Clinical Interventions in Aging (Jan 2021)

MicroRNA-197-3p Inhibits the Osteogenic Differentiation in Osteoporosis by Down-Regulating KLF 10

  • You M,
  • Zhang L,
  • Zhang X,
  • Fu Y,
  • Dong X

Journal volume & issue
Vol. Volume 16
pp. 107 – 117

Abstract

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Murong You, Liang Zhang, Xiaoxiang Zhang, Yang Fu, Xieping Dong Department of Orthopedics, Jiangxi Provincial People’s Hospital, Nanchang, Jiangxi Province 330006, People’s Republic of ChinaCorrespondence: Murong YouDepartment of Orthopedics, Jiangxi Provincial People’s Hospital, Nanchang, Jiangxi Province 330006, People’s Republic of ChinaEmail [email protected]: Studies have shown that microRNA (miRNA) regulates gene expression of osteoporosis (OS). It is known that miR-197-3p is abnormally expressed in osteoporosis. This study is to investigate the mechanism of miR-197-3p in regulating osteoblast differentiation.Methods: Rats were ovariectomized to establish an animal model of postmenopausal osteoporosis. The expression of miR-197-3p and KLF10 was detected in ovariectomized rat models. Primary osteoblasts and MC3T-E1 cells were divided into the control group, miR-197-3p inhibitor group, NC inhibitor group and miR-197-3p inhibitor + si-KLF10 group. The expression of miR-197-3p and Kruppel-like factor 10 (KLF10) was detected by qRT-PCR and Western blot. The relationship between miR-197-3p and KLF10 was analyzed by bioinformatics and luciferase reporter assay. Cell viability was evaluated by MTT assay. The ALP activity measurement and mineralization analysis were performed.Results: The expression of miR-197-3p was significantly raised in ovariectomized osteoporosis rats. During the differentiation of osteoblasts, the expression of miR-197-3p was significantly decreased, while the expression of KLF10 was significantly raised in primary osteoblasts and MC3E3T1 cells. The expression of RUNX2, ALP, OCN and OSX in miR-197-3p inhibitor group and MC3T3-E1 group was significantly raised, and the cell survival rate and mineralized nodule were raised as well. KLF10 may be the downstream target gene of miR-197-3p. After co-transfection of miR-197-3p inhibitor and si-klf10, ALP, Runx2, OCN and OSX mRNA, cell survival rate and mineralized nodule were significantly decreased in primary osteoblasts and MC3T3-E1 cells.Conclusion: MiR-197-3p Inhibition promoted osteoblast differentiation and reduced OS by up-regulating KLF10.Keywords: osteoporosis, miR-197-3p, KLF10, osteogenic differentiation, MC3T3-E1 cells

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