Skip to main content
SpringerLink
Log in

Epidermal growth factor in NMU-induced mammary tumors in rats

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

Abstract

In this work we analyze the hypothesis that tumors induced by ip N-nitroso-N-methylurea injection express EGF-like peptides and EGF receptors which could be involved in the response to hormone manipulation. EGF receptors (EGFR) were determined in the purified membrane fraction of tumors from control and ovariectomized (OVX) animals and no significant differences were found in either maximal binding capacities (Q) or dissociation constants (Kd) between them. Neither did we observe differences between tumors that regressed (HR) or continued growing (HU) after ovariectomy. In order to test the ability of EGFR to trigger a biological response we measured the production of second messengers inositol triphosphates (IP3) and cAMP levels; we found that EGF increases IP3 production in a dose-dependent way, while cAMP levels were not affected. In addition, EGF was able to induce in vitro cell proliferation in a concentration-dependent manner when tested in primary cultures of tumor cells by the clonogenic soft agar technique. EGF/TGF-α activity was determined by a radioreceptor assay in tumor cytosols from control and OVX rats. Results showed a trend to lower values in tumors from OVX rats, but no differences between HR and HU tumors. A positive correlation was found between EGF/TGF-α activity and progesterone receptor maximal binding capacity. When we tested the action of estradiol and EGF added together to primary cultures of tumor cells we found an additive effect on cell proliferation. The study of steady state mRNA levels showed that E2 increases PgR and c-myc mRNA levels in HR but not in HU tumors. In conclusion, the autocrine loop EGFR-EGF/TGF-α present in all tumors is hormonally regulated, possibly by Pg, but is not related to the tumor response to ovariectomy.

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. Osborne CK, Hamilton B, Titus G, Livingston B: Epidermal growth factor stimulation of human breast cancer cells in culture. Cancer Res 40: 2361-2366, 1980

    Google Scholar 

  2. Fitzpatrick SL, Lachance MP, Schultz GS: Characterization of epidermal growth receptor and action of human breast cancer cells in culture. Cancer Res 44: 3443-3447, 1984

    Google Scholar 

  3. Dickson RB, Huff KK, Spencer EM, Lippman ME: Induction of epidermal growth factor-related polypeptides by 17?-estradiol in MCF-7 cells. Endocrinology 188: 138-143, 1986

    Google Scholar 

  4. Bronzert DA, Pantazis P, Antonaides HN, Kasid A, Davidson N, Dickson RB, Lippman ME: Synthesis and secretion of PDGF-like growth factor for human breast cancer. Proc Natl Acad Sci USA 84: 5763-5767, 1987

    Google Scholar 

  5. Huff KK, Kaufman D, Gabbay KH, Spencer EM, Lippman ME, Dickson RB: Secretion of an insulin-like growth factor - I - related protein by human breast cancer cells. Cancer Res 45: 4613-4619, 1986

    Google Scholar 

  6. Knabbe C, Lippman ME, Wakefield LM, Flanders KC, Kassid A, Derysik R, Dickson RB: Evidence that transforming growth factor-??is a hormonally regulated negative growth factor in human breast cancer cells. Cell 48: 417-428, 1987

    Google Scholar 

  7. Keski-oja J, Postlethwaite AE, Moses HL: Transforming growth factors in the regulation of malignant cell growth and invasion. Cancer Invest 6: 705-724, 1988

    Google Scholar 

  8. Knabbe C, Zugmaier G, Schmahl M, Dietel M, Lippman ME, Dickson RB: Induction of transforming growth factor ??by the antiestrogens droloxifene, tamoxifen and toremifene in MCF-7 cells. Am J Clin Oncol 14: 515-520, 1991

    Google Scholar 

  9. Waterfield MD: Epidermal growth factor and related molecules. Lancet 1: 1243-1246, 1989

    Google Scholar 

  10. Singletary V, Baker FL, Spitzer G, Tucker SL, Tomasovic B, Brock WA, Ajani JA, Kelly AM: Biologic effect of epidermal growth factor in the in vitrogrowth of human tumors. Cancer Res 47: 403-406, 1987

    Google Scholar 

  11. Ennis BW, Valverius EM, Bates SE, Lippman ME, Bellot F, Kris R, Schlessinger J, Masui H, Goldember A, Mendelsohn J, Dickson RB: Antiepidermal growth factor receptor antibodies inhibit the autocrine stimulated growth of MDA-486 human breast cancer cells. Mol Endocrinol 3: 1830-1838, 1989

    Google Scholar 

  12. Dickson RB, Bates SE, McManaway ME, Lippman ME: Characterization of estrogen responsive transforming activity in human breast cancer cell lines. Cancer Res 46: 1707-1713, 1986

    Google Scholar 

  13. Bates SE, Valverius EM, Ennis BW, Bronzert DA, Sheridan JP, Stampfer MR, Mendelsohn J, Lippman ME, Dickson RB: Expression of transforming growth factor ?/epidermal growth factor receptor pathway in normal human breast epithelial cells. Endocrinology 126: 596-607, 1990

    Google Scholar 

  14. Manni A, Wright C, Badger B, Bartholomew M, Herlyn M, Mendelsohn J, Masui H, Demers L: Role of transforming growth factor-?-related peptides in the autocrine-paracrine control of experimental breast cancer growth in vitroby estradiol, prolactin and progesterone. Breast Cancer Res Treat 15: 73-83, 1990

    Google Scholar 

  15. Ahmed SR, Badger B, Wright C, Manni A: Role of transforming growth factor-?-(TGF-?) and basal and hormonalstimulated growth by estradiol, prolactin and progesterone in human and rat mammary tumor cells: studies using TGF-??and EGF receptor antibodies. J Steroid Biochem Mol Biol 38: 687-693, 1991

    Google Scholar 

  16. Rivera ES, Andrade N, Martin G, Melito G, Cricco G, Mohamad N, Davio C, Caro R, Bergoc RM: Induction of mammary tumors in rat by intraperitoneal injection of NMU: histopathology and estral cycle influence. Cancer Lett 86: 223-228, 1994

    Google Scholar 

  17. Martin G, Davio C, Rivera E, Melito G, Cricco G, Andrade N, Caro R, Bergoc RM: Hormone dependence of mammary tumors induced in rat by intraperitoneal NMU-injection. Cancer Invest 15(1): 8-17, 1997

    Google Scholar 

  18. Gullino PM, Pettigrew HM, Grantan FH: N-nitrosomethylurea as mammary gland carcinogen in rats. J Natl Cancer Inst 54: 401-409, 1975

    Google Scholar 

  19. Turkot-Lemay L, Kelly PA: Characterization of estradiol, progesterone and prolactin receptors in nitrosomethylureainduced mammary tumors and effect of antiestrogenic treatment on the development and growth of these tumors. Cancer Res 40: 3232-3240, 1980

    Google Scholar 

  20. Rose RB, Pruit B, Stauber P et al.: Influence of dosage schedule on the biological characteristics of N-nitrosomethylurea-induced rat mammary tumors. Cancer Res 40: 235-239, 1980

    Google Scholar 

  21. Welsch CW, Brown HM, Goodrich-Smith M et al.: Synergistic effect of chronic prolactin suppression and retinoic treatment in the prophylaxis of N-methyl-nitrosourea-induced mammary tumorigenesis in female Sprague-Dawley rats. Cancer Res 40: 3095-3098, 1980

    Google Scholar 

  22. Arafah BM, Finegas HM, Roe J et al.: Hormone dependency in N-nitrosomethylurea-induced rat mammary tumors. Endocrinology 111: 584-588, 1982

    Google Scholar 

  23. Martin G, Rivera ES, Davio C, Cricco C, Levin E, Cocca C, Andrade N, Caro R, Bergoc RM: Receptors characterization of intraperitoneally NMU-induced mammary tumors in rat. Cancer Lett 101: 1-8, 1996

    Google Scholar 

  24. Korenman SG, Dukes BA: Specific estrogen binding by the cytoplasm of human breast carcinoma. J Clin Endocrinol Metab 30: 639-645, 1970

    Google Scholar 

  25. Venturino A, Rivera ES, Bergoc RM, Caro R: A simplified competition data analysis for radioligand specific activity determination. Nucl Med Biol 17: 233-237, 1988

    Google Scholar 

  26. Cricco GP, Davio CA, Bergoc RM, Rivera ES: Inhibition of tumor growth induced by histamine: In vivoand in vitrostudies. Agents and Actions 38: Special Conference Issue C175-C177, 1993

  27. Freter CE, Lippman ME, Cheville A, Zinn S, Gelmann EP: Alterations in phosphoinositide metabolism associated with 17?-estradiol and growth factor treatment of MCF-7 breast cancer cells. Mol Endocrinol 2: 159-166, 1988

    Google Scholar 

  28. Rivera ES, Davio CA, Venturino A, Caro RA, Bergoc RM: Histamine receptors in an experimental mammary carcinoma. Biomed Pharmacotherap 48: 399-406, 1994

    Google Scholar 

  29. Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156-159, 1987

    Google Scholar 

  30. Agresti A: In: John Wiley (ed) Categorical Data Analysis, New York, 1990

  31. Dickson RB, Lippman ME: Estrogenic regulation of growth and polypeptide growth factor secretion in human breast carcinoma. Endocrine Rev 8: 29-40, 1987

    Google Scholar 

  32. Ferning DG, Smith JA, Rudland PS: Relationship of growth factors and differentiation in normal and neoplastic development of the mammary gland. In: Lippman M, Dickson R (eds) Regulatory mechanisms in breast cancer, Boston, 1991

  33. Bates SE, Davidson EN, Valverius EM: Expressions of transforming growth factor and its messenger ribonucleic acid in human breast cancer: its regulation by estrogen and its possible functional significance. Mol Endo 2: 543-551, 1988

    Google Scholar 

  34. Boonstra J, Rijken P, Humbel B, Cremers F, Verkleij A, van Bergen Henegouwen P: The epidermal growth factor. Cell Biol Internatl 19: 413-430, 1995

    Google Scholar 

  35. Schlessinger J, Bar-Sagi D: Activation of Ras and other signaling pathways by receptor tyrosine kinases. Cold Spring Harbor Symposia on Quantitative Biology 59: 173-179, 1995

    Google Scholar 

  36. Ullrich A, Schlessinger J: Signal transduction by receptors with tyrosine kinase activity. Cell 61: 203-212, 1990

    Google Scholar 

  37. Soler C, Alvarez CV, Beguinot L, Carpenter G: Potent SHC tyrosine phosphorylation by epidermal growth factor at low receptor density or in the absence of receptor autophosphorylation sites. Oncogene 9: 2207-2215, 1994

    Google Scholar 

  38. Stryjek-Kaminka D, Piiper A, Zeuzem S: Epidermal growth factor regulates adenylate cyclase activity via Gs and Gi 1-2 proteins in pancreatic acinar membranes. Biochem J 316: 87-91, 1996

    Google Scholar 

  39. Stryjek-Kaminka D, Piiper A, Zeuzem S: EGF inhibits secretagogue-induced cAMP production and amylase secretion by Gi proteins in pancreatic acini. Am J Physiol 269(5Pt 1): G676-G682, 1995

    Google Scholar 

  40. Grimaux M, Magdelenat H, Martin PM: Prognosis value of EGF receptors (EGFR) in breast cancer. Study of a group of node positive patients. In: Bolla M, Chambaz EM, Vrousos C (eds) Growth Factors and Oncogenes. Collogne INSERM/ John Libbey Eurotext Ltd. Vol 190, 1989, pp 119-124

  41. Toi M, Nakamura T, Mukarida H, Wada T, Osaki A, Yamada H, Toge T, Niimoto M, Hattori T: Relationship between epidermal growth factor receptor status and various prognostic factors in human breast cancer. Cancer 65: 1980-1984, 1990

    Google Scholar 

  42. Wrba F, Reiner A, Ritzinger E, Holzner JH: Expression of epidermal growth factor receptors (EGF) on breast carcinomas in relation to growth fractions, estrogen receptor status and morphological criteria. An immunohistochemical study. Path Res Pract 183: 25-29, 1988

    Google Scholar 

  43. Koenders PG, Beex LVAM, Kienhuis CBM, Kloppenborg PWC, Benraad ThJ: Epidermal growth factor receptor and prognosis in human breast cancer: a prospective study. Breast Cancer Res Treat 25: 21-27, 1993

    Google Scholar 

  44. Rivera ES, Actis AM, Martin G, Bergoc RM, Levin E: Factores de crecimiento y dependencia hormonal en cáncer de mama. Acta Bioq Clin Latinoamer 26: 11-25, 1992

    Google Scholar 

  45. Barker S, Vinson GP: Epidermal growth factor in breast cancer. Int J Biochem 22: 939-945, 1990

    Google Scholar 

  46. Chrysogelos SA, Yarden RI, Lauber AH, Murphy JM: Mechanisms of EGF receptor regulation in breast cancer cells. Breast Cancer Res Treat 31: 227-236, 1994

    Google Scholar 

  47. Koenders PG, Beex LVAM, Geurts-Moespot A, Heuvel JJTM, Kienhuis CBM, Benraad TJ: Epidermal growth factor receptor-negative tumors are predominantly confined to the subgroup of estradiol receptor-positive human primary breast cancers. Cancer Res 51: 4544-4548, 1991

    Google Scholar 

  48. Ethier SP: Growth factor synthesis and human breast cancer progression. J Natl Cancer Inst 87: 964-973, 1995

    Google Scholar 

  49. Murphy LJ, Murphy LC, Freisen HG: Oestrogen induction of N-myc and c-myc proto-oncogene expression in the rat uterus. Endocrinology 120: 1882-1888, 1987

    Google Scholar 

  50. van der Burg B, van Selm-Mitenburg AJP, de Laat SW, van Zoelen EJJ: Direct effects of estrogen on c-fos and c-myc proto-oncogene expression and cellular proliferation in human breast cancer cells. Mol Cell Endocrinol 64: 223-228, 1989

    Google Scholar 

  51. Schuchard M, Landers JP, Sandhu NP, Spelsberg TC: Steroid hormone regulation of nuclear proto-oncogenes. Endocr Rev 14: 659-669, 1993

    Google Scholar 

  52. Dubik D, Dembinski TC, Shiu RP: Stimulation of c-myc expression associated with estrogen-induced proliferation of human breast cancer cells. Cancer Res 47: 6517-6521, 1987

    Google Scholar 

Download references

Authors
  1. Gabriela Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Graciela Cricco
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carlos Davio
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nora Engel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Claudia Cocca
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Elena Rivera
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rosa Bergoc
    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

Martin, G., Cricco, G., Davio, C. et al. Epidermal growth factor in NMU-induced mammary tumors in rats. Breast Cancer Res Treat 48, 175–185 (1998). https://doi.org/10.1023/A:1005994907827

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005994907827

Search

Navigation