miR-210-3p suppresses osteogenic differentiation of MC3T3-E1 by targeting brain derived neurotrophic factor (BDNF)

Li Deng; Shuang Lai; Liyuan Fan; Xinlun Li; Hao Huang; Yandong Mu

doi:10.1186/s13018-022-03315-x

miR-210-3p suppresses osteogenic differentiation of MC3T3-E1 by targeting brain derived neurotrophic factor (BDNF)

J Orthop Surg Res. 2022 Sep 14;17(1):418. doi: 10.1186/s13018-022-03315-x.

Authors

Li Deng^{1

2}, Shuang Lai¹, Liyuan Fan¹, Xinlun Li³, Hao Huang⁴, Yandong Mu⁵

Affiliations

¹ Stomatology Department, Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, 611731, China.
² Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital, The Second Clinical College of North Sichuan Medical College, Nanchong, 637000, Sichuan, China.
³ Stomatology Department, Sichuan Provincial People's Hospital, Chengdu, 610072, Sichuan, China.
⁴ Key Laboratory of Advanced Technologies of Materials (Ministry of Education), School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu, 610031, Sichuan, China.
⁵ Stomatology Department, Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, 611731, China. muyd@uestc.edu.cn.

Abstract

Background and objective: As an important mediator of intercellular interaction and formation of extracellular bone matrix, porous scaffolds are widely used for bone regeneration. Accumulating evidences demonstrate that microRNA are involved in the regulation of scaffolds-induced bone regeneration. Recently, we revealed that miR-210-3p was highly expressed during osteogenesis induced by HAG. In present study, we further explored the molecular mechanism underlying the effect of miR-210-3p on osteogenic differentiation.

Materials and methods: In this study, miR-210-3p mimics and inhibitors were synthesized and transfected into MC3T3-E1 cells to explore their effects on osteogenic differentiation. The expression of osteogenic marker (Alp and Runx2) were detected by real-time quantitative PCR (qRT-PCR) and western blotting. After osteogenesis induction for 7 days, Alp staining were used to detected osteoblast differentiation of MC3T3-E1 cells. CCK8 and Transwell assays were performed to detected cell proliferation and migration. Then, top ranking list of target genes of miR-210-3p obtained from TargetScan and the expression of BDNF were detected by qRT-PCR and ELISA. The relationship between miR-210-3p and BDNF was verified by luciferase report assay. Furthermore, the effect of BDNF on osteoblast differentiation was verified by transfecting siRNA or adding BDNF to the culture medium.

Results: MiR-210-3p mimics markedly suppress osteogenic differentiation, cell migration and cell proliferation of MC3T3-E; nevertheless, silencing of miR-210-3p dramatically enhanced MC3T3-E1 osteogenesis, cell migration and proliferation. Furthermore, luciferase reporter assay verified that brain derived neurotrophic factor (BDNF) is a directly target of miR-210-3p. Moreover, BDNF siRNA significantly decreased the expression levels of ALP and cell migration. The addition of BDNF partially rescued the inhibition of osteogenesis by miR-210-3p.

Conclusion: miR-210-3p inhibited the osteogenic differentiation via targeting BDNF. Our Results provide a promising target for regulating osteogenic differentiation.

Keywords: BDNF; Cell migration; Osteogenic differentiation; miR-210-3p.

MeSH terms

Brain-Derived Neurotrophic Factor / genetics
Cell Differentiation / genetics
MicroRNAs* / metabolism
Osteogenesis* / genetics
RNA, Small Interfering

Substances

Brain-Derived Neurotrophic Factor
MicroRNAs
RNA, Small Interfering

Abstract

MeSH terms

Substances

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