MicroRNA-7-5p regulates osteogenic differentiation of hMSCs via targeting CMKLR1

B. Chen, J. Meng, Y.-T. Zeng, Y.-X. Du, J. Zhang, Y.-M. Si, X. Yuan

Department of Orthodontics, Qingdao Municipal Hospital, Nanjing Medical University, Qingdao, China. yuanxiaoqd@163.com

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• Stem Cells

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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