miR­204 inhibits the osteogenic differentiation of mesenchymal stem cells by targeting bone morphogenetic protein 2.

Mesenchymal stem cells (MSCs) are used to investigate regeneration and differentiation. MicroRNA­204 (miR­204) in involved in the Runt­related transcription factor 2/alkaline phosphatase/bone morphogenic protein 2 (Runx2/ALP/BMP2) signaling pathway that regulates bone marrow mesenchymal stem cell (BMSC) differentiation; however, the mechanisms underlying the effects of miR­204 are yet to be determined. The aim of the present study was to investigate the effects of miR­204 on BMSC differentiation. BMSCs were derived from rat bone marrow. The expression levels of Runx2, ALP and BMP2 were measured via reverse transcription­quantitative polymerase chain reaction and western blot analyses following transfection of BMSCs with miR­204 agomir or BMP2 expression vector. The ability of the miR­204 gene to directly bind BMP2 mRNA was assessed using dual­luciferase assays. Ossification was measured via alizarin red stain assays. It was observed that the expression levels of Runx2 and ALP increased over time, whereas those of miR­204 decreased; additionally, miR­204 agomir upregulation inhibited the expression of Runx2, ALP and BMP2 in BMSCs. It was revealed that miR­204 directly interacted with BMP2 mRNA, and that transfection with miR­204 agomir suppressed ossification in BMSCs by targeting the BMP2/Runx2/ALP signaling pathway.