LncRNA XIXT promotes osteogenic differentiation of bone mesenchymal stem cells and alleviates osteoporosis progression by targeting miRNA-30a-5p

H.-L. Zhang, X.-Y. Du, Q.-R. Dong

Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China. qirong_dong@163.com

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• Stem Cells

OBJECTIVE: To uncover the role of XIXT in influencing the osteogenesis of hBMSCs by adsorbing microRNA-30a-5p (miRNA-30a-5p) to upregulate RUNX2.

PATIENTS AND METHODS: The serum samples were collected from osteoporosis patients and normal people. hBMSCs were isolated from femoral head tissues. The serum levels of XIXT and miRNA-30a-5p were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The expression levels and activities of the osteogenic differentiation-related genes in hBMSCs after transfection of sh-XIXT, sh-RUNX2, miRNA-30a-5p mimic, and inhibitor were detected by qRT-PCR, Western blot, ALP activity assay, and alizarin red staining. The Dual-Luciferase Reporter Gene Assay was performed to confirm the binding of XIXT to miRNA-30a-5p, as well as the binding of miRNA-30a-5p to RUNX2.

RESULTS: LncRNA XIXT was significantly downregulated, and miRNA-30a-5p was upregulated in the serum of osteoporosis patients. The osteogenic differentiation-related genes (ALP, RUNX2) and XIXT were markedly upregulated in a time-dependent manner, while miRNA-30a-5p level gradually decreased in hBMSCs with the prolongation of osteogenesis. The knockdown of XIXT inhibited the osteogenic differentiation of hBMSCs. In hBMSCs, XIXT regulated RUNX2 expression by targeting miRNA-30a-5p. The knockdown of miRNA-30a-5p partially reversed the inhibitory effect of XIXT on the osteogenesis of hBMSCs. However, the downregulated RUNX2 reversed the promotive effect of miRNA-30a-5p on the osteogenesis of hBMSCs.

CONCLUSIONS: LncRNA XIXT upregulated RUNX2 by absorbing miRNA-30a-5p, and thus induced hBMSCs osteogenesis to alleviate osteoporosis.

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