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

Ming Wu; Hongyan Wang; Dece Kong; Jin Shao; Chao Song; Tieyi Yang; Yan Zhang

doi:10.7717/peerj.12228

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

PeerJ. 2021 Sep 28:9:e12228. doi: 10.7717/peerj.12228. eCollection 2021.

Authors

Ming Wu^#¹, Hongyan Wang^#², Dece Kong², Jin Shao², Chao Song², Tieyi Yang², Yan Zhang²

Affiliations

¹ Postgraduate Training Base in Shanghai Gongli Hospital, Ningxia Medical University, Shanghai, China.
² Department of Orthopaedics, Gongli Hospital of Pudong New Area, Shanghai, China.

^# Contributed equally.

Abstract

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.

Keywords: MC3T3-E1; Osteoblast differentiation; Osteoporosis; Smad4; miR-452-3p.

Grants and funding

The present study was supported by the National Natural Science Foundation of China (Grant No. 81772383) and funded by Outstanding Leaders Training Program of Pudong Health Bureau of Shanghai (Grant No. PWRL2019-01). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.