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Analysis of HDAC1-mediated regulation of Runx2-induced osteopontin gene expression in C3h10t1/2 cells

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Abstract

Histone deacetylases (HDACs) deacetylate lysine residues of histone and non-histone proteins and thereby regulate the cell-cycle, gene expression, and several other processes. We have analyzed the effects of HDAC1 on Runx2-mediated regulation of osteopontin (OPN) promoter activation and gene expression in mesenchymal progenitor C3h10t1/2 cells and show that co-expression of HDAC1 along with Runx2 results in down-regulation of Runx2-induced OPN mRNA expression during both the proliferation and differentiation stages of C3h10t1/2 cells. Luciferase assay results revealed that HDAC1 efficiently down-regulated Runx2-stimulated OPN promoter activity in a dose-dependent manner whereas TSA relieved the HDAC1-mediated repression and up-regulated the Runx2-induced OPN promoter activity and mRNA expression. In vivo HDAC1 co-localized and physically interacted with Runx2 and associated with the OPN promoter. Thus, HDAC1 not only plays a critical role in regulation of Runx2-stimulated expression of osteogenic genes, like OPN, but also regulate the proliferation and differentiation stages of mesenchymal progenitor cells, such as C3h10t1/2.

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References

  • Byers BA, Pavlath GK, Murphy TJ, Karsenty G, Garcia AJ (2002) Cell-type-dependent up-regulation of in vitro mineralization after overexpression of the osteoblast-specific transcription factor Runx2/Cbfal. J Bone Miner Res 17(11):1931–1944

    Article  PubMed  CAS  Google Scholar 

  • Date T, Doiguchi Y, Nobuta M, Shindo H (2004) Bone morphogenetic protein-2 induces differentiation of multipotent C3H10T1/2 cells into osteoblasts, chondrocytes, and adipocytes in vivo and in vitro. J Orthop Sci 9(5):503–508

    Article  PubMed  CAS  Google Scholar 

  • Deepak V, Zhang Z, Meng L, Zeng X, Liu W (2011) Reduced activity and cytoplasmic localization of Runx2 is observed in C3h10t1/2 cells overexpressing Tbx3. Cell Biochem Funct 29(4):348–350. doi:101002/cbf1753

    Article  PubMed  CAS  Google Scholar 

  • Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89(5):747–754

    Article  PubMed  CAS  Google Scholar 

  • Franzen A, Heinegard D (1985) Isolation and characterization of two sialoproteins present only in bone calcified matrix. Biochem J 232(3):715–724

    PubMed  CAS  Google Scholar 

  • Glozak MA, Sengupta N, Zhang X, Seto E (2005) Acetylation and deacetylation of non-histone proteins. Gene 363:15–23

    Article  PubMed  CAS  Google Scholar 

  • Gravallese EM (2003) Osteopontin: a bridge between bone and the immune system. J Clin Invest 112(2):147–149

    PubMed  CAS  Google Scholar 

  • Gruber HE, Ivey JL, Thompson ER, Chesnut CH 3rd, Baylink DJ (1986) Osteoblast and osteoclast cell number and cell activity in postmenopausal osteoporosis. Miner Electrolyte Metab 12(4):246–254

    PubMed  CAS  Google Scholar 

  • Harada H, Tagashira S, Fujiwara M, Ogawa S, Katsumata T, Yamaguchi A, Komori T, Nakatsuka M (1999) Cbfa1 isoforms exert functional differences in osteoblast differentiation. J Biol Chem 274(11):6972–6978

    Article  PubMed  CAS  Google Scholar 

  • Jensen ED, Nair AK, Westendorf JJ (2007) Histone deacetylase co-repressor complex control of Runx2 and bone formation. Crit Rev Eukaryot Gene Expr 17(3):187–196

    PubMed  CAS  Google Scholar 

  • Jensen ED, Schroeder TM, Bailey J, Gopalakrishnan R, Westendorf JJ (2008) Histone deacetylase 7 associates with Runx2 and represses its activity during osteoblast maturation in a deacetylation-independent manner. J Bone Miner Res 23(3):361–372

    Article  PubMed  CAS  Google Scholar 

  • Kang JS, Alliston T, Delston R, Derynck R (2005) Repression of Runx2 function by TGF-beta through recruitment of class II histone deacetylases by Smad3. EMBO J 24(14):2543–2555

    Article  PubMed  CAS  Google Scholar 

  • Karsenty G (2008) Transcriptional control of skeletogenesis. Annu Rev Genomics Hum Genet 9:183–196

    Article  PubMed  CAS  Google Scholar 

  • Kennedy OD, Brennan O, Rackard SM, Staines A, O’Brien FJ, Taylor D, Lee TC (2009) Effects of ovariectomy on bone turnover, porosity, and biomechanical properties in ovine compact bone 12 months postsurgery. J Orthop Res 27(3):303–309

    Article  PubMed  Google Scholar 

  • Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T (1997) Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 89(5):755–764

    Article  PubMed  CAS  Google Scholar 

  • Lee HW, Suh JH, Kim AY, Lee YS, Park SY, Kim JB (2006) Histone deacetylase 1-mediated histone modification regulates osteoblast differentiation. Mol Endocrinol 20(10):2432–2443

    Article  PubMed  CAS  Google Scholar 

  • Liu W, Toyosawa S, Furuichi T, Kanatani N, Yoshida C, Liu Y, Himeno M, Narai S, Yamaguchi A, Komori T (2001) Overexpression of Cbfa1 in osteoblasts inhibits osteoblast maturation and causes osteopenia with multiple fractures. J Cell Biol 155(1):157–166

    Article  PubMed  CAS  Google Scholar 

  • Reinholt FP, Hultenby K, Oldberg A, Heinegard D (1990) Osteopontin—a possible anchor of osteoclasts to bone. Proc Natl Acad Sci U S A 87(12):4473–4475

    Article  PubMed  CAS  Google Scholar 

  • Schroeder TM, Kahler RA, Li X, Westendorf JJ (2004) Histone deacetylase 3 interacts with runx2 to repress the osteocalcin promoter and regulate osteoblast differentiation. J Biol Chem 279(40):41998–42007

    Article  PubMed  CAS  Google Scholar 

  • Westendorf JJ (2007) Histone deacetylases in control of skeletogenesis. J Cell Biochem 102(2):332–340

    Article  PubMed  CAS  Google Scholar 

  • Westendorf JJ, Zaidi SK, Cascino JE, Kahler R, van Wijnen AJ, Lian JB, Yoshida M, Stein GS, Li X (2002) Runx2 (Cbfa1, AML-3) interacts with histone deacetylase 6 and represses the p21(CIP1/WAF1) promoter. Mol Cell Biol 22(22):7982–7992

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

W. Liu thanks Dr. T. Fujita for his encouragement. This study was supported by the Training Fund of NENU’S Scientific Innovation Project (NENU-STC08015) and partially supported by The Jilin Province Science and Technology Development Project (20040114).

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Correspondence to Wenguang Liu.

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Zhongli Zhang and Vishwa Deepak are contributed equally to this work.

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Zhang, Z., Deepak, V., Meng, L. et al. Analysis of HDAC1-mediated regulation of Runx2-induced osteopontin gene expression in C3h10t1/2 cells. Biotechnol Lett 34, 197–203 (2012). https://doi.org/10.1007/s10529-011-0756-8

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  • DOI: https://doi.org/10.1007/s10529-011-0756-8

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