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In vitro bone resorption activity produced by a hypercalcemic adenocarcinoma tumor line (CAC-8) in nude mice

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Summary

Tumor extracts and conditioned tissue culture media from a canine adenocarcinoma tumor line (CAC-8) propagated in nude mice significantly increasedin vitro bone resorption in neonatal mouse calvaria as measured by release of previously incorporated45Ca.In vitro bone resorption activity was induced in a dose-dependent manner, was not suppressible by indomethacin, and was heat- and acid-stable. Gel exclusion chromatography demonstrated peak bone resorbing activity at a relative molecular mass of approximately 28,000. The parathyroid hormone (PTH) antagonist (8,18norleucine,34tyrosine) bovine PTH (3–34) amide did not inhibit CAC 8-stimulated or (1–34) bPTH-induced bone resorption. There was an increased number of tartrate-resistant, acid phosphatase-positive cells in calvariae exposed to CAC-8 extract. Ultrastructural evaluation of calvaria revealed hypertrophy and maturation of osteoclasts in calvaria exposed to CAC-8 extract. The maturation effects included close contact to bone surfaces and the presence of clear zones and ruffled borders in osteoclasts. Similar structures were observed infrequently in osteoclasts of control calvaria. These data demonstrate that the tumor line (CAC-8) contained activity capable of stimulatingin vitro bone resorption by increasing osteoclast numbers and the activity of existing osteoclasts.

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References

  1. Mundy GR, Ibbotson KJ, D'Souza SM, Simpson EL, Jacobs JW, Martin TJ (1984) The hypercalcemia of cancer. Clinical implications and pathogenic mechanisms. N Engl J Med 310:1718–1727

    Article  PubMed  CAS  Google Scholar 

  2. Mundy GR, Ibbotson KJ, D'Souza SM (1985) Tumor products and the hypercalcemia of malignancy. J Clin Invest 76:391–394

    PubMed  CAS  Google Scholar 

  3. Mundy GR, Jacobs JW, Ibbotson KJ, D'Souza SM, Bertolini DR, Simpson EL (1984) Hypercalcemia of malignancy. In: Cohn DV, Fujita T, Potts JT Jr, Talmage RV (eds) Endocrine control of bone and calcium metabolism. Elsevier Science Publications, Amsterdam, pp 278–283

    Google Scholar 

  4. Meuten DJ, Segre GV, Capen CC, Kociba GJ, Voekel EF, Levine L, Tashjian AH Jr, Chew DJ, Nagode LA (1983) Hypercalcemia in dogs with adenocarcinoma derived from apocrine glands of the anal sac. Biochemical and histomorphometric investigations. Lab Invest 48:428–435

    PubMed  CAS  Google Scholar 

  5. Rosol TJ, Capen CC, Weisbrode SE, Horst RL (1986) Humoral hypercalcemia of malignancy: nude mouse model of a canine adenocarcinoma derived from apocrine glands of the anal sac. Biochemical, histomorphometric and ultrastructural studies. Lab Invest 54:679–688

    PubMed  CAS  Google Scholar 

  6. Roberts AB, Lamb LC, Newton DL, Sport MB, DeLarco JE, Todaro GJ (1980) Transforming growth factors: isolation of polypeptides from virally and chemically transformed cells by acid/ethanol extraction. Cell Biology 77:3494–3498

    CAS  Google Scholar 

  7. Lau KHW, Lee MY, Linkhart TA, Mohan S, Vermeiden J, Lui CC, Baylink DJ (1985) A mouse tumor-derived osteolytic factor stimulates bone resorption by a mechanism involving local prostaglandin production in bone. Biochim Biophys Acta 840:56–68

    PubMed  CAS  Google Scholar 

  8. Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  9. Minkin C, Fredericks RS, Pokress S, Rude RK, Sharp CF Jr, Tong M, Singer FR (1981) Bone resorption and humoral hypercalcemia of malignancy: stimulation of bone resorptionin vitro by tumor extracts is inhibited by prostaglandin synthesis inhibitors. J Clin Endocrinol Metab 53:941–947

    PubMed  CAS  Google Scholar 

  10. Brinn NT, Pickett JP (1979) Glycol methacrylate for routine special stains, histochemistry, enzyme histochemistry and immunochemistry. J Histotechnol 2:125–130

    CAS  Google Scholar 

  11. Minkin C (1982) Bone acid phosphatase: tartrate-resistant acid phosphatase as a marker of osteoclast function. Calcif Tissue Int 34:285–290

    Article  PubMed  CAS  Google Scholar 

  12. Holtrop ME (1976) Quantification of the ultrastructure of the osteoclast for the evaluation of cell function. In: Meunier PJ (ed) Proceedings of the Second International Workshop on Bone Histomorphometry. Armour Montagu, Paris, pp 133–145

    Google Scholar 

  13. Ibbotson KJ, D'Souza SM, Ng KW, Osborne CK, Niall M, Martin TJ, Mundy GR (1983) Tumor-derived growth factor increases bone resorption in a tumor associated with humoral hypercalcemia of malignancy. Science 221:1292–1294

    Article  PubMed  CAS  Google Scholar 

  14. D'Souza SM, Ibbotson KJ, Smith DD, Mundy GR (1983) Co-purification of a transforming growth factor (TGF) with bone resorbing activity in the Walker rat carcinosarcoma: a mechanism for the humoral hypercalcemia of malignancy (HHM). Calcif Tissue Int 35:658A

    Google Scholar 

  15. Ibbotson KJ, D'Souza SM, Smith DD, Carpenter G, Mundy GR (1985) EGE receptor antiscrum inhibits bone resorbing activity produced by a rat Leydig cell tumor associated with the humoral hypercalcemia of malignancy. Endocrinology 116:469–471

    Article  PubMed  CAS  Google Scholar 

  16. Raigz LG, Simmons HA, Sandberg AL, Canalis E (1980) Direct stimulation of bone resorption by epidermal growth factor. Endocrinology 107:270–273

    Google Scholar 

  17. Tashjian AH Jr, Voelkel EF, Lazzaro M, Singer FR, Roberts AB, Derynck R, Winkler ME, Levine L (1985) α and β human transforming growth factors stimulate prostaglandin production and bone resorption in cultured mouse calvaria. Proc Natl Acad Sci USA 82:4535–4538

    Article  PubMed  CAS  Google Scholar 

  18. Stern PH, Krieger NS, Nissenson RA, Williams RD, Winkler ME, Derynck R, Strewler GJ (1985) Human transforming growth factor-alpha stimulates bone resorption in vitro. J Clin Invest 76:2016–2019

    PubMed  CAS  Google Scholar 

  19. Rosol TJ, Capen CC (1986) In vitro bone resorption and transforming growth factor (TGF) activities in hypercalcemic canine adenocarcinoma tumor-line (CAC-8) maintained in nude mice. J Bone Mineral Res 1(Suppl 1):180

    Google Scholar 

  20. Dewhirst FE, Stashenko PP, Mole JE, Tsurumachi T (1985) Purification and partial sequence of human-osteoclast activating factor: identity with interleukin 1β. J Immunol 135:2562–2568

    PubMed  CAS  Google Scholar 

  21. D'Souza SM, Ibbotson KJ, Mundy GR (1984) Failure of parathyroid hormone antagonists to inhibit in vitro bone resorbing activity produced by two animal models of the humoral hypercalcemia of malignancy. J Clin Invest 74:1104–1107

    PubMed  Google Scholar 

  22. Klein RF, Strewler GJ, Nissenson RA, Williams RD, Leung SC (1985) PTH-like protein from human carcinoma cells stimulates bone resorption. J Bone Min Res 1:96

    Google Scholar 

  23. Rodan SB, Insogna KL, Vignery AMC, Stewart AF, Broadus AE, D'Souza SM, Bertolini DR, Mundy GR, Rodan GA (1983) Factors associated with humoral hypercalcemia stimulate adenylate cyclase in osteoblastic cells. J Clin Invest 72:1511–1515

    Article  PubMed  CAS  Google Scholar 

  24. Rosol TJ, Capen CC, Brooks CL (in press) Bone and kidney adenylate cyclase-stimulating activity produced by a hypercalcemic canine adenocarcinoma line (CAC-8) maintained in nude mice. Cancer Res.

  25. Holtrop ME, Raisz LG (1979) Comparison of the effects of 1,25-dihydroxycholecalciferol, prostaglandin E2, and osteoclast activating factor with parathyroid hormone on the ultrastructure of osteoclasts in cultured long bones of fetal rats. Calcif Tissue Int. 29:201–205

    Article  PubMed  CAS  Google Scholar 

  26. Schelling SH, Wolfe HJ, Tashjian AH Jr (1980) Role of the osteoclast in prostaglandin E2-stimulated bone resorption: a correlative morphometric and biochemical analysis. Lab Invest 42:290–295

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Veterinary Pathobiology, The Ohio State University, 925 Coffey Road, 43210, Columbus, Ohio, USA

    T. J. Rosol &  C. C. Capen

  2. Department of Basic Sciences, University of Southern California School of Dentistry, 90007, Los Angeles, California, USA

    C. Minkin

Authors
  1. T. J. Rosol
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  2. C. C. Capen
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  3. C. Minkin
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Rosol, T.J., Capen, C.C. & Minkin, C. In vitro bone resorption activity produced by a hypercalcemic adenocarcinoma tumor line (CAC-8) in nude mice. Calcif Tissue Int 39, 334–341 (1986). https://doi.org/10.1007/BF02555201

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  • DOI: https://doi.org/10.1007/BF02555201

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