microRNA-1297 promotes the progression of osteoporosis through regulation of osteogenesis of bone marrow mesenchymal stem cells by targeting WNT5A

摘要

OBJECTIVE: This study was designed to investigate whether microRNA-1297 can regulate the osteogenesis of bone marrow mesenchymal stem cells (BMSCs) through WNT5A, thus influencing the progression of osteoporosis. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) assay was performed to analyze microRNA-1297 level and osteogenesis-related markers in osteoporosis patients and controls. The protein levels of the above markers and WNT5A were detected by Western blot. Alkaline phosphatase (ALP) activity assay and ALP staining were used to measure the degree of osteogenic differentiation under the control of microRNA-1297 and WNT5A, and ARS staining was used to detect the mineralization ability of hBMSC after overexpression of microRNA-1297. The binding sites of microRNA-1297 and WNT5A were determined by the dual luciferase-reporting assay. Besides, the activity of Wnt signal transduction pathway in different treatment groups was detected by TOP/FOP report. RESULTS: MicroRNA-1297 was highly expressed in osteoporotic patients, and its level decreased significantly with the increasing of osteogenic induction. Bioinformatics prediction suggested that microRNA-1297 can target WNT5A. In vitro experiments showed that overexpression of microRNA-1297 in hBMSC can reduce the level of WNT5A, while interference with microRNA-1297 can increase the level of WNT5A. Overexpression of microRNA-1297 and transfection of si-WNT5A significantly reduced the mRNA levels of RUNX2, OSX, ALP, OCN, OPN and COL1A1, thereby inhibiting osteogenic differentiation. Overexpression of microRNA-1297 could interfere with WNT signaling pathway regulation and regulate the osteogenic differentiation of hBMSCs. CONCLUSIONS: microRNA-1297 could regulate the osteogenesis of BMSCs by combining with WNT5A so as to accelerate the progression of osteoporosis.