Identification Of Microrna Regulating PDX1Expression During The Induction Of Bmscs Differentiation Into Insulin-Producing Cells

ObjectiveTo identify the miRNAs which may regulate the expression ofPDX1during the induction of bone marrow mesenchymal stemcells(bMSCs) differentiation into insulin-producing cells.MethodsBMSCs were isolated from bone marrow and induced todifferentiate into insulin-producing cells using conophylline andnicotinamide. The miRNAs which may regulate the expression ofPDX1were predicted using miRanda and TargetScan and identified bydual luciferase report system. The expressions of miRNAs and PDX1were determined using qRT-PCR during the induction of bMSCsdifferentiation into insulin-producing cells.ResultsThe induced cells could be stained scarlet by DTZ and theexpression of insulin was positive by immunofluorescencecytochemistry. Four miRNAs which may regulate the expression ofPDX1were firstly obtained by bioinformatics.Using dual luciferasereporter system in vitro, two of the four miRNAs, miR-149and miR-346,were proved to effectively inhibit PDX1expression, respectively, bybinding to the3′UTR of PDX1mRNAs. The qRT-PCR results showedthat the expression of miR-149or miR-346is negatively correlatedwith that of PDX1mRNA. ConclusionsThese results indicated that miR-149and miR-346may negativelyregulate the expression of PDX1during the induction ofinsulin-producing cells.