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Chondrocyte p21WAF1/CIP1 Expression Is Increased by Dexamethasone but Does Not Contribute to Dexamethasone-Induced Growth Retardation In Vivo

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Abstract

It has been shown that cell cycle genes play an important role in the coordination of chondrocyte proliferation and differentiation. The inhibitory effects of glucocorticoids (GCs) on chondrocyte proliferation are consistent with GCs disrupting cell cycle progression and promoting cell cycle exit. Cyclin-dependent kinase inhibitors (CDKIs) force cells to exit the cell cycle and differentiate, and studies have shown that expression of the CDKI p21CIP1/WAF1 is increased in terminally differentiated cells. In this study, p21 mRNA and protein expression was increased during chondrocyte differentiation and after exposure to dexamethasone (Dex, 10−6 M) in murine chondrogenic ATDC5 cells. In 4-week-old mice lacking a functional p21 gene, Dex caused a reduction in body weight compared to saline control null mice, but this was consistent with the reduction in body weight observed in Dex-treated wild-type littermates. In addition, p21 ablation had no effect on the reduction in width of the growth plate or reduced mineral apposition rate in Dex-treated mice. However, an alteration in growth rate and epiphyseal structure is evident when comparing p21−/− and wild-type mice. These findings suggest that p21 does not directly contribute to GC-induced growth retardation in vivo but is involved in the maintenance of the growth plate.

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References

  1. Sanchez CP, He YZ (2002) Alterations in the growth plate cartilage of rats with renal failure receiving corticosteroid therapy. Bone 30:692–698

    Article  PubMed  CAS  Google Scholar 

  2. Siebler T, Robson H, Shalet SM, Williams GR (2002) Dexamethasone inhibits and thyroid hormone promotes differentiation of mouse chondrogenic ATDC5 cells. Bone 31:457–464

    Article  PubMed  CAS  Google Scholar 

  3. Mushtaq T, Farquharson C, Seawright E, Ahmed SF (2002) Glucocorticoid effects on chondrogenesis, differentiation and apoptosis in the murine ATDC5 chondrocyte cell line. J Endocrinol 175:705–713

    Article  PubMed  CAS  Google Scholar 

  4. Johnson DG, Walker CL (1999) Cyclins and cell cycle checkpoints. Annu Rev Pharmacol Toxicol 39:295–312

    Article  PubMed  CAS  Google Scholar 

  5. Stewart MC, Kadlcek RM, Robbins PD, MacLeod JN, Ballock RT (2004) Expression and activity of the CDK inhibitor p57Kip2 in chondrocytes undergoing hypertrophic differentiation. J Bone Miner Res 19:123–132

    Article  PubMed  CAS  Google Scholar 

  6. Zhang P, Liegeois NJ, Wong C, Finegold M, Hou H, Thompson JC, Silverman A, Harper JW, DePinho RA, Elledge SJ (1997) Altered cell differentiation and proliferation in mice lacking p57KIP2 indicates a role in Beckwith–Wiedemann syndrome. Nature 387:151–158

    Article  PubMed  CAS  Google Scholar 

  7. Fero ML, Rivkin M, Tasch M, Porter P, Carow CE, Firpo E, Polyak K, Tsai LH, Broudy V, Perlmutter RM, Kaushansky K, Roberts JM (1996) A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(Kip1)-deficient mice. Cell 85:733–744

    Article  PubMed  CAS  Google Scholar 

  8. Kiyokawa H, Koff A (1998) Roles of cyclin-dependent kinase inhibitors: lessons from knockout mice. Curr Top Microbiol Immunol 227:105–120

    PubMed  CAS  Google Scholar 

  9. Deng C, Zhang P, Harper JW, Elledge SJ, Leder P (1995) Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82:675–684

    Article  PubMed  CAS  Google Scholar 

  10. Missero C, Di CF, Kiyokawa H, Koff A, Dotto GP (1996) The absence of p21Cip1/WAF1 alters keratinocyte growth and differentiation and promotes ras-tumor progression. Genes Dev 10:3065–3075

    Article  PubMed  CAS  Google Scholar 

  11. Ballock RT, Zhou X, Mink LM, Chen DH, Mita BC, Stewart MC (2000) Expression of cyclin-dependent kinase inhibitors in epiphyseal chondrocytes induced to terminally differentiate with thyroid hormone. Endocrinology 141:4552–4557

    Article  PubMed  CAS  Google Scholar 

  12. Stewart MC, Farnum CE, MacLeod JN (1997) Expression of p21CIP1/WAF1 in chondrocytes. Calcif Tissue Int 61:199–204

    Article  PubMed  CAS  Google Scholar 

  13. Zenmyo M, Komiya S, Hamada T, Hiraoka K, Suzuki R, Inoue A (2000) p21 and parathyroid hormone-related peptide in the growth plate. Calcif Tissue Int 67:378–381

    Article  PubMed  CAS  Google Scholar 

  14. Aszodi A, Hunziker EB, Brakebusch C, Fässler R (2003) β1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis. Genes Dev 17:2465–2479

    Article  PubMed  CAS  Google Scholar 

  15. Nakajima M, Negishi Y, Tanaka H, Kawashima K (2004) p21(Cip-1/SDI-1/WAF-1) expression via the mitogen-activated protein kinase signalling pathway in insulin-induced chondrogenic differentiation of ATDC5 cells. Biochem Biophys Res Commun 320:1069–1075

    Article  PubMed  CAS  Google Scholar 

  16. Macleod KF, Sherry N, Hannon G, Beach D, Tokino T, Kinzler K, Vogelstein B, Jacks T (1995) p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage. Genes Dev 9:935–944

    Article  PubMed  CAS  Google Scholar 

  17. Harper JW, Elledge SJ (1996) Cdk inhibitors in development and cancer. Curr Opin Genet Dev 6:56–64

    Article  PubMed  CAS  Google Scholar 

  18. Mushtaq T, Ahmed SF (2002) The impact of corticosteroids on growth and bone health. Arch Dis Child 87:93–96

    Article  PubMed  CAS  Google Scholar 

  19. Ahmed SF, Tucker P, Mushtaq T, Wallace AM, Williams DM, Hughes IA (2002) Short-term effects on linear growth and bone turnover in children randomized to receive prednisolone or dexamethasone. Clin Endocrinol 57:185–191

    Article  CAS  Google Scholar 

  20. Negishi Y, Ui N, Nakajima M, Kawashima K, Maruyama K, Takizawa T, Endo H (2001) p21Cip-1/SDI-1/WAF-1 gene is involved in chondrogenic differentiation of ATDC5 cells in vitro. J Biol Chem 276:33249–33256

    Article  PubMed  CAS  Google Scholar 

  21. Leclerc N, Luppen CA, Ho VV, Nagpal S, Hacia JG, Smith E, Frenkel B (2004) Gene expression profiling of glucocorticoid-inhibited osteoblasts. J Mol Endocrinol 33:175–193

    Article  PubMed  CAS  Google Scholar 

  22. Cram EJ, Ramos RA, Wang EC, Cha HH, Nishio Y, Firestone GL (1998) Role of the CCAAT/enhancer binding protein-alpha transcription factor in the glucocorticoid stimulation of p21waf1/cip1 gene promoter activity in growth-arrested rat hepatoma cells. J Biol Chem 273:2008–2014

    Article  PubMed  CAS  Google Scholar 

  23. Yang JQ, Rüdiger JJ, Hughes JM, Goulet S, Gencay-Cornelson MM, Borger P, Tamm M, Roth M (2008) Cell density and serum exposure modify the function of the glucocorticoid receptor C/EBP complex. Am J Respir Cell Mol Biol 38:414–422

    Article  PubMed  CAS  Google Scholar 

  24. James CG, Ulici V, Tuckermann J, Underhill TM, Beier F (2007) Expression profiling of dexamethasone-treated primary chondrocytes identifies targets of glucocorticoid signalling in endochondral bone development. BMC Genomics 8:205

    Article  PubMed  Google Scholar 

  25. Owen HC, Miner JN, Ahmed SF, Farquharson C (2007) The growth plate sparing effects of the selective glucocorticoid receptor modulator, AL-438. Mol Cell Endocrinol 264:164–170

    Article  PubMed  CAS  Google Scholar 

  26. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  27. Chrysis D, Ritzen EM, Savendahl L (2003) Growth retardation induced by dexamethasone is associated with increased apoptosis of the growth plate chondrocytes. J Endocrinol 176:331–337

    Article  PubMed  CAS  Google Scholar 

  28. Jacquet R, Hillyer J, Landis WJ (2005) Analysis of connective tissues by laser capture microdissection and reverse transcriptase–polymerase chain reaction. Anal Biochem 33:22–34

    Article  Google Scholar 

  29. Luo Y, Hurwitz J, Massague J (1995) Cell-cycle inhibition by independent CDK and PCNA binding domains in p21Cip1. Nature 375:159–161

    Article  PubMed  CAS  Google Scholar 

  30. Sherr CJ (1995) Mammalian G1 cyclins and cell cycle progression. Proc Assoc Am Physicians 107:181–186

    PubMed  CAS  Google Scholar 

  31. Owen HC, Roberts SJ, Ahmed SF, Farquharson C (2008) Dexamethasone-induced expression of the glucocorticoids response gene lipocalin 2 in chondrocytes. Am J Physiol Endocrinol Metab 294:E1023–E1034

    Article  PubMed  CAS  Google Scholar 

  32. Ramalingam A, Hirai A, Thompson EA (1997) Glucocorticoid inhibition of fibroblast proliferation and regulation of the cyclin kinase inhibitor p21Cip1. Mol Endocrinol 11:577–586

    Article  PubMed  CAS  Google Scholar 

  33. Corroyer S, Nabeyrat E, Clement A (1997) Involvement of the cell cycle inhibitor CIP1/WAF1 in lung alveolar epithelial cell growth arrest induced by glucocorticoids. Endocrinology 138:3677–3685

    Article  PubMed  CAS  Google Scholar 

  34. Van Buul-Offers SC, Smink JJ, Gresnigt R, Hamers N, Koedam J, Karperien M (2005) Thyroid hormone, but not parathyroid hormone, partially restores glucocorticoid-induced growth retardation. Pediatr Nephrol 20:335–341

    Article  PubMed  Google Scholar 

  35. Emons JA, Marino R, Nilsson O, Barnes KM, Even-Zohar N, Andrade AC, Chatterjee NA, Wit JM, Karperien M, Baron J (2006) The role of p27Kip1 in the regulation of growth plate chondrocyte proliferation in mice. Pediatr Res 60:288–293

    Article  PubMed  CAS  Google Scholar 

  36. Fritz PC, Ward WE, Atkinson SA, Tenenbaum HC (1998) Tamoxifen attenuates the effects of exogenous glucocorticoid on bone formation and growth in piglets. Endocrinology 139:3399–3403

    Article  PubMed  CAS  Google Scholar 

  37. Macrae VE, Ahmed SF, Mushtaq T, Farquharson C (2007) IGF-I signaling in bone growth: inhibitory actions of dexamethasone and IL-1beta. Growth Horm IGF Res 17:435–439

    Article  PubMed  CAS  Google Scholar 

  38. Altman A, Hochberg Z, Silbermann M (1992) Interactions between growth hormone and dexamethasone in skeletal growth and bone structure of the young mouse. Calcif Tissue Int 51:298–304

    Article  PubMed  CAS  Google Scholar 

  39. Kim HJ, Zhao H, Kitaura H, Bhattacharyya S, Brewer JA, Muglia LJ, Ross FP, Teitelbaum SL (2006) Glucocorticoids suppress bone formation via the osteoclast. J Clin Invest 116:2152–2160

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Drs. Bruce Whitelaw and Douglas Vasey (Roslin Institute) for providing p21−/− mice and the Small Animal Unit at the Roslin Institute for assistance with in vivo experiments. We also thank Dr. Scott Roberts (Katholieke Universiteit Leuven) for assistance with manuscript preparation and Ms. Elaine Seawright and Mr. Matt Prideaux (Roslin Institute) for technical assistance with tissue processing. This projected was funded by the Biotechnology and Biological Sciences Research Council (BBSRC) UK through a studentship award (to H. C. O.) and Institute Strategic Programme Grant funding from the BBSRC.

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Authors and Affiliations

  1. Bone Biology Group, Division of Developmental Biology, The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Roslin, Edinburgh, Midlothian, EH25 9PS, UK

    H. C. Owen & C. Farquharson

  2. Bone and Endocrine Research Group, Royal Hospital for Sick Children, Glasgow, UK

    H. C. Owen & S. F. Ahmed

  3. Laboratory for Intensive Care Medicine, Katholieke Universiteit Leuven, Leuven, 3000, Belgium

    H. C. Owen

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  1. H. C. Owen
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  2. S. F. Ahmed
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  3. C. Farquharson
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Correspondence to C. Farquharson.

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Owen, H.C., Ahmed, S.F. & Farquharson, C. Chondrocyte p21WAF1/CIP1 Expression Is Increased by Dexamethasone but Does Not Contribute to Dexamethasone-Induced Growth Retardation In Vivo. Calcif Tissue Int 85, 326–334 (2009). https://doi.org/10.1007/s00223-009-9276-0

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  • DOI: https://doi.org/10.1007/s00223-009-9276-0

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