miR-532-3p inhibits osteogenic differentiation in MC3T3-E1 cells by downregulating ETS1

• Published in: Biochemical and biophysical research communications Vol. 525; no. 2; pp. 498 - 504
• Main Authors: Fan, Qingxin, Li, Yuqiao, Sun, Qiang, Jia, Yutao, He, Chao, Sun, Tianwei
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 30.04.2020
• Subjects: ETS1; MC3T3-E1; MiR-532-3p; Osteoblast; Osteoporosis; Osteoblast; Osteoporosis; MiR-532-3p; ETS1; MC3T3-E1
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2020.02.126

Many studies had identified that MicroRNAs (miRNAs) could affect bone metabolism by regulating the expression of various proteins. This study explored the effect and mechanism of miR-532-3p on osteogenic differentiation. We analyzed the content of miR-532-3p in osteoporosis patients, osteoporosis rats, and osteogenic induced MC3T3-E1 cells. MiR-532-3p mimic or inhibitor utilized to alter intracellular miR-532-3p content. MTT method executed to detect the effect of miR-532-3p on osteoblast proliferation. Real-time qPCR, Western blot, alkaline phosphatase staining, and alizarin red staining utilized to ascertain the influence of miR-532-3p on osteogenic differentiation. Then, databases and a dual-luciferase reporter gene assay used to verify the target of miR-532-3p. Furthermore, the lentiviral vector was utilized to overexpress interesting target gene expression and checked whether the target gene was involved in the regulation of osteogenic differentiation by miR-532-3p. MiR-532-3p expression boosted in low bone mineral density (BMD) patients and rats. In MC3T3-E1 cells, miR-532-3p expression gradually decreased as osteogenic induction matures. MiR-532-3p mimic negatively regulated succinate dehydrogenase (SDH) activity, alkaline phosphatase (ALP) activity, mineralization ability, the osteogenic-associated gene (Col1A1, Runx2, ALP, OPN, and OCN) and E−26 transformation specific-1 (ETS1) expression of MC3T3-E1 cells. Things are the opposite of the miR-532-3p inhibitor. ETS1 identified as the miR-532-3p target gene, and miR-532-3p could inhibit its expression. Besides, improved ETS1 expression could rescue the suppressive effect of miR-532-3p mimic on osteogenic differentiation. miR-532-3p can suppress osteogenic differentiation by downregulating ETS1 expression. •miR-532-3p is negatively correlated with bone mineral density, and ETS1 is the opposite.•miR-532-3p inhibits osteogenic differentiation of MC3T3-L1.•ETS1 prompts osteogenic differentiation of MC3T3-L1.•miR-532-3p inhibits the osteogenic differentiation of MC3T3-L1 by down-regulating ETS1.