Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1986 Jun;77(6):1897–1902. doi: 10.1172/JCI112517

Effects of DNA and prostaglandin synthesis inhibitors on the stimulation of bone resorption by epidermal growth factor in fetal rat long-bone cultures.

J A Lorenzo, J Quinton, S Sousa, L G Raisz
PMCID: PMC370549  PMID: 3086384

Abstract

We examined two inhibitors of DNA synthesis, hydroxyurea (HU) and aphidicholin (APC), and two inhibitors of prostaglandin cyclooxygenase, indomethacin and flufenamic acid, for their effects on the resorptive responses of fetal rat long-bone cultures to epidermal growth factor (EGF) and parathyroid hormone (PTH). As we have previously found, HU decreased unstimulated 45Ca release but had little effect on the resorptive response to PTH. HU also did not block resorption stimulated by EGF. Addition of the cyclooxygenase inhibitor, indomethacin, did not alter the resorptive responses of unstimulated or PTH-treated cultures in either the presence or absence of HU or the resorptive response of bones cultured with EGF alone. However, indomethacin completely blocked the resorptive response to EGF of bones that were cultured with HU. The effects of indomethacin on EGF-mediated resorption in HU-treated cultures appeared to be related to an inhibition of prostaglandin synthesis since flufenamic acid had similar effects. However, the effects of HU on the resorptive response to EGF may not have resulted solely from its inhibitory action on DNA synthesis since APC, in the absence of cyclooxygenase inhibitors, completely blocked EGF-mediated resorption without significantly affecting the response to PTH. These results demonstrate that the mechanisms regulating PTH- and EGF-mediated resorption in fetal rat long-bone cultures differ, and imply that a component of EGF-mediated resorption in these cultures is dependent on sustained DNA synthesis.

Full text

PDF
1897

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brune K., Rainsford K. D., Wagner K., Peskar B. A. Inhibition of anti-inflammatory drugs of prostaglandin production in cultured macrophages. Naunyn Schmiedebergs Arch Pharmacol. 1981 Jan;315(3):269–276. doi: 10.1007/BF00499844. [DOI] [PubMed] [Google Scholar]
  2. Brunk C. F., Jones K. C., James T. W. Assay for nanogram quantities of DNA in cellular homogenates. Anal Biochem. 1979 Jan 15;92(2):497–500. doi: 10.1016/0003-2697(79)90690-0. [DOI] [PubMed] [Google Scholar]
  3. Canalis E., Raisz L. G. Effect of epidermal growth factor on bone formation in vitro. Endocrinology. 1979 Apr;104(4):862–869. doi: 10.1210/endo-104-4-862. [DOI] [PubMed] [Google Scholar]
  4. Charnay P., Maniatis T. Transcriptional regulation of globin gene expression in the human erythroid cell line K562. Science. 1983 Jun 17;220(4603):1281–1283. doi: 10.1126/science.6574602. [DOI] [PubMed] [Google Scholar]
  5. Chen T. L., Cone C. M., Feldman D. Glucocorticoid modulation of cell proliferation in cultured osteoblast-like bone cells: differences between rat and mouse. Endocrinology. 1983 May;112(5):1739–1745. doi: 10.1210/endo-112-5-1739. [DOI] [PubMed] [Google Scholar]
  6. Cohen S., Taylor J. M. Epidermal growth factor: chemical and biological characterization. Recent Prog Horm Res. 1974;30(0):533–550. doi: 10.1016/b978-0-12-571130-2.50017-1. [DOI] [PubMed] [Google Scholar]
  7. Earp S. Epidermal growth factor receptor--yet another domain? Nature. 1984 May 17;309(5965):209–210. doi: 10.1038/309209a0. [DOI] [PubMed] [Google Scholar]
  8. Gregory H. Isolation and structure of urogastrone and its relationship to epidermal growth factor. Nature. 1975 Sep 25;257(5524):325–327. doi: 10.1038/257325a0. [DOI] [PubMed] [Google Scholar]
  9. Hong S. L., Polsky-Cynkin R., Levine L. Stimulation of prostaglandin biosynthesis by vasoactive substances in methylcholanthrene-transformed mouse BALB/3T3. J Biol Chem. 1976 Feb 10;251(3):776–780. [PubMed] [Google Scholar]
  10. Huot R. I., Foidart J. M., Nardone R. M., Stromberg K. Differential modulation of human chorionic gonadotropin secretion by epidermal growth factor in normal and malignant placental cultures. J Clin Endocrinol Metab. 1981 Nov;53(5):1059–1063. doi: 10.1210/jcem-53-5-1059. [DOI] [PubMed] [Google Scholar]
  11. Ibbotson K. J., D'Souza S. M., Ng K. W., Osborne C. K., Niall M., Martin T. J., Mundy G. R. Tumor-derived growth factor increases bone resorption in a tumor associated with humoral hypercalcemia of malignancy. Science. 1983 Sep 23;221(4617):1292–1294. doi: 10.1126/science.6577602. [DOI] [PubMed] [Google Scholar]
  12. Ibbotson K. J., D'Souza S. M., Smith D. D., Carpenter G., Mundy G. R. EGF receptor antiserum inhibits bone resorbing activity produced by a rat Leydig cell tumor associated with the humoral hypercalcemia of malignancy. Endocrinology. 1985 Jan;116(1):469–471. doi: 10.1210/endo-116-1-469. [DOI] [PubMed] [Google Scholar]
  13. Ibbotson K. J., Twardzik D. R., D'Souza S. M., Hargreaves W. R., Todaro G. J., Mundy G. R. Stimulation of bone resorption in vitro by synthetic transforming growth factor-alpha. Science. 1985 May 24;228(4702):1007–1009. doi: 10.1126/science.3859011. [DOI] [PubMed] [Google Scholar]
  14. Lee L. S., Weinstein I. B. Epidermal growth factor, like phorbol esters, induces plasminogen activator in HeLa cells. Nature. 1978 Aug 17;274(5672):696–697. doi: 10.1038/274696a0. [DOI] [PubMed] [Google Scholar]
  15. Letvin N. L., Linch D. C., Beardsley G. P., McIntyre K. W., Nathan D. G. Augmentation of fetal-hemoglobin production in anemic monkeys by hydroxyurea. N Engl J Med. 1984 Apr 5;310(14):869–873. doi: 10.1056/NEJM198404053101401. [DOI] [PubMed] [Google Scholar]
  16. Levine L., Hassid A. Epidermal growth factor stimulates prostaglandin biosynthesis by canine kidney (MDCK) cells. Biochem Biophys Res Commun. 1977 Jun 20;76(4):1181–1187. doi: 10.1016/0006-291x(77)90980-9. [DOI] [PubMed] [Google Scholar]
  17. Lorenzo J. A., Raisz L. G., Hock J. M. DNA synthesis is not necessary for osteoclastic responses to parathyroid hormone in cultured fetal rat long bones. J Clin Invest. 1983 Dec;72(6):1924–1929. doi: 10.1172/JCI111156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marquardt H., Hunkapiller M. W., Hood L. E., Twardzik D. R., De Larco J. E., Stephenson J. R., Todaro G. J. Transforming growth factors produced by retrovirus-transformed rodent fibroblasts and human melanoma cells: amino acid sequence homology with epidermal growth factor. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4684–4688. doi: 10.1073/pnas.80.15.4684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Masuda H., Yamamoto M., Inokuchi K., Endo H. Hydroxyurea inhibits degradation of internalized epidermal growth factor in HeLa cells. Biochem Biophys Res Commun. 1982 Nov 16;109(1):202–209. doi: 10.1016/0006-291x(82)91585-6. [DOI] [PubMed] [Google Scholar]
  20. Mundy G. R., Ibbotson K. J., D'Souza S. M. Tumor products and the hypercalcemia of malignancy. J Clin Invest. 1985 Aug;76(2):391–394. doi: 10.1172/JCI111984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ng K. W., Partridge N. C., Niall M., Martin T. J. Stimulation of DNA synthesis by epidermal growth factor in osteoblast-like cells. Calcif Tissue Int. 1983 Jul;35(4-5):624–628. doi: 10.1007/BF02405105. [DOI] [PubMed] [Google Scholar]
  22. Platt O. S., Orkin S. H., Dover G., Beardsley G. P., Miller B., Nathan D. G. Hydroxyurea enhances fetal hemoglobin production in sickle cell anemia. J Clin Invest. 1984 Aug;74(2):652–656. doi: 10.1172/JCI111464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. RAISZ L. G. BONE RESORPTION IN TISSUE CULTURE. FACTORS INFLUENCING THE RESPONSE TO PARATHYROID HORMONE. J Clin Invest. 1965 Jan;44:103–116. doi: 10.1172/JCI105117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Raisz L. G., Lorenzo J., Gworek S., Kream B., Rosenblatt M. Comparison of the effects of a potent synthetic analog of bovine parathyroid hormone with native bPTH-(1-84) and synthetic bPTH-(1-34) on bone resorption and collagen synthesis. Calcif Tissue Int. 1979;29(3):215–218. doi: 10.1007/BF02408083. [DOI] [PubMed] [Google Scholar]
  25. Raisz L. G., Simmons H. A., Sandberg A. L., Canalis E. Direct stimulation of bone resorption by epidermal growth factor. Endocrinology. 1980 Jul;107(1):270–273. doi: 10.1210/endo-107-1-270. [DOI] [PubMed] [Google Scholar]
  26. Shupnik M. A., Ip N. Y., Tashjian A. H., Jr Characterization and regulation of receptors for epidermal growth factor in mouse calvaria. Endocrinology. 1980 Dec;107(6):1738–1746. doi: 10.1210/endo-107-6-1738. [DOI] [PubMed] [Google Scholar]
  27. Shupnik M. A., Tashjian A. H., Jr Epidermal growth factor and phorbol ester actions on human osteosarcoma cells. Characterization of response and nonresponsive cell lines. J Biol Chem. 1982 Oct 25;257(20):12161–12164. [PubMed] [Google Scholar]
  28. Stern P. H., Krieger N. S., Nissenson R. A., Williams R. D., Winkler M. E., Derynck R., Strewler G. J. Human transforming growth factor-alpha stimulates bone resorption in vitro. J Clin Invest. 1985 Nov;76(5):2016–2019. doi: 10.1172/JCI112202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tashjian A. H., Jr, Levine L. Epidermal growth factor stimulates prostaglandin production and bone resorption in cultured mouse calvaria. Biochem Biophys Res Commun. 1978 Dec 14;85(3):966–975. doi: 10.1016/0006-291x(78)90638-1. [DOI] [PubMed] [Google Scholar]
  30. Tashjian A. H., Jr, Voelkel E. F., Lazzaro M., Singer F. R., Roberts A. B., Derynck R., Winkler M. E., Levine L. Alpha and beta human transforming growth factors stimulate prostaglandin production and bone resorption in cultured mouse calvaria. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4535–4538. doi: 10.1073/pnas.82.13.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Westermark K., Karlsson F. A., Westermark B. Epidermal growth factor modulates thyroid growth and function in culture. Endocrinology. 1983 May;112(5):1680–1686. doi: 10.1210/endo-112-5-1680. [DOI] [PubMed] [Google Scholar]
  32. YOUNG C. W., HODAS S. HYDROXYUREA: INHIBITORY EFFECT ON DNA METABOLISM. Science. 1964 Nov 27;146(3648):1172–1174. doi: 10.1126/science.146.3648.1172. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES