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 Hoxa1 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 Hoxa1 expression, and inhibition of miR-10a relieved the repression of Hoxa1. 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 Hoxa1 gene. Furthermore, the miR-10a accelerates endodermal differentiation, likely mediated by the up-reguation of endoderm-specific down-stream genes.