Breast Cancer Research and Treatment

, Volume 17, Issue 3, pp 197–210 | Cite as

Multiple actions of synthetic ‘progestins’ on the growth of ZR-75-1 human breast cancer cells: Anin vitro model for the simultaneous assay of androgen, progestin, estrogen, and glucocorticoid agonistic and antagonistic activities of steroids

  • Richard Poulin
  • Denis Baker
  • Donald Poirier
  • Fernand Labrie


This study was designed to assess the multiple steroid receptor mediated activities of a series of synthetic ‘progestins’ on breast cancer cell growth, using the human ZR-75-1 cell line which possesses functional estrogen (ER), androgen (AR), and glucocorticoid (GR) receptors as well as progesterone (PgR) receptors. Four 17-hydroxyprogesterone derivatives (chlormadinone acetate, CMA; cyproterone acetate, CPA; medroxyprogesterone acetate, MPA; and megestrol acetate, MGA) and two 19-nortestosterone derivatives (norethindrone, NRE, and norgestrel, NRG) were thus investigated.

Based on the requirement of estrogens for PgR-mediated antiproliferative effects and the reversal of PgR-mediated action by insulin, it was found that although all ‘progestins’ could inhibit ZR-75-1 cell growth through the PgR at low concentrations, the relative contribution of this receptor in cell growth control is highly variable between compounds. The quantitative importance of PgR-mediated inhibition of cell proliferation was inversely related to the amplitude of the androgenic effects induced by the compounds, the AR-mediated effects increasing in the order CPA < MGA < CMA < NRE < NRG < MPA. The specificity of these androgenic effects is further supported by their reversal upon addition of the antiandrogen hydroxyflutamide. In addition, the 17-hydroxyprogesterone derivatives, but not the 19-nortestosterone derivatives, had glucocorticoid activities at high (micromolar) concentrations, as shown by reversal of growth inhibition by the antagonist RU486 in the presence of saturating concentrations of 5α-dihydrotestosterone. All ‘progestins’ tested, except MPA and NRE, also had some antiglucocorticoid activity, NRG being the most potent in this respect. Finally, NRE and NRG exerted a marked mitogenic effect in estrogen-free medium which was clearly mediated through the ER as shown by the competitive reversal of their action by the steroidal antiestrogen EM-139.

The present results show that growth measurements of the human breast cancer cells ZR-75-1 permit, with the appropriate steroid additions, the assay of progestin, androgen, estrogen, and glucocorticoid agonistic as vell as antagonistic activities of test compounds. The present study shows, somewhat surprisingly, that while the AR is almost completely responsible for the action of MPA at low concentrations, the majority of the action of NRE, NRG, and MGA is also exerted through AR, while the androgenic action of CPA plays a lower role in the growth inhibition induced by this compound. Such a model should be of great help in designing more specific steroid drugs and in better understanding the role of the different steroid classes which can be used to control the growth of hormone-sensitive cancer. The present data also indicate that ‘progestin’ is an inappropriate name for MPA, NRE, NRG, MGA, CMA, and CPA, which all possess other and sometimes more potent steroidal activites than those related to interaction with the progesterone receptor.

Key words

progestins steroid receptors breast cancer androgens estrogens glucocorticoids ZR-75-1 cells 



chlormadinone acetate [17α-acetoxy-6-chloropregna-4, 6-dien-3, 20-dione]


cyproterone acetate [17α-acetoxy-6-chloro-1α,2α-methylene-pregna-4, 6-dien-3, 20-dione]


dexamethasone [9-fluoro-11β, 17, 21-trihydroxy-16α-methyl-pregna-1, 4-dien-3, 20-dione]


5α-dihydrotestosterone [17β-hydroxy-5β-androstan-3-one]


estradiol [estra-1, 3, 5 (10)-trien-3, 17β-diol]

EM 139

[N-n-butyl-N-methyl-11-(16α-chloro-3, 17β-dihydroxyestra-1, 3, 5 (10)-triene-7α-yl) undecanamide]


megestrol acetate [17α-acetoxy-6-methylpregna-4, 6-dien-3, 20-dionel]


medroxyprogesterone acetate [17α-acetoxy-6-methylpregn-4-en-3, 20-dione]


norethindrone [17β-hydroxy-19-nor-17α-pregn-4-en-20-yn-3-one]


norgestrel [13β-ethyl-17β-hydroxy-18, 19-dinor-17α-pregn-4-en-20-yn-3-one]


hydroxyflutamide (SCH 16423) [α, α, α-trifluoro-2-methyl-4′-nitro-m-lactotoluidide]


methyltrienolone [17β-hydroxy-17α-methyl estra-4, 9, 11-trien-3-one]


promegestone [17α, 21-dimethyl-19-norpregna-4, 9-dien-3, 20-dione]


[17β-hydroxy-11β-(4-dimethylaminophenyl)-17α-(1-propynyl)-estra-4, 9-dien-3-one]

triamcinolone acetonide

[9-fluoro-11β, 21-dihydroxy-16α, 17(1-methylethylidenebis 〈oxy〉) pregna-1, 4-dien-3, 20-dione]


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  1. 1.
    Haller DG, Glick JH: Progestational agents in advanced breast cancer: an overview. Semin Oncol 13 (suppl): 2–8, 1986PubMedGoogle Scholar
  2. 2.
    Furr BJA, Jordan VC: The pharmacology and clinical uses of tamoxifen. Pharmacol Ther 25: 127–205, 1984PubMedGoogle Scholar
  3. 3.
    Horwitz KB, Wei LL, Sedlacek SM, d'Arville CN: Progestin action and progesterone receptor structure in human breast cancer: a review: Rec Progr Horm Res 41: 249–316, 1985PubMedGoogle Scholar
  4. 4.
    Huggins C, Yang NC: Induction and extinction of mammary cancer. Science 137: 257–262, 1962PubMedGoogle Scholar
  5. 5.
    Jabara AG: Effects of progesterone of 9,10-dimethylbenz-(a)anthracene in Sprague-Dawley rats. Br J Cancer 27: 63–71, 1973PubMedGoogle Scholar
  6. 6.
    Asselin J, Kelly PA, Caron MG, Labrie F: Control of hormone receptor levels and growth of 7,12-dimethylbenz-(a)anthracene-induced mammary tumors by estrogens, progesterone, and prolactin. Endocrinology 101: 666–671, 1977PubMedGoogle Scholar
  7. 7.
    Leung BS, Potter AH, Qureshi S: Interactions of prolactin, estrogen, and progesterone in a human mammary carcinoma cell line, CAMA-1-I. Cell growth and thymidine uptake. J Steroid Biochem 15: 421–427, 1981PubMedGoogle Scholar
  8. 8.
    Hissom JR, Moore MR: Progestin effects on growth in the human breast cancer cell line T-47D possible therapeutic implications. Biochem Biophys Res Commun 145: 706–711, 1987PubMedGoogle Scholar
  9. 9.
    Vignon F, Bardon S, Chalbos D, Rochefort H: Antiestrogenic effect of R5020, a synthetic progestin human breast cancer cells in culture. J Clin Endocrinol Metab 56: 1124–1130, 1983PubMedGoogle Scholar
  10. 10.
    Horwitz KB, Freidenberg GR: Growth inhibition and increase of insulin receptor in antiestrogen-resistant T47Dco human breast cancer cells by ‘progestins’: implications for endocrine therapies. Cancer Res 45: 167–173, 1985PubMedGoogle Scholar
  11. 11.
    Engel LW, Young NA, Tralka TS, Lippman ME, O'Brien SJ, Joyce MJ: Establishment and characterization of three new continuous cell lines derived from human breast carcinomas. Cancer Res 38: 3352–3364, 1978PubMedGoogle Scholar
  12. 12.
    Poulin R, Dufour JM, Labrie F: Progestin inhibition of estrogen-dependent proliferation in ZR-75-1 human breast cancer cells: antagonism by insulin. Breast Cancer Res Treat 13: 265–276, 1989PubMedGoogle Scholar
  13. 13.
    Labrie F, Ferland L, Lagacé L, Drouin J, Asselin J, Azadian-Boulanger G, Raynaud JP: High inhibitory activity of R5020, a pure progestin, at the hypothalamic-adenohypophyseal level on gonadotropin secretion. Fertil Steril 28: 1104–1112, 1977PubMedGoogle Scholar
  14. 14.
    Raynaud JP, Ojasoo T, Labrie F: Steroid hormones agonists and antagonists. In: Lewis GP, Grisburg M (eds) Mechanisms of Steroid Action. MacMillan Press, London 1981, pp 145–158Google Scholar
  15. 15.
    Poyet P, Labrie F: Comparison of the antiandrogenic/androgenic activities of flutamide, cyproterone acetate, and megestrol acetate. Mol Cell Endocrinol 42: 283–288, 1985PubMedGoogle Scholar
  16. 16.
    Labrie C, Cusan L, Plante M, Lapointe S, Labrie F: Analysis of the androgenic activity of synthetic ‘progestins’ currently used in the treatment of prostate cancer. J Steroid Biochem 28: 379–384, 1987PubMedGoogle Scholar
  17. 17.
    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, 1989PubMedGoogle Scholar
  18. 18.
    Luthy IA, Bégin DJ, Labrie F: Androgenic activity of synthetic progestins and spironolactone in androgen-sensitive mouse mammary carcinoma (Shionogi) cells in culture. J Steroid Biochem 31: 845–852, 1988PubMedGoogle Scholar
  19. 19.
    Labrie C, Simard J, Zhao HF, Pelletier G, Labrie F: Synthetic progestins stimulate prostatic binding protein messenger RNAs in the rat ventral prostate. Mol Cell Endocrinol 68: 169–179, 1990PubMedGoogle Scholar
  20. 20.
    Pike MC, Chilvers C: Oral contraceptives and breast cancer: the current controversy. J Royal Soc Health 105: 5–10, 1985Google Scholar
  21. 21.
    Poulin R, Labrie F: Stimulation of cell proliferation and estrogenic response by adrenal C195-steroids in the ZR-75-1 human breast cancer cell line. Cancer Res 46: 4933–4937, 1986PubMedGoogle Scholar
  22. 22.
    Dickson RB, Lippman ME: Estrogenic stimulation of growth and polypeptide growth factor secretion in human breast carcinoma. Endocr Rev 8: 29–43, 1987PubMedGoogle Scholar
  23. 23.
    Poulin R, Baker D, Labrie F: Androgens inhibit basal and estrogen-induced cell proliferation in the ZR-75-1 human breast cancer cell line. Breast Cancer Res Treat 12: 213–225, 1988PubMedGoogle Scholar
  24. 24.
    Osborne CK, Monaco ME, Kahn CR, Huff K, Bronzert D, Lippman ME: Direct inhibition of growth and antagonism by glucocorticoids in human breast cancer cells in culture. Cancer Res 39: 2422–2428, 1979PubMedGoogle Scholar
  25. 25.
    Sutherland RL, Hall RE, Pang GYN, Musgrove EA, Clarke CL: Effect of medroxyprogesterone acetate on proliferation and cell cycle kinetics of human mammary carcinoma cells. Cancer Res 48: 5084–5091, 1988PubMedGoogle Scholar
  26. 26.
    Neri R, Peets E, Watnick A: Antiandrogenicity of flutamide and its metabolite SCH16423. Biochem Soc Trans 7: 565–569, 1979PubMedGoogle Scholar
  27. 27.
    Simard J, Luthy I, Guay J, Bélanger A, Labrie F: Characteristics of the interaction of the antiandrogen Flutamide with the androgen receptor in various target tissues. Mol Cell Endocrinol 44: 261–270, 1986PubMedGoogle Scholar
  28. 28.
    Philibert D: RU38486: an original multifaceted antihormonein vivo. In: Agarwal MK (ed) Adrenal Steroid Antagonism. Walter de Gruyter, Berlin, 1984, pp 77–101Google Scholar
  29. 29.
    Danhaive PA, Rousseau GG: Binding of glucocorticoid antagonists to androgen and glucocorticoid hormone receptors in rat skeletal muscle. J Steroid Biochem 24: 481–487, 1986PubMedGoogle Scholar
  30. 30.
    Lévesque C, Mérand Y, Dufour JM, Labrie C, Labrie F: Synthesis and biological activity of new halosteroidal antiestrogens. J Med Chem, 1991, in press.Google Scholar
  31. 31.
    Hubert JF, Vincent A, Labrie F: Estrogenic activity of phenol red in rat anterior pituitary cells in culture. Biochem Biophys Res Commun 141: 885–891, 1986PubMedGoogle Scholar
  32. 32.
    Taylor CM, Blanchard B, Zava DT: A simple method to determine whole cell uptake of radiolabeled estrogens and progesterone and their subcellular localization in breast cancer cell lines in monolayer cultures. J Steroid Biochem 20: 1083–1088, 1984PubMedGoogle Scholar
  33. 33.
    Rodbard D: Apparent positive cooperative effect in cyclic AMP and corticosterone production by isolated adrenal cells in response to ACTH analogues. Endocrinology 94: 1427–1437, 1974PubMedGoogle Scholar
  34. 34.
    Cheng Y, Prusoff WH: Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50% inhibition (IC50) of an enzymatic reaction. Biochem Pharmacol 22: 3099–3108, 1973PubMedGoogle Scholar
  35. 35.
    Munson PJ, Rodbard D: An exact correction to the ‘Cheng-Prusoff’ correction. J Receptor Res 8: 533–546, 1988Google Scholar
  36. 36.
    Poulin R, Simard J, Labrie C, Petitclerc L, Dumont M, Lagacé L, Labrie F: Down-regulation of estrogen receptors by androgens in the ZR-75-1 human breast cancer cell line. Endocrinology 125: 392–399, 1989PubMedGoogle Scholar
  37. 37.
    MacIndoe JH, Etre LA: An antiestrogenic action of androgens in human breast cancer cells. J Clin Endocrinol Metab 53: 836–842, 1981PubMedGoogle Scholar
  38. 38.
    Vilchis F, Chavez B, Pérez AE, Garcia GA, Angeles A, Pérez-Palacios G: Evidence that a non-aromatizable metabolite of norethisterone induces estrogen-dependent pituitary progesterone receptors. J Steroid Biochem 24: 525–531, 1986PubMedGoogle Scholar
  39. 39.
    Larrea F, Vilchis F, Chavez B, Pérez-Palacios G: The metabolism of 19-nor contraceptive ‘progestins’ modulates their biological activity at the neuroendocrine level. J Steroid Biochem 27: 657–663, 1987PubMedGoogle Scholar
  40. 40.
    Braselton WE, Lin TJ, Ellegood JD, Mills TM, Mahesh VB: Accumulation of norethindrone and individual metabolites in human plasma during short-term and long-term administration of a contraceptive dosage. Am J Obstet Gynecol 133: 154–169, 1979PubMedGoogle Scholar
  41. 41.
    Sahlberg BL, Landgren BM, Axelson M: Metabolic profiles of endogenous and ethynyl steroids in plasma and urine from women during administration of oral contraceptives. J Steroid Biochem 26: 609–617, 1987PubMedGoogle Scholar
  42. 42.
    Sahlberg BL: The characterization of sulfated metabolites of norethindrone in human milk after oral administration of contraceptive steroids. J Steroid Biochem 26: 481–485, 1987PubMedGoogle Scholar
  43. 43.
    Perel E, Daniilescu D, Kharlip L, Blackstein ME, Killinger DW: The relationship between growth and androstenedione metabolism in four cell lines of human breast carcinoma cells in culture. Mol Cell Endocrinol 41: 197–203, 1985PubMedGoogle Scholar
  44. 44.
    Gerhards E, Hecker W, Hitze H, Nieuweboer B, Bellman O: Zum Stoffwechsel von Norethisteron (17α-Äthinyl-4-östren-17β-ol-3-on) und diesowie D-Norgestrel (18-methyl-17α-Äthinyl-4-östren-17β-ol-3-on) beim Menschen. Acta Endocrinol (Copenh) 68: 219–248, 1971Google Scholar
  45. 45.
    Bullock LP, Bardin CW, Sherman MR: Androgenic, antiandrogenic, and synandrogenic actions of ‘progestins’: role of steric and allosteric interactions with androgen receptors. Endocrinology 103: 1768–1782, 1978PubMedGoogle Scholar
  46. 46.
    Sandberg AA, Kirdani RY: Metabolism of natural and synthetic steroids used in cancer treatment. Pharmac Ther 36: 263–307, 1988Google Scholar
  47. 47.
    Horwitz KB, Pike AW, Gonzalez-Allen C, Fennessey PV: Progesterone metabolism in T47Dco human breast cancer cells. II. Intracellular metabolic path of progesterone and synthetic progestins. J Steroid Biochem 25: 911–916, 1986PubMedGoogle Scholar
  48. 48.
    Raynaud JP, Bouton MM, Moguilewsky M, Ojasoo T, Philibert D, Beck G, Labrie F, Mormon JP: Steroid hormone receptors and pharmacology. J Steroid Biochem 12: 143–157, 1980PubMedGoogle Scholar
  49. 49.
    Blossey HC, Wander HE, Kobberling J, Nagel GA: Pharmacokinetic and pharmocodynamic basis for the treatment of metastatic breast cancer with high-dose medroxyprogesterone acetate. Cancer 54: 1208–1215, 1984PubMedGoogle Scholar
  50. 50.
    Clavel B, Pichon MF, Pallud C, Milgrom E: Estradiol and progesterone receptors content and response to norethisterone treatment in advanced breast cancer. Eur J Cancer Clin Oncol 18: 821–826, 1982PubMedGoogle Scholar
  51. 51.
    Alexieva-Figusch J, Blankenstein MA, Hop WCJ, Klijn JGM, Lamberts SWJ, de Jong FH, Docter R, Adlercreutz H, Van Gilse HA: Treatment of metastatic breast cancer patients with different dosages of megestrol acetate: dose relations, metabolism and endocrine effects. Eur J Cancer Clin Oncol 20: 33–40, 1984PubMedGoogle Scholar
  52. 52.
    Ettinger DS, Allegra J, Bertino JR, et al: Megestrol acetate vs. tamoxifen in advanced breast cancer: correlation of hormone receptors and response. Semin Oncol 13: 9–14, 1986PubMedGoogle Scholar
  53. 53.
    Alexieva-Figusch J, Teulings FAG, Hop WCJ, Blonk-van der Wijst J, van Gilse HA: Steroid receptors in megestrol acetate therapy. Rec Results Cancer Res 91: 253–258, 1984Google Scholar
  54. 54.
    Lundgren S, Kvinnsland S, Utaaker E: Oral high-dose progestins as treatment for advanced breast cancer. Acta Oncol 28: 811–816, 1989PubMedGoogle Scholar
  55. 55.
    Wanger HE, Blossey Ch, Köbberling J, Nagel GA: Hochdosiertes Medroxyprogesteronacetat beim metastasierenden Mammakarzinom: Beziehung zwischen Krankheitsverlauf und Hormonprofilen. Klin Wochenschr 61: 553–560, 1983PubMedGoogle Scholar
  56. 56.
    Earl HM, Rubens RD, Knight RK, Hayward JL: Norethisterone acetate in the treatment of advanced breast cancer. Clin Oncol 10: 103–109, 1984PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1990

Authors and Affiliations

  • Richard Poulin
    • 1
  • Denis Baker
    • 1
  • Donald Poirier
    • 1
  • Fernand Labrie
    • 1
  1. 1.Medical Research Council Group in Molecular EndocrinologyCHUL Research Center and Laval UniversityQuebecCanada

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