Clinical & Experimental Metastasis

, Volume 18, Issue 3, pp 253–260 | Cite as

Expression of bone sialoprotein and osteopontin in breast cancer bone metastases

  • Tharwat Ibrahim
  • Iona Leong
  • Otto Sanchez-Sweatman
  • Rama Khokha
  • Jaro Sodek
  • Howard C. Tenenbaum
  • Bernard Ganss
  • Sela Cheifetz


Bone sialoprotein (BSP) and osteopontin (OPN) are prominent, mineral-associated proteins in the extracellular matrix of bone that have been implicated in the metastatic activity of cancer cells. The expression of BSP, which is normally restricted to mineralizing tissues, has been observed in cancers with a high propensity for forming bone metastases. To investigate the relationship between BSP expression and the formation of bone metastases we have conducted an initial study of the expression of BSP in 10 intraductal breast carcinoma bone metastases using immunostaining and in situ hybridization, and compared the expression with OPN. The metastases were characterized by the infiltration of tumour cells into bone with extensive bone resorption evident. Moderate to strong staining for BSP was observed in all (100%) carcinomas, which also expressed BSP mRNA as determined by in situ hybridization. Variable staining for BSP was also observed in the mineralized bone and expression of BSP mRNA could be observed in osteoblastic cells on the bone surface and in some osteocytes at sites of bone remodelling. Contrary to a previous report, BSP expression could be demonstrated by PCR in three breast cancer cell lines, MCF-7, T47-D and MDA-MB-231. Moreover, in sub-cutaneous tumours formed by MDA-MB-231 breast cancer cells injected into athymic mice, higher immunostaining for BSP was seen in large ulcerating tumours in which mineral deposits were formed. In contrast to BSP, staining for OPN in bone metastases was generally restricted to the interface between tumor cells and bone surface of the carcinomas. While OPN staining was also observed in the cytoplasm of osteoclasts, which showed strong hybridization to a digoxygenin-labelled OPN cRNA probe, expression of OPN was not clearly detectable in the tumour cells. These studies provide the first demonstration of BSP expression by tumour cells in bone metastases and support the concept that BSP may have a role in targeting metastatic cells to bone. Expression of OPN in bone metastases appears to be related to increased bone resorptive activity by osteoclasts.

bone sialoprotein osteopontin breast cancer metastasis bone metastases immunohistochemistry in situ hybridization 


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  1. 1.
    Jacob K, Webber M, Benayahu D et al. Osteonectin promotes prostate cancer cell migration and invasion. A possible mechanism for metastasis to bone. Cancer Res 1999; 59: 4453–7.PubMedGoogle Scholar
  2. 2.
    Brown LF, Papadopoulos-Sergiou A, Berse B et al. Osteopontin expression and distribution in human carcinomas. Am J Path 1994; 145: 610–23.PubMedGoogle Scholar
  3. 3.
    Oates AJ, Barraclough R, Rudland PS. The role of osteopontin in tumorigenesis and metastasis. Invasion & Metastasis 1997; 17: 1–5.Google Scholar
  4. 4.
    Bellahcène A, Castronovo V. Increased expression of osteonectin and osteopontin, two bone matrix proteins, in human breast cancer. Am J Pathol 1995; 146: 95–100.PubMedGoogle Scholar
  5. 5.
    Bellahcène A, Kroll M, Liebens F et al. Bone sialoprotein expression in primary human breast cancer is associated with bone metastases development. J Bone Miner Res 1996; 11: 665–70.PubMedGoogle Scholar
  6. 6.
    Bellahcène A, Maloujahmoum N, Fisher LW et al. Expression of bone sialoprotein in human lung cancer. Calcif Tissue Int 1997; 61: 183–8.PubMedCrossRefGoogle Scholar
  7. 7.
    Hirota S, Ito A, Nagoshi J et al. Expression of bone matrix protein messenger ribonucleic acids in human breast cancers. Possible involvement of osteopontin in development of calcifying foci. Lab Invest 1995; 72: 64–9.PubMedGoogle Scholar
  8. 8.
    Gillespie MT, Thomas RJ, Pu ZY et al. Calcitonoin receptors, bone sialoprotein and osteopontin are expressed in primary breast tumours. Int J Cancer 1997; 73: 812–5.PubMedCrossRefGoogle Scholar
  9. 9.
    Waltregny D, Bellahcène A, Van Riet I et al. Prognostic value of bone sialoprotein expression in clinically localized human prostate cancer. J Natl Cancer Inst 1998; 90: 1000–8.PubMedCrossRefGoogle Scholar
  10. 10.
    Sharp JA, Sung V, Slavin J et al. Tumor cells are the source of osteopontin and bone sialoprotein expression in human breast cancer. Lab Invest 1999; 79: 869–77.PubMedGoogle Scholar
  11. 11.
    Delmas PD, Malaval L. The proteins of bone. In Mundy GR, Martin TJ (eds): Handbook of Experimental Pharmacology, 107: Physiology and Pharmacology of Bone, 1993; 673-724.Google Scholar
  12. 12.
    Ganss B, Kim RH, Sodek J. Bone sialoprotein. Crit Revs Oral Biol & Med 1999; 10: 79–98.Google Scholar
  13. 13.
    Sodek J, Ganss B, McKee MD. Osteopontin. Crit Revs. Oral Biol. & Med., 2000; 11: 279–303.CrossRefGoogle Scholar
  14. 14.
    Rosol TJ. Pathogenesis of bone metastases: Role of tumor-related proteins. J Bone Miner Res 2000; 15: 844–50.PubMedCrossRefGoogle Scholar
  15. 15.
    Oldberg A, Franzén A, Heinegärd D. The primary structure of bone sialoprotein. J Biol Chem 1988; 263: 19430–2.PubMedGoogle Scholar
  16. 16.
    Fisher LW, McBride OW, Termine JD et al. Human bone sialoprotein. Deduced protein sequence and chromosomal localization. J Biol Chem 1990; 265: 2347–51.PubMedGoogle Scholar
  17. 17.
    Chen J, Shapiro H S, Sodek J. Developmental expression of bone sialoprotein mRNA in rat mineralized connective tissues. J Bone Miner Res 1992; 7: 987–97.PubMedGoogle Scholar
  18. 18.
    MacNeil RL, Sheng N, Strayhorn C et al. Bone sialoprotein is localized to the root surface during cementogenesis. J Bone Min Res 1994; 9: 1597–606.Google Scholar
  19. 19.
    Bellahcène A, Merville M-P, Castronovo V. Expression of bone sialoprotein, a bone matrix protein, in human breast cancer. Cancer Res 1994; 54: 2833–26.Google Scholar
  20. 20.
    Bellahcène A, Menard S, Bufalino R et al. Expression of bone sialoprotein in primary human breast cancer is associated with poor survival. Int J Cancer 1996; 69: 350–3.PubMedCrossRefGoogle Scholar
  21. 21.
    Bellahcène A, Antoine N, Clausse N et al. Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels. Lab Invest 1996; 75: 203–10.PubMedGoogle Scholar
  22. 22.
    Castronovo V, Bellahcène A. Evidence that breast cancer associated microcalcifications are mineralized malignant cells. Int J Oncol 1998; 12: 305–8.PubMedGoogle Scholar
  23. 23.
    Withold W, Armbruster FP, Karmatschek M et al., Bone sialoprotein in serum of patients with malignant bone-diseases. Clinical Chem 1997; 43: 85–91.Google Scholar
  24. 24.
    Diel IJ, Solomayer EF, Seibel MJ et al. Serum bone sialoprotein in patients with primary breast cancer is a prognostic marker for subsequent bone metastasis. Clin Cancer Res 1999; 5: 3914–9.PubMedGoogle Scholar
  25. 25.
    Singhal H, Bautista DS, Tonkin KS et al. Elevated plasma osteopontin in metastatic breast cancer associated with increased tumor burden and decreased survival. Clin Cancer Res 1997; 3: 605–11.PubMedGoogle Scholar
  26. 26.
    Tuck AB, O'Malley FP, Singhal H et al. Osteopontin and p53 expression are associated with tumor progression in a case of synchronous, bilateral invasive mammary carcinomas. Arch Pathol Lab Med 1997; 121: 578–84.PubMedGoogle Scholar
  27. 27.
    Denhardt DT, Guo X. Osteopontin: A protein with diverse functions. FASEB J 1993; 7: 1475–82.PubMedGoogle Scholar
  28. 28.
    Denhardt DT, Chambers AF. Overcoming obstacles to metastasis-defenses against host defenses: Osteopontin (OPN) as a shield against attack by cytotoxic host cells. J Cell Biochem 1994; 56: 48–51.PubMedCrossRefGoogle Scholar
  29. 29.
    Crawford HC, Matrisian LM, Liaw L. Distinct roles of osteopontin in host defense activity and tumor survival during squamous cell carcinoma progression in vivo. Cancer Res 1998; 58: 5206–15.PubMedGoogle Scholar
  30. 30.
    Scatena M, Almeida M, Chaisson ML et al. NFkB mediates alphavbeta3 integrin-induced endothelial cell survival. J Cell Biol 1998; 141: 1083–93.PubMedCrossRefGoogle Scholar
  31. 31.
    Fedarko NS, Fohr B, Robey PG et al., Factor H binding to bone sialoprotein and osteopontin enables tumor evasion of complement mediated attack. J Biol Chem 2000; 275: 16666–672.PubMedCrossRefGoogle Scholar
  32. 32.
    Shiraga H, Min W, VanDusen WJ et al. Inihibition of calcium oxalate crystal growth in vitro by uropontin: Another member of the aspartic acid-rich protein superfamily. Proc Natl Acad Sci USA 1992; 89: 426–30.PubMedCrossRefGoogle Scholar
  33. 33.
    Fisher LW, Stubbs JT, Young MF. Antisera and CDNA probes to human and certain animal model bone matrix noncollagenous proteins. Acta Orthop Scand 1995; 66 (Suppl 266): 61–5.Google Scholar
  34. 34.
    Waltregny D, Bellahcène A, de Leval X et al. Increased expression of bone sialoprotein in bone metastases compared to visceral metastases in human breast and prostate cancers. J Bone Miner Res 2000; 15: 834–43.PubMedCrossRefGoogle Scholar
  35. 35.
    Stewart JF, King RGB, Sexton SA et al., Oestrogen receptors, site of metastastic disease and survival in recurrent breast cancer. Eur J Cancer 1981; 17: 449–53.PubMedGoogle Scholar
  36. 36.
    Orr WF, Kostenuik P, Sanchez-Sweatman OH et al. Mechanisms involved in the metastasis of cancer to bone. Breast Cancer Res Treat 1993; 25: 151–63.PubMedCrossRefGoogle Scholar
  37. 37.
    Yoneda T, Sasaki A, Mundy GR. Osteolytic bone disease in breast cancer. Breast Cancer Res Treat 1994; 32: 73–84.PubMedCrossRefGoogle Scholar
  38. 38.
    Sasaguri K, Ganss B, Sodek J et al. Expression of bone sialoprotein in mineralized tissues of tooth and bone and in buccal-pouch carcinomas of Syrian golden hamsters. Arch Oral Biol 2000; 45: 551–62.PubMedCrossRefGoogle Scholar
  39. 39.
    Bosse A, Wuisman P, Jones DB et al. Noncollagenous proteins in heterotopic ossification. Immunohistochemical analysis in 15 paraplegics. Acta Orthop Scand 1993; 64: 634–8.PubMedCrossRefGoogle Scholar
  40. 40.
    McKee MD, Giachelli CM, Nanci A. Matrix-mineral relationships in calcifying human atherosclerotic plaque: Ultrastructural immunodetection of osteopontin and bone sialoprotein at calcification sites. J Bone Miner Res 1996; 11: S330.CrossRefGoogle Scholar
  41. 41.
    Contri MB, Boraldi F, Taparelli F et al. Matrix proteins with high affinity for calcium ions are associated with mineralization within the elastic fibers of pseudoxanthoma elasticum dermis. Am J Pathol 1996; 148: 569–77.PubMedGoogle Scholar
  42. 42.
    Hunter GK, Goldberg HA. Nucleation of hydroxyapatite by bone sialoprotein. Proc Natl Acad Sci USA 1993; 90: 8562–5.PubMedCrossRefGoogle Scholar
  43. 43.
    Oldberg A, Franzèn A, Heinegärd D et al. Identification of a bone sialoprotein receptor in osteosarcoma cells. J Biol Chem 1988; 263: 19433–6.PubMedGoogle Scholar
  44. 44.
    Varner JA, Cheresh DA. Integrins and Cancer. Curr Opin Cell Biol 1996; 8: 724–30.PubMedCrossRefGoogle Scholar
  45. 45.
    Fang F, Orend G, Watanabe N et al. Dependence of cyclin E-CDK2 kinase activity on cell anchorage. Science 1996; 271: 499–502.PubMedGoogle Scholar
  46. 46.
    Ashkar S, Weber GF, Panoutsakopoulou V et al. Eta-1 (osteopontin): an early component of type-1 (cell mediated) immunity. Science 2000; 287: 860–4.PubMedCrossRefGoogle Scholar
  47. 47.
    Hunter GK, Hauschka PV, Poole AR et al. Nucleation and inhibition of hydroxyapatite formation by mineralized tissue proteins. Biochem J 1996; 317 (Pt 1): 59–64.PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Tharwat Ibrahim
    • 1
  • Iona Leong
    • 1
  • Otto Sanchez-Sweatman
    • 2
  • Rama Khokha
    • 2
  • Jaro Sodek
    • 3
  • Howard C. Tenenbaum
    • 1
  • Bernard Ganss
    • 1
  • Sela Cheifetz
    • 1
  1. 1.Medical Research Council Group in Periodontal PhysiologyCanada
  2. 2.Ontario Cancer InstituteTorontoCanada
  3. 3.Department of Biochemistry, Faculties of Dentistry and MedicineUniversity of TorontoCanada

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