Download PDF
Horm Metab Res 2009; 41(12): 861-865
DOI: 10.1055/s-0029-1233460
Original Basic

© Georg Thieme Verlag KG Stuttgart · New York

Parathyroid Hormone Upregulates BMP-2 mRNA Expression Through Mevalonate Kinase and Rho Kinase Inhibition in Osteoblastic MC3T3-E1 Cells

H. Takase1 , S. Yano1 , T. Yamaguchi1 , I. Kanazawa1 , K. Hayashi1 , M. Yamamoto1 , M. Yamauchi1 , T. Sugimoto1
  • 1Shimane University Faculty of Medicine, Department of Internal Medicine 1, Enya-cho, Izumo, Japan
Further Information

Publication History

received 01.05.2009

accepted 17.06.2009

Publication Date:
11 August 2009 (online)

Also available at
    Permissions and Reprints

Abstract

It is well known that parathyroid hormone (PTH) possesses an anabolic effect on bone. However, the mechanisms are not fully elucidated. So far, it is unclear whether or not PTH could stimulate the expression of bone morphogenetic protein-2 (BMP-2), a strong mediator for bone formation. Growing evidence suggests that BMP-2 expression is regulated by the mevalonate pathway and Rho-associated protein kinase (ROK) activity. This study was performed to examine if PTH affects BMP-2 expression and to clarify its involvement of the mevalonate pathway. Osteoblastic MC3T3-E1 cells were treated with human PTH-(1–34) to determine BMP-2 mRNA expression levels by real-time PCR and to measure the ROK activity by the kinase assay. Incubation with 10−9–10−8 M of hPTH-(1–34) for 6 h induced significant upregulation of BMP-2 mRNA levels in MC3T3-E1 cells. Short-term treatment of hPTH-(1–34) suppressed Rho kinase activity and mevalonate kinase mRNA levels. PTH-induced BMP-2 mRNA upregulation was selectively reversed by geranylgeranyl pyrophosphate (GGPP) pretreatment, but not by mevalonate pretreatment. These findings suggest that BMP-2 mRNA expression was upregulated by PTH in MC3T3-E1 cells mediated by mevalonate pathway suppression followed by ROK inhibition. We have now demonstrated for the first time that PTH stimulated BMP-2 mRNA expression via the mevalonate pathway and ROK in osteoblastic MC3T3-E1 cells.

Key words

osteoblast - parathyroid hormone - BMP-2 - Rho kinase

References

  • 1 Arlot M, Meunier PJ, Boivin G, Haddock L, Tamayo J, Correa-Rotter R, Jasqui S, Donley DW, Dalsky GP, Martin JS, Eriksen EF. Differential effects of teriparatide and alendronate on bone remodeling in postmenopausal women assessed by histomorphometric parameters.  J Bone Miner Res. 2005;  20 1244-1253
    Google Scholar
  • 2 Chen P, Satterwhite JH, Licata AA, Lewiecki EM, Sipos AA, Misurski DM, Wagman RB. Early changes in biochemical markers of bone formation predict BMD response to teriparatide in postmenopausal women with osteoporosis.  J Bone Miner Res. 2005;  20 962-970
    Google Scholar
  • 3 Dobnig H, Sipos A, Jiang Y, Fahrleitner-Pammer A, Ste-Marie LG, Gallagher JC, Pavo I, Wang J, Eriksen EF. Early changes in biochemical markers of bone formation correlate with improvements in bone structure during teriparatide therapy.  J Clin Endocrinol Metab. 2005;  90 3970-3977
    Google Scholar
  • 4 Jilka RL, Weinstein RS, Bellido T, Roberson P, Parfitt AM, Manolagas SC. Increased bone formation by prevention of osteoblast apoptosis with parathyroid hormone.  J Clin Invest. 1999;  104 439-446
    Google Scholar
  • 5 Nishida S, Yamaguchi A, Tanizawa T, Endo N, Mashiba T, Uchiyama Y, Suda T, Yoshiki S, Takahashi HE. Increased bone formation by intermittent parathyroid hormone administration is due to the stimulation of proliferation and differentiation of osteoprogenitor cells in bone marrow.  Bone. 1994;  15 717-723
    Google Scholar
  • 6 Bellido T, Ali AA, Plotkin LI, Fu Q, Gubrij I, Roberson PK, Weinstein RS, O’Brien CA, Manolagas SC, Jilka RL. Proteasomal degradation of Runx2 shortens parathyroid hormone-induced anti-apoptotic signaling in osteoblasts. A putative explanation for why intermittent administration is needed for bone anabolism.  J Biol Chem. 2003;  278 50259-50272
    Google Scholar
  • 7 Miyakoshi N, Kasukawa Y, Linkhart TA, Baylink DJ, Mohan S. Evidence that anabolic effects of PTH on bone require IGF-I in growing mice.  Endocrinology. 2001;  142 4349-4356
    Google Scholar
  • 8 Tobimatsu T, Kaji H, Sowa H, Naito J, Canaff L, Hendy GN, Sugimoto T, Chihara K. Parathyroid hormone increases beta-catenin levels through Smad3 in mouse osteoblastic cells.  Endocrinology. 2006;  147 2583-2590
    Google Scholar
  • 9 Kulkarni NH, Halladay DL, Miles RR, Gilbert LM, Frolik CA, Galvin RJ, Martin TJ, Gillespie MT, Onyia JE. Effects of parathyroid hormone on Wnt signaling pathway in bone.  J Cell Biochem. 2005;  95 1178-1190
    Google Scholar
  • 10 McCarthy TL, Centrella M, Canalis E. Parathyroid hormone enhances the transcript and polypeptide levels of insulin-like growth factor I in osteoblast-enriched cultures from fetal rat bone.  Endocrinology. 1989;  124 1247-1253
    Google Scholar
  • 11 Bikle DD, Sakata T, Leary C, Elalieh H, Ginzinger D, Rosen CJ, Beamer W, Majumdar S, Halloran BP. Insulin-like growth factor I is required for the anabolic actions of parathyroid hormone on mouse bone.  J Bone Miner Res. 2002;  17 1570-1578
    Google Scholar
  • 12 Reddi AH. Bone morphogenetic proteins: from basic science to clinical applications.  J Bone Joint Surg Am. 2001;  83-A ((Suppl 1)) S1-S6
    Google Scholar
  • 13 Einhorn TA, Majeska RJ, Mohaideen A, Kagel EM, Bouxsein ML, Turek TJ, Wozney JM. A single percutaneous injection of recombinant human bone morphogenetic protein-2 accelerates fracture repair.  J Bone Joint Surg Am. 2003;  85-A 1425-1435
    Google Scholar
  • 14 Issack PS, DiCesare PE. Recent advances toward the clinical application of bone morphogenetic proteins in bone and cartilage repair.  Am J Orthop. 2003;  32 429-436
    Google Scholar
  • 15 Wozney JM, Rosen V. Bone morphogenetic protein and bone morphogenetic protein gene family in bone formation and repair.  Clin Orthop Relat Res. 1998;  346 26-37
    Google Scholar
  • 16 Khosla S, Westendorf JJ, Oursler MJ. Building bone to reverse osteoporosis and repair fractures.  J Clin Invest. 2008;  118 421-428
    Google Scholar
  • 17 Hayashi K, Yamaguchi T, Yano S, Kanazawa I, Yamauchi M, Yamamoto M, Sugimoto T. BMP/Wnt antagonists are upregulated by dexamethasone in osteoblasts and reversed by alendronate and PTH: potential therapeutic targets for glucocorticoid-induced osteoporosis.  Biochem Biophys Res Commun. 2009;  379 261-266
    Google Scholar
  • 18 Laufs U, Liao JK. Post-transcriptional regulation of endothelial nitric oxide synthase mRNA stability by Rho GTPase.  J Biol Chem. 1998;  273 24266-24271
    Google Scholar
  • 19 Mundy G, Garrett R, Harris S, Chan J, Chen D, Rossini G, Boyce B, Zhao M, Gutierrez G. Stimulation of bone formation in vitro and in rodents by statins.  Science. 1999;  286 1946-1949
    Google Scholar
  • 20 Laufs U, Fata VL, Liao JK. Inhibition of 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase blocks hypoxia-mediated down-regulation of endothelial nitric oxide synthase.  J Biol Chem. 1997;  272 31725-31729
    Google Scholar
  • 21 Garrett IR, Gutierrez G, Mundy GR. Statins and bone formation.  Curr Pharm Des. 2001;  7 715-736
    Google Scholar
  • 22 Goldstein JL, Brown MS. Regulation of the mevalonate pathway.  Nature. 1990;  343 425-430
    Google Scholar
  • 23 Lee MH, Cho YS, Han YM. Simvastatin suppresses self-renewal of mouse embryonic stem cells by inhibiting RhoA geranylgeranylation.  Stem Cells. 2007;  25 1654-1663
    Google Scholar
  • 24 Kanazawa I, Yamaguchi T, Yano S, Yamauchi M, Sugimoto T. Activation of AMP-kinase and inhibition of Rho-kinase induce the mineralization of osteoblastic MC3T3-E1 cells through endothelial NOS and BMP-2 expression.  Am J Physiol Endocrinol Metab. 2009;  296 E139-E146
    Google Scholar
  • 25 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method.  Methods. 2001;  25 402-408
    Google Scholar
  • 26 Yano S, Mentaverri R, Kanuparthi D, Bandyopadhyay S, Rivera A, Brown EM, Chattopadhyay N. Functional expression of beta-chemokine receptors in osteoblasts: role of regulated upon activation, normal T cell expressed and secreted (RANTES) in osteoblasts and regulation of its secretion by osteoblasts and osteoclasts.  Endocrinology. 2005;  146 2324-2335
    Google Scholar
  • 27 Swarthout JT, D’Alonzo RC, Selvamurugan N, Partridge NC. Parathyroid hormone-dependent signaling pathways regulating genes in bone cells.  Gene. 2002;  282 1-17
    Google Scholar
  • 28 Bellido T, Ali AA, Gubrij I, Plotkin LI, Fu Q, O’Brien CA, Manolagas SC, Jilka RL. Chronic elevation of parathyroid hormone in mice reduces expression of sclerostin by osteocytes: a novel mechanism for hormonal control of osteoblastogenesis.  Endocrinology. 2005;  146 4577-4583
    Google Scholar
  • 29 Qin L, Qiu P, Wang L, Li X, Swarthout JT, Soteropoulos P, Tolias P, Partridge NC. Gene expression profiles and transcription factors involved in parathyroid hormone signaling in osteoblasts revealed by microarray and bioinformatics.  J Biol Chem. 2003;  278 19723-19731
    Google Scholar

Correspondence

S. Yano

Department of Internal Medicine 1

Shimane University Faculty of Medicine

89-1 Enya-cho

693-8501 Izumo

Japan

Phone: +81/853/20 21 83

Fax: +81/853/23 86 50

Email: syano@med.shimane-u.ac.jp