Skip to main content
SpringerLink
Log in

Mammary tumor development in MMTV-c-myc/MMTV-v-Ha-ras transgenic mice is unaffected by osteopontin deficiency

  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Transgenic mice expressing c-myc and v-Ha-ras specifically in the mammary gland under the control of the mammary specific promoter MMTV develop unifocal mammary tumors with a half time of about 46 days, and these tumors express high levels of osteopontin mRNA and protein. In order to evaluate the requirement for osteopontin expression by these tumors, we have crossed transgenic mice expressing these two oncogenes with mice with a targeted disruption of the osteopontin gene. Littermates expressing both myc and ras, and with either wild-type or disrupted OPN alleles were evaluated for tumor incidence and growth rate. Both of these parameters were found to be unaffected by a lack of osteopontin in the whole animal. Ras and myc expression level, measured at the level of mRNA, was not different in tumors of the two genotypes. Macrophage accumulation, while extremely variable among different tumors, did not correlate with the OPN status of the animals. Expression of the related gene BSP was not detected in any of the tumors, and was similar in bones of wildtype and OPN –/– mice. Similarly, the vitronectin gene was expressed at very low levels in tumors of either genotype. These results indicate that despite its high level of expression, OPN is either not required for mammary primary tumor formation and growth in this system, or can be replaced by molecules other than BSP and vitronectin in mice that totally lack osteopontin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Denhardt DT, Noda M: Osteopontin expression and function: role in bone remodeling. J Cell Biochem S30/31, 92-102, 1998

    Google Scholar 

  2. Smith LL, Cheung HK, Ling LE, Chen J, Sheppard D, Pytela R, Giachelli CM: Osteopontin N-terminal domain contains a cryptic adhesive sequence recognized by a9b1 integrin. J Biol Chem 271: 28485-28491, 1996

    Google Scholar 

  3. Yokosaki Y, Matsuura N, Sasaki T, Murakami I, Schneider H, Higashiyama S, Saitoh Y, Yamakido M, Taooka Y, Sheppard D: The integrin alpha(9)beta(1) binds to a novel recognition sequence (SVVYGLR) in the thrombin-cleaved amino-terminal fragment of osteopontin. J Biol Chem 274: 36328-36334, 1999

    Google Scholar 

  4. van Dijk S, D'Errico JA, Somerman MJ, Farach-Carson MC, Butler WT: Evidence that a non-RGD domain in rat osteopontin is involved in cell attachment. J Bone Miner Res 8: 1499-1506, 1993

    Google Scholar 

  5. Katagiri YU, Murakami M, Mori K, Iizuka J, Hara T, Tanaka K, Jia WY, Chambers AF, Uede T: Non-RGD domains of osteopontin promote cell adhesion without involving alpha v integrins. J Cell Biochem 62: 123-131, 1996

    Google Scholar 

  6. Katagiri YU, Sleeman J, Fujii H, Herrlich P, Hotta H, Tanaka K, Chikuma S, Yagita H, Okumura K, Murakami M, Saiki I, Chambers AF, Uede T: CD44 variants but not CD44s cooperate with beta1-containing integrins to permit cells to bind to osteopontin independently of arginine-glycine-aspartic acid, thereby stimulating cell motility and chemotaxis. Cancer Res 59: 219-226, 1999

    Google Scholar 

  7. Weber GF, Ashkar S, Glimcher MJ, Cantor H. Receptor-ligand interaction between CD44 and osteopontin (ETA-1). Science 271: 509-512, 1996

    Google Scholar 

  8. Butler WT, Ridall AL, McKee MD: Osteopontin. In: Bilezikian JP, Raisz LG, Rodan GA, (eds) Principles in bone biology. San Diego: Academic Press, 1996, p. 167-181

    Google Scholar 

  9. Rittling SR, Matsumoto HN, McKee MD, Nanci A, An XR, Novick KE, Kowalski AJ, Noda M, Denhardt DT: Mice lacking osteopontin show normal development and bone structure but display altered osteoclast formation in vitro. J Bone Min Res 13: 1101-1111, 1998

    Google Scholar 

  10. Liaw L, Birk DE, Ballas CB, Whitsitt JS, Davidson JM, Hogan BLM: Altered wound healing in mice lacking a functional osteopontin gene (spp1). J Clin Invest 101: 1468-1478, 1998

    Google Scholar 

  11. Rittling SR, Denhardt DT: Osteopontin (OPN) function in pathology: lessons from OPN-deficient mice. Experimental Nephrology 7: 103-113, 1999

    Google Scholar 

  12. Senger DR, Perruzzi CA, Gracey CF, Papadopoulous A, Tenen DG: Secreted phosphoproteins associated with neoplastic transformation: close homology with plasma proteins cleaved during blood coagulation. Cancer Res 48: 5770-5774, 1988

    Google Scholar 

  13. Senger DR, Perruzzi CA, Papadopoulous A: Elevated expression of secreted phosphoprotein I (osteopontin, 2ar) as a consequence of neoplastic transformation. Anticancer Res 9: 1291-1300, 1989

    Google Scholar 

  14. Chambers AF, Behrend EI, Wilson SM, Denhardt DT: Induction of expression of osteopontin (OPN, secreted phosphoprotein) in metastatic, ras-transformed NIH-3T3 cells. Anticancer Res 12: 43-48, 1992

    Google Scholar 

  15. Guo X, Zhang YP, Mitchell DA, Denhardt DT, Chambers AF: Identification of a ras-activated enhancer in the mouse osteopontin promoter and its interaction with a putative ETSrelated transcription factor whose activity correlates with the metastatic potential of the cell. Mol Cell Bio 15: 476-487, 1995

    Google Scholar 

  16. Morris VL, Tuck AB, Wilson SM, Percy D, Chambers AF: Tumor progression and metastasis in murine D2 hyperplastic alveolar nodule mammary tumor cell lines. Clin Exp Metastasis 11: 103-112, 1993

    Google Scholar 

  17. Behrend EI, Craig AM, Wilson SM, Denhardt DT, Chambers AF: Reduced malignancy of ras-transformed NIH 3T3 cells expressing antisense osteopontin RNA. Cancer Res 54: 832-837, 1994

    Google Scholar 

  18. Gardner HAR, Berse B, Senger DR: Specific reduction in osteopontin synthesis by antisense RNA inhibits the tumorigenicity of transformed Rat1 fibroblasts. Oncogene 9: 2321-2326, 1994

    Google Scholar 

  19. Su L, Mukherjee AB, Mukherjee BB: Expression of antisense osteopontin mRNA inhibits tumor promoter-induced neoplastic transformation of mouse JB6 epidermal cells. Oncogene 10: 2163-2169, 1995

    Google Scholar 

  20. Feng B, Rollo EE, Denhardt DT: Osteopontin (OPN) may facilitate metastasis by protecting cells from macrophage NO-mediated cytotoxicity: evidence from cell lines downregulated for OPN expression. Clin Exptl Metastasis 13: 453-462, 1995

    Google Scholar 

  21. Oates AJ, Barraclough R, Rudland PS: Enhanced expression of the osteopontin gene in metastatic rat mammary epithelialderived cell lines. Oncogene 97-104, 1996

  22. Chen H, Ke Y, Oates AJ, Barraclough R, Rudland PS: Isolation of and effector for metastasis-inducing DNAs from a human metastatic carcinoma cell line. Oncogene 14: 1581-1588, 1997

    Google Scholar 

  23. Yoneda T, Williams PJ, Niewolna M: Promotion of angiogenesis and enhancement of breast cancer metastasis to bone. Bone 23: S201, 1998

    Google Scholar 

  24. Rittling SR, Novick KW: Osteopontin expression in mammary gland development and tumorigenesis. Cell Growth Differ 8: 1061-1069, 1997

    Google Scholar 

  25. Brown LF, Papadopoulous-Sergiou A, Berse B, Manseau EJ, Tognazzi K, Perruzzi CA, Dvorak HF, Senger DR: Osteopontin expression and distribution in human carcinomas. Am J Pathol 145: 610-623, 1994

    Google Scholar 

  26. Sung V, Gilles C, Murray A, Clarke R, Aaron AD, Azumi N, Thompson EW: The LCC-15 MB human breast cancer cell line expresses osteopontin and exhibits an invasive and metastatic phenotype. Exptl Cell Res 241: 273-284, 1998

    Google Scholar 

  27. Tuck AB, O'Malley FP, Singhal H, Tonkin KS, Kerkvliet N, Saad Z, Doig GS, Chambers AF: Osteopontin expression in a group of lymph node negative breast cancer patients. Int J Cancer 79: 502-508, 1998

    Google Scholar 

  28. Sinn E, Muller WJ, Pattengale PK, Tepler I, Wallace R, Leder P: Coexpression ofMMTV/v-Ha-ras andMMTV-c-myc genes in transgenic mice: synergistic action of oncogenes in vivo. Cell 49: 465-475, 1987

    Google Scholar 

  29. Pattengale PK, Stewart TA, Leder A, Sinn E, Tepler I, Schmidt E, Leder P: Animal models of human disease. Am J Pathol 135: 39-61, 1989

    Google Scholar 

  30. Young MF, Ibaraki K, Kerr JM, Lyu MS, Kozak CA: Murine bone sialoprotein (BSP): cDNA cloning, mRNA expression and genetic mapping. Mamm Genome 5: 108-111, 1994

    Google Scholar 

  31. Rittling SR, Feng F: Detection of mouse osteopontin by western blotting. Biochem Biophys Res Comm 250: 287-292, 1998

    Google Scholar 

  32. Liaw L, Almeida M, Hart CE, Schwartz SM, Giachelli CM: Osteopontin promotes vascular cell adhesion and spreading and is chemotactic for smooth muscle cells in vitro. Circ Res 74: 214-224, 1994

    Google Scholar 

  33. Austyn JM, Gordon S: F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol 11: 805-815, 1981

    Google Scholar 

  34. Singh RP, Patarca R, Schwartz J, Singh P, Cantor H: Definition of a specific interaction between the early T lymphocyte activation 1 (Eta-1) protein and murine macrophages in vitro and its effect upon macrophages in vivo. J Exp Med 171: 1931-1942, 1990

    Google Scholar 

  35. Giachelli CM, Lombardi D, Johnson RJ, Murry CE, Almeida M: Evidence for a role of osteopontin in macrophage infiltration in response to pathological stimuli in vivo. Am J Pathol 152: 353-358, 1998

    Google Scholar 

  36. Fisher LW, McBride OW, Termine JD, Young MF: Human bone sialoprotein. Deduced protein sequence and chromosomal localization. J Biol Chem 265: 2347-2351, 1990

    Google Scholar 

  37. Bellahcene A, Antoine N, Clausse N, Tagliabue E, Fisher LW, Kerr JM, Jares P, Castronovo V: Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels. Lab Invest 75: 203-210, 1996

    Google Scholar 

  38. Sharp JA, Sung V, Slavin J, Thompson EW, Henderson MA: Tumor cells are the source of osteopontin and bone sialoprotein expression in human breast cancer. Lab Invest 79: 869-877, 1999

    Google Scholar 

  39. Nip J, Shibata H, Loskutoff DJ, Cheresh DA, Brodt P: Human melanoma cells derived from lymphatic metastases use integrin alpha v beta 3 to adhere to lymph node vitronectin. J Clin Invest 90: 1406-1413, 1992

    Google Scholar 

  40. 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 58: 5206-5215, 1998

    Google Scholar 

  41. Talts JF, Wirl G, Dictor M, Muller WJ, Fassler R: Tenascin-C modulates tumor stroma and monocyte/macrophage recruitment but not tumor growth or metastasis in a mouse strain with spontaneous mammary cancer. J Cell Sci 112 (Pt 12): 1855-1864, 1999

    Google Scholar 

  42. Wu YM, Denhardt DT, Rittling SR: Osteopontin is required for full expression of the transformed phenotype by the ras oncogene. British J Cancer 83: 156-163, 2000

    Google Scholar 

  43. Hynes RO: Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69: 11-25, 1992

    Google Scholar 

  44. Tuck AB, Arsenault DM, O'Malley FP, Hota C, Ling MC, Wilson SM, Chambers AF: Osteopontin induces increased invasiveness and plasminogen activator expression of human mammary epithelial cells. Oncogene 18: 4237-4246, 1999

    Google Scholar 

  45. Mukaida N, Harada A, Matsushima K: Interleukin-8 (IL-8) and monocyte chemotactic and activating factor (MCAF/MCP-1), chemokines essentially involved in inflammatory and immune reactions. Cytokine Growth Factor Rev 9: 9-23, 1998

    Google Scholar 

  46. Pan H, Yin C, Dyson NJ, Harlow E, Yamasaki L, Van Dyke T: Key roles for E2F1 in signaling p53-dependent apoptosis and in cell division within developing tumors. Mol Cell 2: 283-292, 1998

    Google Scholar 

  47. Crosby AH, Lyu MS, Lin K, McBride OW, Kerr JM, Aplin HM, Fisher LW, Young MF, Kozak CA, Dixon MJ: Mapping of the human and mouse bone sialoprotein and osteopontin loci. Mamm Genome 7: 149-151, 1996

    Google Scholar 

  48. Bianco P, Fisher LW, Young MF, Termine JD, Robey PG: Expression of bone sialoprotein (BSP) in developing human tissues. Calcif Tissue Int 49: 421-426, 1991

    Google Scholar 

  49. Schvartz I, Seger D, Shaltiel S. Vitronectin: Int J Biochem Cell Biol 31: 539-544, 1999

    Google Scholar 

  50. Seiffert D: Constitutive and regulated expression of vitronectin. Histol Histopathol 12: 787-797, 1997

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ, USA

    Fei Feng & Susan R. Rittling

Authors
  1. Fei Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susan R. Rittling
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Feng, F., Rittling, S.R. Mammary tumor development in MMTV-c-myc/MMTV-v-Ha-ras transgenic mice is unaffected by osteopontin deficiency. Breast Cancer Res Treat 63, 71–79 (2000). https://doi.org/10.1023/A:1006466516192

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006466516192

Search

Navigation