Effect of the miR-186 target-regulating the osteopontin expression on the osteogenic differentiation of bone marrow mesenchymal stem cells

Objective: To explore the effects of the miR-186 target-regulating the osteopontin(OPN) expression on the osteogenic differentiation of bone marrow mesenchymal stem cells(bMSCs).Methods: The bMSCs were isolated from bone marrow,induced with BMP 2,and identified using the Alkaline Phosphatase(ALP) and Von-kossa staining.Targeting pairing relationship between miR-186 and OPN was predicted by bioinformatics,and identified with luciferase assay.After transfecting miR-186 mimics and siRNA interfering OPN gene into cells,the expressions of miR-186 and OPN were determined by real-time PCR,and the protein expression of OPN was detected using western blot.Results: The uniform distribution of bMSCs was found under light microscope,and the induced cells could be stained blue by ALP.MiRanda and TargetScan showed that miR-186 was well complementary with OPN gene.The result of luciferase assay showed that miR-186 could inhibit the expression of OPN mRNA.The results of real-time PCR and western blot showed that the over-expression of miR-186 could downregulate the expressions of OPN mRNA and protein.Conclusions: MiR-186 can inhibit the osteogenic differentiation of bMSCs by downregulating the OPN gene expression.