OBJECTIVE: To investigate the function of microRNA-7-5p in human mesenchymal stem cells (hMSCs) and its underlying potential mechanism in osteogenic differentiation.
PATIENTS AND METHODS: Expression levels of osteogenic genes (ALP, RUNX2), microRNA-7-5p and CMKLR1 in hMSCs were detected by quantitative real time-polymerase chain reaction (qRT-PCR). After transfection of microRNA-7-5p mimics or inhibitor, the effect of microRNA-7-5p on osteogenic differentiation of hMSCs was detected by Alizarin red staining, ALP activity determination and Western blot, respectively. The potential target gene of microRNA-7-5p was predicted online and further verified by luciferase reporter gene assay. Rescue experiments were conducted to explore whether the effect of microRNA-7-5p on osteogenic differentiation of hMSCs could be reversed by CMKLR1.
RESULTS: Expression levels of ALP, RUNX2 and microRNA-7-5p were gradually elevated with the prolongation of osteogenic differentiation, whereas CMKLR1 was reduced. Overexpression of microRNA-7-5p increased levels of ALP and RUNX2. The amount of calcified nodules was increased after microRNA-7-5p overexpression. CMKLR1 was the target gene of microRNA-7-5p. The effect of microRNA-7-5p on osteogenic differentiation of hMSCs could be reversed by CMKLR1.
CONCLUSIONS: MicroRNA-7-5p promotes osteogenic differentiation of hMSCs via targeting CMKLR1.
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