Skip to main content
SpringerLink
Log in

Establishment of Two Hormone-responsive Mouse Mammary Carcinoma Cell Lines Derived from a Metastatic Mammary Tumor

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

Abstract

We report the establishment of two mouse mammary cancer cell lines, MC7-2A and MC7-2B obtained from a mouse mammary carcinoma induced by medroxyprogesterone acetate (MPA) and maintained by syngeneic transplantation in BALB/c mice. They are epithelial (express cytokeratins) and express both estrogen receptors alpha (ERα) and progesterone receptors (PRs) isoforms A and B (western blots). In vitro, MPA inhibited 3H-thymidine uptake, starting from concentrations as low as 10−13 M in MC7-2A and 1010 M in MC7-2B; the antiprogestin RU 486 exerted a stimulatory effect at 10−14 M in both cell lines; 17-β-estradiol (E2) also exerted a stimulatory effect starting at 10−10 M in MC7-2A and at 10−13 M in MC7-2B. When transplanted in syngeneic mice, both cell lines originated adenocarcinomas that gave rise to lung metastases within 3 months. In in vivo studies, in MC7-2A, the antiprogestin inhibited completely tumor growth, E2 induced a slight although significant (p 9 0.05) stimulatory effect and MPA stimulated tumor growth while MC7-2B cells were unresponsive to all treatments. ER and PR were also expressed in tumors as assessed by immunohistochemistry. Two marker chromosomes were identified by FISH as translocations between chromosomes 4 and 7, and between chromosomes X and 2; the third marker chromosome remains unidentified. All these markers were also present in the parental tumor. A new marker, a centric fusion of chromosomes 2, was acquired in both cell lines. Considering that there are very few murine breast carcinoma responsive cell lines, these cells represent new tools in which the regulatory effect of hormones can be studied.

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. Lanari C, Luthy I, Lamb CA, Fabris V, Pagano E, Helguero LA, Sanjuan N, Merani S, Molinolo AA: Five novel hormone-responsive cell lines derived from murine mammary ductal carcinomas: in vivo and in vitro effects of estrogens and progestins. Cancer Res 61: 293–302, 2001

    Google Scholar 

  2. Bernhardt G, Beckenlehner K, Spruss T, Schlemmer R, Reile H, Schonenberger H: Establishment and characterization of new murine breast cancer cell lines. Arch Pharm (Weinheim) 335: 55–68, 2002

    Google Scholar 

  3. Lanari C, Molinolo AA, Pasqualini CD: Induction of mammary adenocarcinomas by medroxyprogesterone acetate in BALB/c female mice. Cancer Lett 33: 215–223, 1986

    Google Scholar 

  4. Molinolo AA, Lanari C, Charreau EH, Sanjuan N, Pasqualini CD: Mouse mammary tumors induced by medroxyprogesterone acetate: immunohistochemistry and hormonal receptors. J Natl Cancer Inst 79: 1341–1350, 1987

    Google Scholar 

  5. Kordon E, Lanari C, Meiss R, Charreau E, Pasqualini CD: Hormone dependence of a mouse mammary tumor line induced in vivo by medroxyprogesterone acetate. Breast Cancer Res Treat 17: 33–43, 1990

    Google Scholar 

  6. Dran G, Luthy IA, Molinolo AA, Charreau EH, Pasqualini CD, Lanari C: Effect of medroxyprogesterone acetate (MPA) and serum factors on cell proliferation in primary cultures of an MPA-induced mammary adenocarcinoma. Breast Cancer Res Treat 35: 173–186, 1995

    Google Scholar 

  7. Kordon E, Lanari C, Molinolo AA, Elizalde PV, Charreau EH, Pasqualini CD: Estrogen inhibition of MPA-induced mouse mammary tumor transplants. Int J Cancer 49: 900–905, 1991

    Google Scholar 

  8. Montecchia MF, Lamb C, Molinolo AA, Luthy IA, Pazos P, Charreau E, Vanzulli S, Lanari C: Progesterone receptor involvement in independent tumor growth in MPA-induced murine mammary adenocarcinomas. J Steroid Biochem Mol Biol 68: 11–21, 1999

    Google Scholar 

  9. Vanzulli S, Efeyan A, Benavides F, Helguero L, Peters G, Shen J, Conti CJ, Lanari C, Molinolo A: p21, p27 and p53 in estrogen and antiprogestin-induced tumor regression of experimental mouse mammary ductal carcinomas. Carcinogenesis 23: 749–757, 2002

    Google Scholar 

  10. Seabright M: A rapid banding technique for human chromosomes. Lancet 2: 971–972, 1971

    Google Scholar 

  11. Commitee on Standardized Genetic Nomenclature for mice: Standard karyotype of the mouse, Mus musculus. J Hered 63: 69–72, 1972

    Google Scholar 

  12. Lowry OH, Rosebrough NJ, Farr AL: Protein measurements with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  13. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    Google Scholar 

  14. Sizemore SR, Cole RD: A test for hormonal responsiveness in a mammary epithelial cell line, NMuMg. In vitro 18: 668–674, 1982

    Google Scholar 

  15. Institute of Laboratory Animal Resources, Commission on Life Sciences National Research Council. Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, DC, 1996

    Google Scholar 

  16. Helguero L, Viegas MH, Asaithamby A, Shyamala G, Lanari C, Molinolo A: Progesterone receptor expression in medroxyprogesterone acetate-induced murine mammary carcinomas and response to endocrine treatment. Breast Cancer Res Treat, 2003

  17. Masamura S, Santner SJ, Heitjan DF, Santen RJ: Estrogen deprivation causes estradiol hypersensitivity in human breast cancer cells. J Clin Endocrinol Metab 80: 2918–2925, 1995

    Google Scholar 

  18. Chun TY, Gregg D, Sarkar DK, Gorski J: Differential regulation by estrogens of growth and prolactin synthesis in pituitary cells suggests that only a small pool of estrogen receptors is required for growth. Proc Natl Acad Sci USA 95: 2325–2330, 1998

    Google Scholar 

  19. Endoh H, Sasaki H, Maruyama K, Takeyama K, Waga I, Shimizu T, Kato S, Kawashima H: Rapid activation of MAP kinase by estrogen in the bone cell line. Biochem Biophys Res Commun 235: 99–102, 1997

    Google Scholar 

  20. Bentel JM, Birrell SN, Pickering MA, Holds DJ, Horsfall DJ, Tilley WD: Androgen receptor agonist activity of the synthetic progestin, medroxyprogesterone acetate, in human breast cancer cells. Mol Cell Endocrinol 154: 11–20, 1999

    Google Scholar 

  21. Classen S, Possinger K, Pelka-Fleischer R, Wilmanns W: Effect of onapristone and medroxyprogesterone acetate on the proliferation and hormone receptor concentration of human breast cancer cells. J Steroid Biochem Mol Biol 45: 315–319, 1993

    Google Scholar 

  22. Sutherland RL, Hall RE, Pang GY, Musgrove EA, Clarke CL: Effect of medroxyprogesterone acetate on proliferation and cell cycle kinetics of human mammary carcinoma cells. Cancer Res 48: 5084–5091, 1988

    Google Scholar 

  23. Hissom JR, Bowden RT, Moore MR: Effects of progestins, estrogens, and antihormones on growth and lactate dehydrogenase in the human breast cancer cell line T47D. Endocrinology 125: 418–423, 1989

    Google Scholar 

  24. Hackenberg R, Luttchens S, Hofmann J, Kunzmann R, Holzel F, Schulz KD: Androgen sensitivity of the new human breast cancer cell line MFM-223. Cancer Res 51: 5722–5727, 1991

    Google Scholar 

  25. Hackenberg R, Hawighorst T, Filmer A, Nia AH, Schulz KD: Medroxyprogesterone acetate inhibits the proliferation of estrogen-and progesterone-receptor negative MFM-223 human mammary cancer cells via the androgen receptor. Breast Cancer Res Treat 25: 217–224, 1993

    Google Scholar 

  26. Lin VC, Eng AS, Hen NE, Ng EH, Chowdhury SH: Effect of progesterone on the invasive properties and tumor growth of progesterone receptor-transfected breast cancer cells MDAMB-231. Clin Cancer Res 7: 2880–2886, 2001

    Google Scholar 

  27. Levenson AS, Jordan VC: Transfection of human estrogen receptor (ER) cDNA into ER-negative mammalian cell lines. J Steroid Biochem Mol Biol 51: 229–239, 1994

    Google Scholar 

  28. Thomas M, Monet JD: Combined effects of RU486 and tamoxifen on the growth and cell cycle phases of the MCF-7 cell line. J Clin Endocrinol Metab 75: 865–870, 1992

    Google Scholar 

  29. Bardon S, Vignon F, Chalbos D, Rochefort H: RU486, a progestin and glucocorticoid antagonist, inhibits the growth of breast cancer cells via the progesterone receptor. J Clin Endocrinol Metabolism 60: 692–697, 1985

    Google Scholar 

  30. Klijn JG, Setyono-Han B, Foekens JA: Progesterone antagonists and progesterone receptor modulators in the treatment of breast cancer. Steroids 65: 825–830, 2000

    Google Scholar 

  31. Moreno-Cuevas JE, Sirbasku DA: Estrogen mitogenic action. I. Demonstration of estrogen-dependent MTW9/PL2 carcinogen-induced rat mammary tumor cell growth in serumsupplemented culture and technical implications. In vitro Cell Dev Biol Anim 36: 410–427, 2000

    Google Scholar 

  32. Soto AM, Sonnenschein C: Cell proliferation of estrogen-sensitive cells: the case for negative control. Endocr Rev 8: 44–52, 1987

    Google Scholar 

  33. Gazdar AF, Kurvari V, Virmani A, Gollahon L, Sakaguchi M, Westerfield M, Kodagoda D, Stasny V, Cunningham HT, Wistuba II, Tomlinson G, Tonk V, Ashfaq R, Leitch AM, Minna JD, Shay JW: Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer. Int J Cancer 78: 766–774, 1998

    Google Scholar 

  34. Truong K, Guilly MN, Gerbault-Seureau M, Malfoy B, Vielh P, Dutrillaux B: Evidence for in vitro selection during cell culturing of breast cancer: detection by flow and image cytometry. Cancer Genet Cytogenet 114: 154–155, 1999

    Google Scholar 

  35. Fabris V, Lamb C, Keck C, Aldaz CM, Merani S, Lanari C: Karyotypic evolution of four novel mouse mammary carcinoma cell lines: identification of marker chromosomes by FISH. Cancer Genet Cytogenet, 2003

  36. Dietrich WF, Radany EH, Smith JS, Bishop JM, Hanahan D, Lander ES: Genome-wide search for loss of heterozygosity in transgenic mouse tumors reveals candidate tumor suppressor genes on chromosomes 9 and 16. Proc Natl Acad Sci USA 91: 9451–9455, 1994

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Hormonal Carcinogenesis, Instituto de Biología y Medicina Experimental – Consejo Nacional de Investigaciones Científicas y Técnicas (IBYME-CONICET), Buenos Aires, Argentina

    Alejo Efeyan, Victoria Fabris, Claudia Lanari & Alfredo A. Molinolo

  2. Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

    Susana Merani

Authors
  1. Alejo Efeyan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Victoria Fabris
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Susana Merani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Claudia Lanari
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alfredo A. Molinolo
    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

Efeyan, A., Fabris, V., Merani, S. et al. Establishment of Two Hormone-responsive Mouse Mammary Carcinoma Cell Lines Derived from a Metastatic Mammary Tumor. Breast Cancer Res Treat 83, 233–244 (2004). https://doi.org/10.1023/B:BREA.0000014044.02728.ac

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:BREA.0000014044.02728.ac

Search

Navigation