MicroRNA-200a-3p accelerates the progression of osteoporosis by targeting glutaminase to inhibit osteogenic differentiation of bone marrow mesenchymal stem cells.
Department of Orthopaedics, The Third Affiliated Hospital of Chongqing Medical University(Gener Hospital), Chongqing, 401120, China. Electronic address: [Email]
To uncover the role of microRNA-200a-3p in regulating osteogenic differentiation of MSCs via targeting glutaminase, thus influencing the progression of OP. Serum level of microRNA-200a-3p in OP patients and healthy controls was determined by qRT-PCR. MicroRNA-200a-3p level in MSCs undergoing osteogenic differentiation for different days was examined as well. ALP activity, calcification nodules and relative levels of Bglap, Runx2 and OPN in MSCs overexpressing microRNA-200a-3p undergoing osteogenic differentiation were detected. Relative l-glutaminase uptake in MSCs undergoing osteogenic differentiation for different days was determined. After transfection of si-GLS in MSCs undergoing osteogenic differentiation, l-glutaminase uptake, ALP activity and relative levels of Bglap, Runx2 and OPN were detected. The potential binding relationship between microRNA-200a-3p and GLS was tested by dual-luciferase reporter gene assay. Finally, rescue experiments were conducted to elucidate the role of microRNA-200a-3p/GLS in osteogenic differentiation of MSCs. MicroRNA-200a-3p level was higher in serum of OP patients relative to controls. Its level in MSCs gradually decreased with the prolongation of osteogenic differentiation. Overexpression of microRNA-200a-3p reduced cell viability, ALP activity, number and volume of calcification nodule. The mRNA levels of Bglap, Runx2 and OPN were downregulated by overexpressed microRNA-200a-3p. The cell viability, ALP activity, number and volume of calcification nodule were reduced when microRNA-200a-3p was knocked down. The mRNA levels of Bglap, Runx2 and OPN were upregulated when transfected microRNA-200a-3p inhibitor. l-glutaminase uptake increased with the prolongation of osteogenic differentiation in MSCs. Knockdown of GLS attenuated l-glutaminase uptake and ALP activity, as well as downregulated Bglap, Runx2 and OPN. Besides, GLS was verified to directly bind to microRNA-200a-3p. GLS overexpression reversed the inhibitory effects of overexpressed microRNA-200a-3p on osteogenic differentiation of MSCs. MicroRNA-200a-3p suppresses osteogenic differentiation of MSCs via targeting glutaminase, thereafter accelerating the progression of OP.
