miR-452-3p inhibited osteoblast differentiation by targeting Smad4

• DOI: 10.7717/peerj.12228
• Publication Date: 2021-09-28T09:03:45Z
• Authors: Ming Wu; Hongyan Wang; Dece Kong; Jin Shao; Chao Song; Tieyi Yang; Yan Zhang
• Keywords: miR-452-3p; 0301 basic medicine; 03 medical and health sciences; Osteoblast differentiation; QH301-705.5; R; Osteoporosis; Medicine; Biology (General); Smad4; MC3T3-E1; Biochemistry

Osteoblast differentiation is a complex process that is essential for normal bone formation. A growing number of studies have shown that microRNAs (miRNAs) are key regulators in a variety of physiological and pathological processes, including osteogenesis. In this study, BMP2 was used to induce MC3T3-E1 cells to construct osteoblast differentiation cell model. Then, we investigated the effect of miR-452-3p on osteoblast differentiation and the related molecular mechanism by RT-PCR analysis, Western blot analysis, ALP activity, and Alizarin Red Staining. We found that miR-452-3p was significantly downregulated in osteoblast differentiation. Overexpression miR-452-3p (miR-452-3p mimic) significantly inhibited the expression of osteoblast marker genes RUNX2, osteopontin (OPN), and collagen type 1 a1 chain (Col1A1), and decreased the number of calcium nodules and ALP activity. In contrast, knockdown miR-452-3p (miR-452-3p inhibitor) produced the opposite effect. In terms of mechanism, we found that Smad4 may be the target of miR-452-3p, and knockdown Smad4 (si-Smad4) partially inhibited the osteoblast differentiation enhanced by miR-452-3p. Our results suggested that miR-452-3p plays an important role in osteoblast differentiation by targeting Smad4. Therefore, miR-452-3p is expected to be used in the treatment of bone formation and regeneration.