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. 1993 Jun;91(6):2791–2795. doi: 10.1172/JCI116521

Herbimycin A, a pp60c-src tyrosine kinase inhibitor, inhibits osteoclastic bone resorption in vitro and hypercalcemia in vivo.

T Yoneda 1, C Lowe 1, C H Lee 1, G Gutierrez 1, M Niewolna 1, P J Williams 1, E Izbicka 1, Y Uehara 1, G R Mundy 1
PMCID: PMC443346  PMID: 8514886

Abstract

Since absence of expression of the c-src gene product in mice indicates that the pp60c-src tyrosine kinase is required and essential for osteoclastic bone resorption, we tested the effects of the antibiotic herbimycin A, which is an inhibitor of pp60c-src on osteoclastic bone resorption in vitro and on hypercalcemia in vivo. We examined the effects of herbimycin A on the formation of bone resorbing osteoclasts in mouse long-term marrow cultures, on isolated rodent osteoclasts and on bone resorption in organ cultures of fetal rat long bones stimulated by parathyroid hormone. We found that herbimycin A in concentrations of 1-100 ng/ml inhibited bone resorption in each of these systems. We determined the effects of herbimycin A (100 ng/ml) on src tyrosine kinase activity in mouse marrow cultures and found that it was decreased. Herbimycin A also decreased elevated blood calcium levels that were induced either by repeated subcutaneous injections of recombinant human interleukin-1 alpha or by a human tumor. There was no evidence for toxicity in any of these culture systems or in mice treated with herbimycin A. A different tyrosine kinase inhibitor that does not inhibit pp60c-src was used as a control and caused none of these effects. These data suggest that pp60c-src tyrosine kinase inhibitors may be useful pharmacologic inhibitors of osteoclastic bone resorption and hypercalcemia.

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Selected References

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

  1. Boyce B. F., Aufdemorte T. B., Garrett I. R., Yates A. J., Mundy G. R. Effects of interleukin-1 on bone turnover in normal mice. Endocrinology. 1989 Sep;125(3):1142–1150. doi: 10.1210/endo-125-3-1142. [DOI] [PubMed] [Google Scholar]
  2. Boyce B. F., Yoneda T., Lowe C., Soriano P., Mundy G. R. Requirement of pp60c-src expression for osteoclasts to form ruffled borders and resorb bone in mice. J Clin Invest. 1992 Oct;90(4):1622–1627. doi: 10.1172/JCI116032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyde A., Ali N. N., Jones S. J. Resorption of dentine by isolated osteoclasts in vitro. Br Dent J. 1984 Mar 24;156(6):216–220. doi: 10.1038/sj.bdj.4805313. [DOI] [PubMed] [Google Scholar]
  4. Li K. W., Geraerts W. P., van Elk R., Joosse J. Quantification of proteins in the subnanogram and nanogram range: comparison of the AuroDye, FerriDye, and India ink staining methods. Anal Biochem. 1989 Oct;182(1):44–47. doi: 10.1016/0003-2697(89)90715-x. [DOI] [PubMed] [Google Scholar]
  5. Li W., Schlessinger J. Platelet-derived growth factor (PDGF)-induced disulfide-linked dimerization of PDGF receptor in living cells. Mol Cell Biol. 1991 Jul;11(7):3756–3761. doi: 10.1128/mcb.11.7.3756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lipsich L. A., Lewis A. J., Brugge J. S. Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses. J Virol. 1983 Nov;48(2):352–360. doi: 10.1128/jvi.48.2.352-360.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lyall R. M., Zilberstein A., Gazit A., Gilon C., Levitzki A., Schlessinger J. Tyrphostins inhibit epidermal growth factor (EGF)-receptor tyrosine kinase activity in living cells and EGF-stimulated cell proliferation. J Biol Chem. 1989 Aug 25;264(24):14503–14509. [PubMed] [Google Scholar]
  8. Oreffo R. O., Bonewald L., Kukita A., Garrett I. R., Seyedin S. M., Rosen D., Mundy G. R. Inhibitory effects of the bone-derived growth factors osteoinductive factor and transforming growth factor-beta on isolated osteoclasts. Endocrinology. 1990 Jun;126(6):3069–3075. doi: 10.1210/endo-126-6-3069. [DOI] [PubMed] [Google Scholar]
  9. Oreffo R. O., Mundy G. R., Seyedin S. M., Bonewald L. F. Activation of the bone-derived latent TGF beta complex by isolated osteoclasts. Biochem Biophys Res Commun. 1989 Feb 15;158(3):817–823. doi: 10.1016/0006-291x(89)92795-2. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Soriano P., Montgomery C., Geske R., Bradley A. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell. 1991 Feb 22;64(4):693–702. doi: 10.1016/0092-8674(91)90499-o. [DOI] [PubMed] [Google Scholar]
  12. Takahashi N., Yamana H., Yoshiki S., Roodman G. D., Mundy G. R., Jones S. J., Boyde A., Suda T. Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. Endocrinology. 1988 Apr;122(4):1373–1382. doi: 10.1210/endo-122-4-1373. [DOI] [PubMed] [Google Scholar]
  13. Thomas J. E., Soriano P., Brugge J. S. Phosphorylation of c-Src on tyrosine 527 by another protein tyrosine kinase. Science. 1991 Oct 25;254(5031):568–571. doi: 10.1126/science.1719633. [DOI] [PubMed] [Google Scholar]
  14. Uehara Y., Fukazawa H. Use and selectivity of herbimycin A as inhibitor of protein-tyrosine kinases. Methods Enzymol. 1991;201:370–379. doi: 10.1016/0076-6879(91)01033-x. [DOI] [PubMed] [Google Scholar]
  15. Yaish P., Gazit A., Gilon C., Levitzki A. Blocking of EGF-dependent cell proliferation by EGF receptor kinase inhibitors. Science. 1988 Nov 11;242(4880):933–935. doi: 10.1126/science.3263702. [DOI] [PubMed] [Google Scholar]
  16. Yoneda T., Alsina M. A., Chavez J. B., Bonewald L., Nishimura R., Mundy G. R. Evidence that tumor necrosis factor plays a pathogenetic role in the paraneoplastic syndromes of cachexia, hypercalcemia, and leukocytosis in a human tumor in nude mice. J Clin Invest. 1991 Mar;87(3):977–985. doi: 10.1172/JCI115106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Yoneda T., Aufdemorte T. B., Nishimura R., Nishikawa N., Sakuda M., Alsina M. M., Chavez J. B., Mundy G. R. Occurrence of hypercalcemia and leukocytosis with cachexia in a human squamous cell carcinoma of the maxilla in athymic nude mice: a novel experimental model of three concomitant paraneoplastic syndromes. J Clin Oncol. 1991 Mar;9(3):468–477. doi: 10.1200/JCO.1991.9.3.468. [DOI] [PubMed] [Google Scholar]
  18. Yoneda T., Lyall R. M., Alsina M. M., Persons P. E., Spada A. P., Levitzki A., Zilberstein A., Mundy G. R. The antiproliferative effects of tyrosine kinase inhibitors tyrphostins on a human squamous cell carcinoma in vitro and in nude mice. Cancer Res. 1991 Aug 15;51(16):4430–4435. [PubMed] [Google Scholar]

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