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Involvement of microRNA-23b in TNF-α-reduced BMSC osteogenic differentiation via targeting runx2

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Abstract

Elucidation of the molecular mechanism governing bone marrow mesenchymal stem cell (BMSC) osteogenic differentiation is of great importance for improving the treatment of osteoporosis. TNF-α is a well-known inhibitory factor during osteogenic differentiation of BMSCs. In our experiment, we consistently observed that TNF-α significantly inhibited BMSC osteogenic differentiation, which was partially rescued by BAY 11-7082 (NF-κB inhibitor). In this study, we examined the potential roles of microRNAs (miRNAs) involved in TNF-α-mediated reduction of BMSC osteogenesis. We found that microRNA-23b (miR-23b) was dramatically induced under the stimulation of TNF-α, which was abolished by BAY 11-7082. Similar to the effect of TNF-α, miR-23b agonist (agomir-23b) obviously impaired BMSC osteogenic differentiation in vitro and in vivo. However, agomir-23b had no effect on osteoclast activity. Overexpression of miR-23b significantly reduced runx2, the master transcription factor during osteogenesis, suggesting that miR-23b acts as an endogenous attenuator of runx2 in BMSCs. Mutation of the putative miR-23b binding site in runx2 mRNA blocked miR-23b-mediated repression of the runx2 3′ untranslated region (3′UTR) luciferase reporter activity, suggesting that miR-23b directly binds to runx2 3′UTR. Furthermore, infection with Ad-runx2 (adenovirus carrying the entire CDS sequence of runx2) effectively rescued the inhibition of BMSC osteogenic differentiation in miR-23b-overexpressing cells, indicating that the inhibiting effect of miR-23b on osteogenesis is mediated by suppression of runx2. Moreover, caudal vein injection of agomir-23b notably caused severe osteoporosis in mice, and forced expression of runx2 by combined injecting Ad-runx2 attenuated the bone loss induced by miR-23b. Collectively, these data indicated that miR-23b was involved in TNF-α-mediated reduction of BMSC osteogenesis by targeting runx2. These findings may provide new insights into understanding the regulatory role of miR-23b in the process of BMSC osteogenic differentiation in inflammatory conditions and a novel therapeutic target for osteoporosis.

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Acknowledgements

We thank Dr. Bin Zuo (Department of Orthopedic Surgery, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China) for providing human bone marrow blood samples from patients undergoing joint operation.

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Author notes

  1. Lin Deng and Guo-Li Hu contributed equally to this work.

Authors and Affiliations

  1. Department of Traumatology, Shu Guang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China

    Lin Deng & Hongwen Niu

  2. Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Jiao Tong University School of Medicine (SJTUSM) and Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences, Shanghai, China

    Guoli Hu

  3. Anorectal Surgery, Shanghai Min Hang Traditional Chinese Medicine Hospital, Shanghai, China

    Lei Jin

  4. Department of Orthopedic Surgery, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai, China

    Chenglong Wang

Authors
  1. Lin Deng
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  2. Guoli Hu
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  3. Lei Jin
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  4. Chenglong Wang
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  5. Hongwen Niu
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Contributions

LD and GH designed the study, performed the experiments, analysed the data and wrote the manuscript. LJ and CW performed the experiments and analysed the data. H-WN conceived the study, revised the manuscript and donated supplies and/or funding.

Corresponding author

Correspondence to Hongwen Niu.

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Funding

This work was supported by grants from the Key Discipline Construction Fund of Shanghai Education Commission (J50206).

Conflict of interest

The authors declare no competing financial interests.

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Deng, L., Hu, G., Jin, L. et al. Involvement of microRNA-23b in TNF-α-reduced BMSC osteogenic differentiation via targeting runx2. J Bone Miner Metab 36, 648–660 (2018). https://doi.org/10.1007/s00774-017-0886-8

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  • DOI: https://doi.org/10.1007/s00774-017-0886-8

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