem cells (PMSCs) by miR-124a mediates post-transcriptional regulation of its mRNA targets TGFBR1 and BMPR1B . Methods We used lentiviral vectors to stably and specifically over express miR-124a and inhibited the miR-124a function by its antagomir. In addition, we analyzed the relationship between miR-124a and target genes expression. Real-time PCR, Western blot and immunofluorescent staining were used to test the mRNA and protein expression variation and assess the ability of PMSCs to differentiate into pancreatic progenitor cells. Results Overexpression of miR-124a led to repression of endogenous TGFBR1 and BMPR1B, and inhibition of miR-124a relieved the repression of TGFBR1 and BMPR1B. Overexpression of miR-124a in PMSCs resulted in up-regulated of pancreatic-specific genes (Ngn3, Nkx6.1, PDX-1 and Insulin) (P<0.05)and increased proportions of Insulin and PDX-1 positive events compared to control treated cells. Conclusion miR-124a is involved in pancreatic progenitor cells differentiation of PMSCs partially via suppression of the TGFBR1 and BMPR1B genes. Furthermore, the miR-124a accelerates pancreatic progenitor cells differentiation, possibly mediated by TGF β pathway.

Key words: miR-124a,TGF β R1,pancreatic precursor cell,placenta mesenchymal stem cells,differentiation

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