摘要
目的研究microRNA-10a(miR-10a)参与的转录后调控,提高人胎盘间充质干细胞(PMSC)向内胚层细胞分化的能力。方法利用慢病毒过表达miR-10a前体和反义寡核苷酸抑制miR-10a表达,分析miR-10a与其调控的靶基因Hoxa1的表达关系;并采用实时定量PCR(qRT-PCR)和免疫荧光细胞化学染色检测miR-10a过表达后,PMSC向内胚层细胞分化能力的变化。结果 PMSC中过表达和抑制表达miR-10a,能反向调控其靶基因Hoxa1的表达;miR-10a过表达后,内胚层细胞特异性基因FoxA2、Sox-17、Pdx-1和Cdx2的表达量显著上升,FOXA2、SOX-17和PDX-1阳性细胞数量明显增多。结论 miR-10a在化学诱导向内胚层细胞分化的PMSC中呈现高表达;miR-10a可能通过抑制其靶基因Hoxa1的表达,调控下游内胚层基因表达上调,从而促进胎盘间充质干细胞向内胚层细胞分化。
Objective To improve the potential of endodermal differentiation of human placenta-derived mesenchymal stem cells （PMSCs） by microRNA-10a （miR-10a）-mediated post-transcriptional regulation of its mRNA targets. Methods Lentiviral vectors were used to stably and specifically over-express miR-10a and inhibited the miR-10a function by its antagomir. In addition, the relationship between miR-10a and Hoxal expression was analyzed. Real-time quantitative PCR （qRT-PCR） and immunofluorescent cytochemical staining were utilized to test the mRNA and protein expression variation and assess the ability of PMSCs to differentiate into endodermal cells. Results Over-expression of miR-10a led to the suppression of endogenous Hoxal expression, and inhibition of miR-10a relieved the repression of Hoxal. Over-expression of miR-10a in PMSCs resulted in the up-regulation of endoderm-specific genes （FoxA2, Sox-17, Pdx-1 and Cdx2） and the increased proportions of FOXA2, SOX-17 and PDX-1 positive events as compared with the control treated cells. Conclusion miR-10a was up-regulated during endodermal differentiation of PMSCs and involved in its differentiation partially via the suppression of the Hoxal gene. Furthermore, the miR-10a accelerates endodermal differentiation, likely mediated by the up-reguation of endoderm-specific down-stream genes.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2013年第10期1015-1019,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
宁夏医科大学科研项目(XQ2011023,XQ2012003)
宁夏自然科学基金(NZ1212)
