Breast Cancer Research and Treatment

, Volume 35, Issue 2, pp 173–186 | Cite as

Effect of medroxyprogesterone acetate (MPA) and serum factors on cell proliferation in primary cultures of an MPA-induced mammary adenocarcinoma

  • Graciela Dran
  • Isabel A. Luthy
  • Alfredo A. Molinolo
  • Fernanda Montecchia
  • Eduardo H. Charreau
  • Christiane Dosne Pasqualini
  • Claudia Lanari
Report

Summary

The effect of progesterone (Pg), medroxyprogesterone acetate (MPA), estradiol (E2), dihydrotestosterone (DHT) and dexamethasone (DEXA) was studied on thein vitro growth rate of a progestin-dependent (PD), estrogen-sensitive mammary tumor line originated in an MPA-treated BALB/c mouse (C4-HD), and on its estrogen-resistant variant (C4-HDR). The specificity of hormone action was further investigated using the anti-hormones RU-486 and hydroxyflutamide (FLU). Cell growth was evaluated in epithelial and fibroblastenriched cultures using3H-thymidine and/or autoradiography and immunocytochemistry. The results indicate that cell growth is directly stimulated by MPA and Pg at concentrations ranging from 10−11 to 10−7 M. RU486 prevented MPA-induced stimulation in concentrations 10 to 100 fold lower than those of MPA. When used alone, it inhibited cell proliferation only in concentrations higher than 10−11 M. At nM concentrations, neither DEXA nor DHT stimulated3H-thyrnidine uptake except DEXA at 100 nM. MPA-induced stimulation was not reverted by micromolar concentrations of FLU. As for E2 (10−7–10−9 M) it prevented MPA stimulation only in cultures of estrogen-sensitive tumors. Progesterone receptors (PR) (475 ± 115 fmoles/105 cells, n = 5) and estrogen receptors (ER) (ND-115 fmoles/105 cells, n = 5) were detected only in epithelial-enriched cultures. Serum from 7 day-MPA-treated mice induced a significant increase of3H-thymidine uptake; an increase was also obtained with serum from untreated ovariectomized animals to which 1 nM-100 nM concentrations of MPA had been added. The stimulatory effect of the exogenous MPA was much lower than that of the serum obtained from MPA-treated animals.

It is concluded that MPA stimulates cell growth of primary cultures of MPA-induced PD tumors via PR. The results provide support for a direct effect of MPA which may be mediated or potentiated by serum factors.

Key words

cell proliferation mammary tumors medroxyprogesterone acetate primary cultures progestins serum factors 

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References

  1. 1.
    Goustin AS, Leof EB, Shipley GD, Moses HL: Growth factors and cancer. Cancer Res 46: 1015–1029, 1986Google Scholar
  2. 2.
    Clarke CL, Sutherland RL: Progestin regulation of cellular proliferation. Endocrine Reviews 11: 266–300, 1990Google Scholar
  3. 3.
    Martin L, Finn CA: Hormonal regulation of cell division in epithelial and connective tissues in the mouse uterus. J Endocrinol 41: 363–371, 1968Google Scholar
  4. 4.
    Topper YJ, Freeman CS: Multiple hormone interactions in the developmental biology of the mammary gland. Physiol Rev 60: 1049–1106, 1980Google Scholar
  5. 5.
    Haslam SZ: Progesterone effects on deoxyribonucleic acid synthesis in the normal mouse mammary glands. Endocrinology 122: 464–470, 1988Google Scholar
  6. 6.
    Li JJ, Mueller GC, Sekeley LI: Meeting report: current perspectives and future trends in hormonal carcinogenesis. Cancer Res 51: 3626–3629, 1991Google Scholar
  7. 7.
    Imagawa W, Tomooka Y, Hamamoto S, Nandi S: Stimulation of mammary epithelial cell growthin vitro: Interaction of epidermal growth factor and mammogenic hormones. Endocrinology 116: 1514–1524, 1985Google Scholar
  8. 8.
    Robustelli Della Cuna G, Petri P: Advanced breast cancer: Hormone-dependency and mechanisms of action of medroxyprogesterone acetate. In: Robustelli Della Cuna G, Tepmongkol P, Somboonchareon (eds) The Role of High Dose Medroxyprogesterone Acetate in the Management of Endocrine-related Tumors, Bangkok, pp 21-27, 1986Google Scholar
  9. 9.
    Bonte J: Hormone dependency and hormone-responsiveness of endometrial adenocarcinoma to estrogens, progestogens and antioestrogens. In: Campio L, Robustelli Della Cuna G, Taylor RW (eds) Role of Medroxyprogesterone Acetate in Endocrine-Related Tumors. Raven Press, NY, 1983, Vol II. pp 141–156Google Scholar
  10. 10.
    Lanari A, Molinas FC, Castro Ríos M, Paz RA: Eficaz tratamiento de diversas fibromatosis con progesterona. Medicina (Bs. Aires) 38: 123–132, 1978Google Scholar
  11. 11.
    Danguy A, Legros N, Devleeschouwer N, Heuson-Stiennon JA, Heuson JC: Effects of medroxyprogesterone acetate (MPA) on growth of DMBA-induced rat mammary tumors: histopathological and endocrine studies. In: Iacobelli S, Di Marco A (eds) Role of Medroxyprogesterone Acetate in Endocrine Related Tumors. Raven Press, NY, 1980, pp 21–28Google Scholar
  12. 12.
    Nagasawa H, Aoki M, Sakagami N, Ishida M: Medroxyprogesterone acetate enhances spontaneous mammary tumorigenesis and uterine adenomyosis in mice. Breast Cancer Res Treat 12: 59–66, 1988Google Scholar
  13. 13.
    Russo IH, Gimotty P, Dupuis M, Russo J: Effect of medroxyprogesterone acetate on the response of the rat mammary gland to carcinogenesis. Br J Cancer 59: 210–216, 1989Google Scholar
  14. 14.
    Kiss R, Paridaens RJ, Heuson JC, Danguy AJ: Effect of progesterone on cell proliferation in the MXT mouse hormonesensitive mammary neoplasms. JNCI 77: 173–178, 1986Google Scholar
  15. 15.
    Nagasawa H, Yanai R, Nakajima Y, Mori J: Effects of progesterone on normal and preneoplastic development in mice in relation to prolactin and estrogen. Europ J Cancer 16: 1069–1077, 1980Google Scholar
  16. 16.
    Manni A, Badger B, Wright C, Ahmed SR, Demers LM: Effects of progestins on growth of experimental breast cancer in culture: Interaction with estradiol and prolactin and involvement of the polyamine pathway. Cancer Res 47: 3066–3071, 1987Google Scholar
  17. 17.
    Jabara AG, Toyne PH, Harcourt AG: Effects of time and duration of progesterone administration on mammary tumors induced by 7, 12 dimethylbenzanthracene in Sprague Dawley rats. Brit J Cancer 27: 63–71, 1973Google Scholar
  18. 18.
    Richards JE, Edery M, Osborn RC, Larson LN, Nandi S: Effect of hormones and epidermal growth factor on the growth of the hormone-responsive 13762NF rat mammary tumor in collagen gel culture. J Natl Cancer Inst 76: 669–682, 1986Google Scholar
  19. 19.
    Pasteels JL, Heuson J-C, Heuson-Stiennon J, Legros N: Effects of insulin, prolactin, progesterone, and estradiol on DNA synthesis in organ culture of 7,12-dimethylbenz(a) anthracene-induced rat mammary tumors. Cancer Res 36: 2162–2170, 1976Google Scholar
  20. 20.
    Sluyser M, Van Nie R: Estrogen receptor content and hormone responsive growth of mouse mammary tumors. Cancer Res 34: 3253–3257, 1974Google Scholar
  21. 21.
    Lanari C. Molinolo AA, Pasqualini CD: Induction of mammary adenocarcinomas by medroxyprogesterone acetate in BALB/c mice. Cancer Letters 33: 215–223, 1986Google Scholar
  22. 22.
    Molinolo AA, Lanari C, Charreau EH, Sanjuan N, Pasqualini CD: Mouse mammary tumors induced by medroxyprogesterone acetate: Immunochemistry and hormonal receptors. J Natl Cancer Inst 79: 1341–1350, 1987Google Scholar
  23. 23.
    Lanari C, Kordon E, Molinolo A, Pasqualini CD, Charreau EH: Mammary adenocarcinomas induced by medroxyprogesterone acetate: Hormone dependence and EGF receptors of BALB/cin vivo sublines. Int J Cancer 43: 845–850, 1989Google Scholar
  24. 24.
    Kordon E, Lanari C, Meiss R, Elizalde P, Charreau EH, Pasqualini CD: Hormone dependence of anin vivo medroxyprogesterone acetate-dependent tumor line of murine mammary cancer. Breast Cancer Res Treat 17: 33–43, 1990Google Scholar
  25. 25.
    Kordon E, Lanari C, Molinolo AA, Elizalde P, Charreau EH, Pasqualini CD: Estrogen inhibition of MPA-induced mouse mammary tumor transplants. Int J Cancer 49: 900–905, 1991Google Scholar
  26. 26.
    Soto AM, Sonnenschein C: The role of estrogens in the proliferation of human breast tumor cells (MCF-7). J Steroid Biochem 23: 87–94, 1985Google Scholar
  27. 27.
    Pandis N, Heim S, Bardi G, Limon J, Mandahl N, Mitelman F: Improved technique for short-term culture and cytogenetic analysis of human breast cancer. Genes, Chromosomes & Cancer 5: 14–20, 1992Google Scholar
  28. 28.
    Hsu SM, Raine L, Fanger H: A comparative study of the peroxidase-antiperoxidase method and an avidin-biotin complex for studying polypeptide hormones with radioimmunoassay antibodies. Am J Clin Pathol 75: 734–738, 1981Google Scholar
  29. 29.
    Luthy I, Begin D, Labrie F: Mediation by the androgen receptor of the stimulatory and antiandrogenic actions of 17-beta estradiol on the growth of androgen-sensitive Shionogi mammary carcinoma cells in culture. Endocrinology 123: 1418–1424, 1988Google Scholar
  30. 30.
    Kordon E, Guerra F, Molinolo A, Elizalde P, Charreau E, Pasaqualini D, Montecchia MF, Pazos P, Dran G, Lanari C: Effect of sialoadenectomy on medroxyprogesterone acetate-induced mammary carcinogenesis in BALB/c mice. Correlation between histology and epidermal growth factor receptor contents. Int J Cancer 59: 196–203, 1994Google Scholar
  31. 31.
    Lippman ME, Dickson RB: Mechanisms of growth control in normal and malignant breast epithelium. Recent Prog Horm Res 45: 383–440, 1989Google Scholar
  32. 32.
    Markaverich BM, Clark JH: Two binding sites for estradiol in rat uterine nuclei: relationship to uterotropic response. Endocrinology 105: 1458–1462, 1979Google Scholar
  33. 33.
    Olea Serrano N, Devleeschower N, Leclercq G, Heuson JC: Assay for estrogen and progesterone receptors of breast cancer cell lines in monolayer culture. Eur J Cancer Clin Oncol 21: 965–973, 1985Google Scholar
  34. 34.
    Briggs MH, Briggs M: Glucocorticoid properties of progestogens. Selection and characterization of a breast cancer cell line resistant to the antiestrogen LY 117018. Steroids 22: 555–559, 1973Google Scholar
  35. 35.
    Bullock LP, Barthe TH, Mawzowicz I, North A, Bardin CW: The effect of progestin on submaxillary gland epidermal growth factor. Demonstration of androgenic, synandrogenic and anti-androgenic actions. Endocrinology 97: 189–195, 1975Google Scholar
  36. 36.
    Poulin R, Baker D, Poirier D, Labrie F: Androgen and glucocorticoid receptor-mediated inhibition of cell proliferation by medroxyprogesterone acetate in ZR-75-1 human breast cancer cells. Breast Cancer Res Treat 13: 161–172, 1989Google Scholar
  37. 37.
    Hackenberg R, Hawighorst T, Filmer A, Nia AH, Schulz KD: Medroxyprogesterone acetate inhibits the proliferation of estrogen-receptor and progesterone receptor negative MFM-223 human mammary cancer cells via the androgen receptor. Breast Cancer Res and Treat 25: 217–224, 1993Google Scholar
  38. 38.
    Baulieu EE: Contragestion and other clinical applications of RU486, an antiprogesterone at the receptor. Science 245: 1351–1357, 1989Google Scholar
  39. 39.
    Beck CA, Estes PA, Bona BJ, Murocacho CA, Nordeen SK, Edwards DP: The steroid antagonist RU486 exerts different effects on the glucocorticoid and progesterone receptors. Endocrinology 13: 728–740, 1993Google Scholar
  40. 40.
    Bardon S, Vignon F, Montcourrier P, Rochefort H: Steroid-receptor mediated cytotoxicity of an antiestrogen and an antiprogestin in breast cancer cells. Cancer Res 47: 1441–1448, 1987Google Scholar
  41. 41.
    van der Burg B: Sex steroids and growth factors in mammary cancer. Acta Endocrinologica (Copenh) 1991: 125–127, 1991Google Scholar
  42. 42.
    Cho H, Aronica M, Katzenellenbogen BS: Regulation of progesterone gene expression in MCF-7 cells: A comparison of the effects of cyclic adenosine 3′,5′-monophosphate, estradiol, insulin-like growth factor-I, and serum factors. Endocrinology 134: 658–664, 1994Google Scholar
  43. 43.
    Edwards DP, Weigel NL, Nordeen SK, Beck CA: Modulators of cellular protein phosphorylation after the trans-activation function of human progesterone receptor and the biological activity of progesterone antagonists. Breast Cancer Res Treat 27: 41–56, 1993Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Graciela Dran
    • 1
  • Isabel A. Luthy
    • 1
  • Alfredo A. Molinolo
    • 1
  • Fernanda Montecchia
    • 1
  • Eduardo H. Charreau
    • 1
  • Christiane Dosne Pasqualini
    • 1
  • Claudia Lanari
    • 1
  1. 1.División Medicina Experimental, Instituto de Investigaciones HematológicasAcademia Nacional de Medicina & Instituto de Biología y Medicina ExperimentalBuenos AiresArgentina

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