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  

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

作　　者：陈冬梅[1] 马海滨[1] 刘淑丹[1] 王立斌[1] 李玉奎[1] 魏军[1] 

机构地区：[1]宁夏医科大学总医院干细胞研究所生育力保持教育部重点实验室,宁夏银川750004

出　　处：《基础医学与临床》2014年第10期1309-1314,共6页Basic and Clinical Medicine

基　　金：宁夏自然科学基金(NZ13162)

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-βR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。Objective 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 measure the mR-NA and protein expression variation and to 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 pancreatic-specific genes （Ngn3,Nkx6.1,PDX-1 and Insulin） （P ＜ 0.05） and increased proportions of Insulin and PDX-1 positive events as compared to control treated cells.Conclusions 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.

关 键 词：miR-124a 转化生长因子βⅠ型受体 胰腺祖细胞 胎盘间充质干细胞 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]