Jichu yixue yu linchuang (Aug 2023)
LncRNA FGD5-AS1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells by regulating miR-93-5p/BMP2 axis
Abstract
Objective To investigate the impact of long non-coding RNA (lncRNA) FGD5-AS1 on the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) through regulation of the microRNA miR-93-5p/bone morphogenetic protein-2(BMP2) axis. Methods hBM-MSCs in logarithmic growth phase were taken, and the expression levels of lncRNA FGD5-AS1, miR-93-5p and BMP2 mRNA were detected before and after osteogenic differentiation; The cells were transfected or co-transfected with pcDNA FGD5-AS1, miR-93-5p inhibitor, miR-93-5p mimics and corresponding negative controls, respectively, then divided into pcDNA NC group, pcDNA FGD5-AS1 group, inhibitor NC group, miR-93-5p inhibitor group, pcDNA FGD5-AS1+mimics NC group, or pcDNA FGD5-AS1+miR-93-5p mimics group and non-transfected cells were taken as blank group. CCK-8 assay was applied to detect cell proliferation ability of each group; Alkaline phosphatase (ALP) kit was applied to detect its activity; Alizarin red staining was applied to identify cellular mineralized nodule formation; Western blot was applied to detect the levels of BMP2, osteogenesis-related markers-osteocalcin (OCN), osteopontin (OPN), and osterix(OSX); Dual-luciferase experiment was applied to verify the targeting relationship of miR-93-5p with FGD5-AS1 and BMP2, respectively. Results lncRNA FGD5-AS1 and BMP2 were found to be targets of miR-93-5p. After osteogenic differentiation, the expression of FGD5-AS1 and BMP2 was increased and the expression of miR-93-5p was decreased (P＜0.05). Compared with pcDNA NC group, the expression of FGD5-AS1 in pcDNA FGD5-AS1 group was significantly increased(P＜0.05), indicating successful transfection; Mineralized nodules, cell proliferation, ALP activity and the expression of BMP2, OCN, OPN and OSX were obviously higher (P＜0.05) in pcDNA FGD5-AS1 group. Compared with the inhibitor NC group, the expression of miR-93-5p in miR-93-5p inhibitor group was significantly decreased (P＜0.05), indicating successful transfection; Mineralized nodules, cell proliferation, ALP activity and the expression of BMP2, OCN, OPN and OSX were obviously improved (P＜0.05) in miR-93-5p inhibitor group. Over-expression of miR-93-5p inhibited the promoting effect of FGD5-AS1 on the osteogenic differentiation of hBM-MSCs(P＜0.05). Conclusions Up-regulation of FGD-AS1 promotes the osteogenic differentiation of hBM-MSCs, which might be related to the miR-93-5p/BMP2 axis.
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