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Calcified Tissue International

, Volume 57, Issue 6, pp 436–441 | Cite as

Progesterone receptors are expressed in human osteoblast-like cell lines and in primary human osteoblast cultures

  • P. MacNamara
  • C. O'Shaughnessy
  • P. Manduca
  • H. C. Loughrey
Article

Abstract

Expression of progesterone receptors (PR) was studied in human osteoblast-like cell lines and primary human osteoblast cultures at the molecular level. Using the sensitive reverse transcriptase polymerase chain reaction (RT-PCR) and oligonucleotide primers which flank the progesterone-binding domain of human PR, progesterone receptor (PR) mRNA was detected in three osteoblast-like cell lines-HOS-TE85, MG-63, and SAOS-2. When compared with β-actin gene expression, levels of PRmRNA transcripts varied between cell lines (PRmRNA in HOS-TE85>MG-63 ≫ SAOS-2). In addition, RT-PCR confirmed the presence of PRmRNA transcripts in primary human osteoblast cells cultured from collagenase-treated bone. Immunostaining was used to visualize PR protein in cells. All osteoblast-like cell lines showed specific staining for PR. Immunoreactivity was distributed equally in the nucleus and cytoplasm. The level of staining was significantly lower than that detected in PR-positive MCF-7 breast cancer cells though well above background levels obtained for PR-negative HeLa cells. The finding that PR is expressed at both the level of mRNA and protein in several osteoblast-like cell lines as well as in human primary osteoblast cultures indicates that bone-forming osteoblast cells are direct targets for progesterone action.

Key words

Human osteoblast Progesterone receptor Osteoporosis 

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References

  1. 1.
    Hahn TJ, Halstead LR, Teitelbaum SL, Hahn BH (1979) Altered mineral metabolism in glucocorticoid-induced osteopenoia. J Clin Invest 64:655–665Google Scholar
  2. 2.
    Riggs BL, Melton LJ (1986) Involutional osteoporosis. N Engl J Med 314:1676Google Scholar
  3. 3.
    Eriksen EF, Colvard DS, Berg NJ, Graham ML, Mann KG, Spelsberg TC, Riggs BL (1988) Evidence of estrogen receptors in normal human osteoblast-like cells. Science 241:84–86Google Scholar
  4. 4.
    Komm BS, Terpening CM, Benz DJ, Graeme KA, Gallegos A, Kore M, Greene GL, O'Malley B, Haussler MR (1988) Estrogen binding, receptor mRNA and biological response in osteoblast-like osteosarcoma cells. Science 241:81–84Google Scholar
  5. 5.
    Prior JC (1990) Progesterone as a bone-trophic hormone. Endocrine 11:2:386–398Google Scholar
  6. 6.
    Barbieri RL (1990) Gonadotropin-releasing hormone antagonists and estrogen-progestrogen replacement therapy. Am J Obstet Gynecol 162:593–595Google Scholar
  7. 7.
    Gambrell RD Jr (1989) Uses of progestrogens in postmenopausal woman. Int J Fertil 34:315–331Google Scholar
  8. 8.
    Grecu EO, Weinshelbaum A, Simmons R (1990) Effective therapy of glucocorticoid-induced osteoporosis with methoxyprogesterone acetate. Calcif Tissue Int 46;294–299Google Scholar
  9. 9.
    Manolagues SC, Anderson DC (1978) Detection of high affinity glucocorticoid binding in the rat bone. J Endocrinol 76:379–380Google Scholar
  10. 10.
    Chen TL, Aronow L, Feldman DL (1977) Glucocorticoid receptors and inhibition of bone cell growth in primary culture. Endocrinology 100:819–628Google Scholar
  11. 11.
    Orwell ES, Stribrska L, Ramsey EE, Keenan EJ (1991) Androgen receptors in osteoblast-like cell lines. Calcif Tissue Int 49: 183–187Google Scholar
  12. 12.
    Slootweg MC Ederveen AGH, Schot LPC, Schooner WGEJ, Kloosterboer HJ, (1992) Oestrogen and progestogen synergistically stimulate human and rat osteoblast proliferation. J Endocrinol 133:R5-R8Google Scholar
  13. 13.
    Wei LL, Leach MW, Miner RS, Demers LM (1993) Evidence for progesterone receptors in human osteoblast-like cells. Biochem Biophys Res Commun 195:525–532Google Scholar
  14. 14.
    Robey P, Termine JD (1985) Human bone cells in vitro. Calcif Tissue Int 37:453–460Google Scholar
  15. 15.
    Manduca P, Sanguineti C, Pistone M, Boccignone E, Sanguineti F, Santolini F, Federici A (1993) Differential expression of alkaline phosphatase in clones of human osteoblast-like cells. J Bone Miner Res 8:291–300Google Scholar
  16. 16.
    Chomczynski P, Sacchi N (1987) Single step method of RNA isolation by acid guanidium thiocyanate-phemol-chloroform extraction. Anal Biochem 162:156–159Google Scholar
  17. 17.
    Hagihara K, Hirata S, Osada T, Hirai M, Kato J (1992) Expression of progesterone receptor in the hepnatal rat brain cortex: detection of its mRNA using reverse transcription polymerase chain reaction. J Steroid Biochem Molec Biol 41:637–640Google Scholar
  18. 18.
    Misrahi M, Atger M, et al. (1987) Complete amino acid sequence of the human progesterone receptor deduced form cloned cDNA. Biochem Biophys res Commun 143:740–748Google Scholar
  19. 19.
    Horikoshi T, Danenberg K, Stradbauer THW, et al. (1992) Quantification of thymidylate synthase, dihydrofolate reductase and DT-diaphoase expression in human tumours using the polymeraes chain reaction. Cancer Res 52:108–116Google Scholar
  20. 20.
    Rifas L, Fausto A, Scott MJ, Avioli LV, Welgus HG (1994) Expression of metalloproteinases and tissue inhibitors of metalloproteinases in human osteoblast-like cells: differentiation is associated with repression of metalloproteinase biosynthesis. Endocrinology 134(1):213–220Google Scholar
  21. 21.
    Tremollieres FA, Strong D, Baylink DJ, Mohan S (1992) Progesterone and promegestone stimulate human bone cell proliferation and insulin-like growth factor-2 production. Acta Endocrinol 126:329–337Google Scholar
  22. 22.
    Verharr HJJ, Damen CA, Duursma, SA, Scheven BAA (1994) A comparison of the action of progestins and estrogens on the growth and the differentation of normal adult human osteoblast-like cells in vitro. Bone 15:307–311Google Scholar
  23. 23.
    Pensier JM, Radosevich JA, Higbee R, Langman CB (1990) Osteoclast isolated from membranous bone in children exhibit nuclear estrogen and progesterone, receptors. J. Bone Miner Res 5:797–802Google Scholar
  24. 24.
    Kastner P, Krust A, Turcotte B, Stroop U, Tora L, Gromemeyer H, Chambon P (1990) Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. EMBO J 9:1603–1614Google Scholar
  25. 25.
    Greene GL, Harris K, Bova R, Kinders R, Moore B, and Nolan C (1988) Purification of T47D human progesterone receptor and immunocharacterization with monoclonal antibodies. Mol Endbocrinol 2:714–726Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1995

Authors and Affiliations

  • P. MacNamara
    • 1
  • C. O'Shaughnessy
    • 1
  • P. Manduca
    • 2
  • H. C. Loughrey
    • 1
  1. 1.Department of BiochemistryUniversity College GalwayGalwayIreland
  2. 2.Istituto di Fisiologia GeneticaFacolta di Scienze, M.F.N.GenoaItaly

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