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Journal of Bone and Mineral Metabolism

, Volume 36, Issue 6, pp 648–660 | Cite as

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

  • Lin Deng
  • Guoli Hu
  • Lei Jin
  • Chenglong Wang
  • Hongwen Niu
Original Article

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.

Keywords

MiR-23b Runx2 TNF-α BMSCs Osteogenic differentiation 

Notes

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.

Author 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.

Compliance with ethical standards

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.

References

  1. 1.
    Gutschner T, Diederichs S (2012) The hallmarks of cancer: a long non-coding RNA point of view. RNA Biol 9:703–719CrossRefGoogle Scholar
  2. 2.
    Rosenberg N, Rosenberg O, Soudry M (2012) Osteoblasts in bone physiology-mini review. Rambam Maimonides Med J 3:e0013CrossRefGoogle Scholar
  3. 3.
    Li P, Yang YM, Sanchez S, Cui DC, Dang RJ, Wang XY, Lin QX, Wang Y, Wang C, Chen DF, Chen SY, Jiang XX, Wen N (2016) Deubiquitinase MYSM1 Is essential for normal bone formation and mesenchymal stem cell differentiation. Sci Rep 6:22211CrossRefGoogle Scholar
  4. 4.
    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147CrossRefGoogle Scholar
  5. 5.
    Zhang W, Yang N, Shi XM (2008) Regulation of mesenchymal stem cell osteogenic differentiation by glucocorticoid-induced leucine zipper (GILZ). J Biol Chem 283:4723–4729CrossRefGoogle Scholar
  6. 6.
    Duplomb L, Dagouassat M, Jourdon P, Heymann D (2007) Concise review: embryonic stem cells: a new tool to study osteoblast and osteoclast differentiation. Stem Cells 25:544–552CrossRefGoogle Scholar
  7. 7.
    Nanes MS (2003) Tumor necrosis factor-alpha: molecular and cellular mechanisms in skeletal pathology. Gene 321:1–15CrossRefGoogle Scholar
  8. 8.
    Gilbert L, He X, Farmer P, Boden S, Kozlowski M, Rubin J, Nanes MS (2000) Inhibition of osteoblast differentiation by tumor necrosis factor-alpha. Endocrinology 141:3956–3964CrossRefGoogle Scholar
  9. 9.
    Beklen A, Ainola M, Hukkanen M, Gurgan C, Sorsa T, Konttinen YT (2007) MMPs, IL-1, and TNF are regulated by IL-17 in periodontitis. J Dent Res 86:347–351CrossRefGoogle Scholar
  10. 10.
    Choy EH, Panayi GS (2001) Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 344:907–916CrossRefGoogle Scholar
  11. 11.
    Ye X, Huang H, Zhao N, Zhang J, Yang P (2016) Inhibition of Runx2 signaling by TNF-alpha in ST2 murine bone marrow stromal cells undergoing osteogenic differentiation. In vitro Cell Dev Biol Anim 52:1026–1033CrossRefGoogle Scholar
  12. 12.
    Malysheva K, de Rooij K, Lowik CW, Baeten DL, Rose-John S, Stoika R, Korchynskyi O (2016) Interleukin 6/Wnt interactions in rheumatoid arthritis: interleukin 6 inhibits Wnt signaling in synovial fibroblasts and osteoblasts. Croat Med J 57:89–98CrossRefGoogle Scholar
  13. 13.
    Thomas M, Lieberman J, Lal A (2010) Desperately seeking microRNA targets. Nat Struct Mol Biol 17:1169–1174CrossRefGoogle Scholar
  14. 14.
    Esteller M (2011) Non-coding RNAs in human disease. Nat Rev Genet 12:861–874CrossRefGoogle Scholar
  15. 15.
    Itoh T, Takeda S, Akao Y (2010) MicroRNA-208 modulates BMP-2-stimulated mouse preosteoblast differentiation by directly targeting V-ets erythroblastosis virus E26 oncogene homolog 1. J Biol Chem 285:27745–27752CrossRefGoogle Scholar
  16. 16.
    Schaap-Oziemlak AM, Raymakers RA, Bergevoet SM, Gilissen C, Jansen BJ, Adema GJ, Kögler G, le Sage C, Agami R, van der Reijden BA, Jansen JH (2010) MicroRNA hsa-miR-135b regulates mineralization in osteogenic differentiation of human unrestricted somatic stem cells. Stem Cells Dev 19:877–885CrossRefGoogle Scholar
  17. 17.
    Huang J, Zhao L, Xing L, Chen D (2010) MicroRNA-204 regulates Runx2 protein expression and mesenchymal progenitor cell differentiation. Stem Cells 28:357–364PubMedPubMedCentralGoogle Scholar
  18. 18.
    Jeong BC, Kang IH, Hwang YC, Kim SH, Koh JT (2014) MicroRNA-194 reciprocally stimulates osteogenesis and inhibits adipogenesis via regulating COUP-TFII expression. Cell Death Dis 5:e1532CrossRefGoogle Scholar
  19. 19.
    Li H, Li T, Fan J, Li T, Fan L, Wang S, Weng X, Han Q, Zhao RC (2015) miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway. Cell Death Differ 22:1935–1945CrossRefGoogle Scholar
  20. 20.
    Hu R, Li H, Liu W, Yang L, Tan YF, Luo XH (2010) Targeting miRNAs in osteoblast differentiation and bone formation. Expert Opin Ther Targets 14:1109–1120CrossRefGoogle Scholar
  21. 21.
    Guo Q, Chen Y, Guo L, Jiang T, Lin Z (2016) miR-23a/b regulates the balance between osteoblast and adipocyte differentiation in bone marrow mesenchymal stem cells. Bone Res 4:16022CrossRefGoogle Scholar
  22. 22.
    Chen C, Tang Z, Song Q, Yang M, Shi Q, Weng Y (2016) Downregulated microRNA-23b promotes BMP9-mediated osteogenesis in C2C12 myoblast cells by targeting Runx2. Mol Med Rep 13:2492–2498CrossRefGoogle Scholar
  23. 23.
    Dalle Carbonare L, Innamorati G, Valenti MT (2012) Transcription factor Runx2 and its application to bone tissue engineering. Stem Cell Rev 8:891–897CrossRefGoogle Scholar
  24. 24.
    Komori T (2010) Regulation of bone development and extracellular matrix protein genes by RUNX2. Cell Tissue Res 339:189–195CrossRefGoogle Scholar
  25. 25.
    Phimphilai M, Zhao Z, Boules H, Roca H, Franceschi RT (2006) BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res 21:637–646CrossRefGoogle Scholar
  26. 26.
    Xiao G, Jiang D, Ge C, Zhao Z, Lai Y, Boules H, Phimphilai M, Yang X, Karsenty G, Franceschi RT (2005) Cooperative interactions between activating transcription factor 4 and Runx2/Cbfa1 stimulate osteoblast-specific osteocalcin gene expression. J Biol Chem 280:30689–30696CrossRefGoogle Scholar
  27. 27.
    Lee WY, Li N, Lin S, Wang B, Lan HY, Li G (2016) miRNA-29b improves bone healing in mouse fracture model. Mol Cell Endocrinol 430:97–107CrossRefGoogle Scholar
  28. 28.
    Peng S, Gao D, Gao C, Wei P, Niu M, Shuai C (2016) MicroRNAs regulate signaling pathways in osteogenic differentiation of mesenchymal stem cells (review). Mol Med Rep 14:623–629CrossRefGoogle Scholar
  29. 29.
    Huang C, Geng J, Jiang S (2017) MicroRNAs in regulation of osteogenic differentiation of mesenchymal stem cells. Cell Tissue Res 368:229–238CrossRefGoogle Scholar
  30. 30.
    Yoshizuka M, Nakasa T, Kawanishi Y, Hachisuka S, Furuta T, Miyaki S, Adachi N, Ochi M (2016) Inhibition of microRNA-222 expression accelerates bone healing with enhancement of osteogenesis, chondrogenesis, and angiogenesis in a rat refractory fracture model. J Orthop Sci 21:852–858CrossRefGoogle Scholar

Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan KK, part of Springer Nature 2017

Authors and Affiliations

  • Lin Deng
    • 1
  • Guoli Hu
    • 2
  • Lei Jin
    • 3
  • Chenglong Wang
    • 4
  • Hongwen Niu
    • 1
  1. 1.Department of TraumatologyShu Guang Hospital Affiliated to Shanghai University of Traditional Chinese MedicineShanghaiChina
  2. 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 SciencesShanghaiChina
  3. 3.Anorectal Surgery, Shanghai Min Hang Traditional Chinese Medicine HospitalShanghaiChina
  4. 4.Department of Orthopedic SurgeryXin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (SJTUSM)ShanghaiChina

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