[No authors listed]
Mesenchymal stem cells (MSCs) are used to investigate regeneration and differentiation. MicroRNAâ204 (miRâ204) in involved in the Runtârelated 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 miRâ204 are yet to be determined. The aim of the present study was to investigate the effects of miRâ204 on BMSC differentiation. BMSCs were derived from rat bone marrow. The expression levels of Runx2, ALP and BMP2 were measured via reverse transcriptionâquantitative polymerase chain reaction and western blot analyses following transfection of BMSCs with miRâ204 agomir or BMP2 expression vector. The ability of the miRâ204 gene to directly bind BMP2 mRNA was assessed using dualâluciferase 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 miRâ204 decreased; additionally, miRâ204 agomir upregulation inhibited the expression of Runx2, ALP and BMP2 in BMSCs. It was revealed that miRâ204 directly interacted with BMP2 mRNA, and that transfection with miRâ204 agomir suppressed ossification in BMSCs by targeting the BMP2/Runx2/ALP signaling pathway.
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