Identification of miRNAs regulating pancreatic duodenal homeobox gene 1 expression during the induction of bone marrow mesenchymal stem cells differentiation into insulin-producing cells

doi:10.3969/j.issn.0529-1356.2014.02.011

Abstract

Abstract:

Objective To identify the miRNAs which may regulate the expression of pancreatic duodenal homeobox gene 1(PDX-1) during the induction of bone marrow mesenchymal stem cells(BMSCs) differentiation into insulinproducing cells. Methods BMSCs were isolated from bone marrow and induced to differentiate into insulin-producing cells using conophylline and nicotinamide. The miRNAs which may regulate the expression of PDX-1 were predicted using miRanda and TargetScan and identified by dual luciferase report system. The expressions of miRNAs and PDX-1 were determined using Real-time PCR during the induction of BMSCs differentiation into insulin-producing cells. Results The induced cells were stained scarlet by DTZ and the expression of insulin was positive by immunofluorescence cytochemistry. Four miRNAs which may regulate the expression of PDX-1 were firstly obtained by bioinformatics.Using dual luciferase reporter system in vitro, two of the four miRNAs, miR-149and miR-346, were proved to effectively inhibit PDX-1 expression, respectively, by binding to the 3’UTR of PDX-1 mRNAs. The Real-time PCR results showed that the expression of miR-149 or miR-346 was negatively correlated with that of PDX-1 mRNA. Conclusion The results indicated that miR-149 and mi-346 may negatively regulate the expression of PDX-1 during the induction of insulin-producing cells.

WANG Tao MA Yun-sheng MU Chang-zheng*. Identification of miRNAs regulating pancreatic duodenal homeobox gene 1 expression during the induction of bone marrow mesenchymal stem cells differentiation into insulin-producing cells[J]. AAS, 2014, (2): 204-210.

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