Summary
This study describes the inhibitory effect of 5α-dihydrotestosterone (5α-DHT) and its precursors testosterone (T) and androst-4-ene-3,17-dione (Δ4-DIONE) on the growth of the estrogen-sensitive human breast cancer cell line ZR-75-1. In the absence of estrogens, cell proliferation measured after a 12-day incubation period was 50–60% inhibited by maximal concentrations of 5α-DHT, T, or Δ4-DIONE with half-maximal effects (IC50 values) observed at 0.10, 0.15 and 15 nM, respectively. This growth inhibition by androgens was due to an increase in generation time and a lowering of the saturation density of cell cultures. The antiestrogen LY156758 (300 nM) induced 25–30% inhibition of basal cell growth, its effect being additive to that of 5α-DHT. The mitogenic effect of 1 nM estradiol (E2) was completely inhibited by increasing concentrations of 5α-DHT with a potency (IC50 = 0.10 nM) similar to that measured when the androgen was used alone. E2 had a more repid effect on cell proliferation than 5α-DHT, the latter requiring at least 5 to 6 days to exert significant growth inhibition. As found in the absence of estrogens, maximal inhibition of cell proliferation in the presence of E2 was achieved by the combination of the antiestrogen and 5α-DHT. Supraphysiological concentrations of E2 (up to 1µM) were needed to completely reverse the growth inhibitory effect of a submaximal concentration of 5α-DHT (1 nM). The antiproliferative effect of androgens was competitively reversed by the antiandrogen hydroxyflutamide, thus indicating an androgen receptor-mediated mechanism. The present data suggest the potential benefits of an androgen-antiestrogen combination therapy in the endocrine management of breast cancer.
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References
Fels E: Treatment of breast cancer with testosterone propionate. A preliminary report. J Clin Endocrinol 4: 121–125, 1944
Segaloff A, Gordon D, Horwitt BN, Schlosser JV, Murison PJ: Hormonal therapy in cancer of the breast. 1. The effect of testosterone propionate therapy on clinical course and hormonal excretion. Cancer 4: 319–323, 1951
Cooperative Breast Cancer Group: Testosterone propionate therapy of breast cancer. J Amer Med Assoc 188: 1069–1072, 1964
Kennedy BJ: Fluoxymesterone therapy in treatment of advanced breast cancer. N Engl J Med 259: 673–675, 1958
Tormey DC, Lippman ME, Cassidy JG: Evaluation of Tamoxifen doses with and without fluoxymesterone in advanced breast cancer. Ann Intern Med 98: 139–143, 1983
Gordan GS, Halden A, Horn Y, Fuery JJ, Parsons RJ, Walter RM: Calusterone (7β,17α-dimethyltestosterone) as primary and secondary therapy of advanced breast cancer. Oncology 28: 138–146, 1973
Gordan GS: In: Kochakian CD (ed) Anabolic-androgenic steroids. Handbook of Experimental Pharmacology, vol 43. Springer-Verlag, New York, 1976, pp 499–513
Segaloff A: The use of androgens in the treatment of neoplastic disease. Pharmac Ther C2: 33–37, 1977
McGuire WL, Carbone PP, Sears ME, Escher GC: Estrogen receptors in human breast cancer: an overview. In: McGuire WL, Carbone PP, Vollmer EP (eds) Estrogen Receptors in Breast Cancer, Raven Press, New York, 1975, pp 1–7
Wittliff JL: Steroid binding proteins in normal and neoplastic mammary cells. In: Busch H (ed) Methods in Cancer Research, vol 11 Academic Press, New York, 1975, pp 298–304
Allegra JC, Lippman ME, Thompson EG, Simon R, Barlock A, Green L, Huff KK, Do HMT, Aitken SC: Distribution, frequency, and quantitative analysis of estrogen, progesterone, androgen, and glucocorticoid receptors in human breast cancer. Cancer Res 39: 1447–1454, 1979
Horwitz KB, Zava DT, Thilager AK, Jensen ET, McGuire WL: Steroid receptor analyses of nine human breast cancer cell lines. Cancer Res 38: 2434–2439, 1978
Lippman M, Bolan G, Huff K: The effects of androgens and antiandrogens on hormone-responsive human breast cancer in long-term tissue culture. Cancer Res 36: 4610–4618, 1976
Westley B, Rochefort H: A secreted glycoprotein induced by estrogen in human breast cancer cell lines. Cell 20: 353–362, 1980
Zava DT, McGuire WL: Human breast cancer: androgen action mediated by estrogen receptor. Science 199: 787–788, 1978
Zava DT, McGuire WL: Androgen action through estrogen receptor in a human breast cancer cell line. Endocrinology 103: 624–631, 1978
McIndoe JH, Etre LA: An antiestrogenic action of androgens in human breast cancer cells. J Clin Endocrinol Metab 53: 836–842, 1981
Shapiro E, Lippman ME: Onset of androgen action in MCF-7 human breast cancer cells is not accompanied by receptor depletion. J Steroid Biochem 22: 15–20, 1985
Chalbos D, Haagensen D, Parish T, Rochefort H: Identification and androgen regulation of two proteins released by T47D human breast cancer cells. Cancer Res 47: 2787–2792, 1987
Murphy LC, Tsuyuki D, Myal Y, Shiu RPC: Isolation and sequencing of a cDNA clone for a prolactin-inducible protein (PIP). J Biol Chem 262: 15236–15241, 1987
Simon WE, Palinke VG, Hölzel F:In vitro modulation of prolactin binding to human mammary carcinoma cells by steroid hormones and prolactin. J Clin Endocrinol Metab 60: 1243–1249, 1985
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, 1978
Neri R, Peets E, Watnick A: Antiandrogenicity of flutamide and its metabolite Sch16423. Biochem Soc Trans 7: 565–569, 1979
Simard J, Luthy I, Guay J, Bélanger A, Labrie F: Characteristics of interaction of the antiandrogen Flutamide with the androgen receptor in various target tissues. Mol Cell Endocrinol 44: 261–270, 1986
Clemens JA, Bennett DR, Black LJ, Jones CD: Effects of a new antiestrogen, keoxifene (LY156758), on growth of car-cinogen-induced mammary tumors and on LH and prolactin levels. Life Sci 32: 2869–2875, 1983
Simard J, Labrie F: Keoxifene shows pure antiestrogenic activity in pituitary gonadotrophs. Mol Cell Endocrinol 39: 141–144, 1985
Berthois Y, Katzenellenbogen JA, Katzenellenbogen BS: Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci USA 83: 2496–2500, 1986
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, 1986
Poulin R, Labrie F: Stimulation of cell proliferation and estrogenic response by adrenal C19-Δ5-steroids in the ZR-75-1 human breast cancer cell line. Cancer Res 46: 4933–4937, 1986
Taylor CM, Blanchard B, Zava DT: A simple method to determine whole cell uptake of radiolabeled oestrogens and progesterone and their subcellular localization in breast cancer cell lines in monolayer cultures. J Steroid Biochem 20: 1083–1088, 1984
Scatchard G: The attraction of proteins for small molecules and ions. Ann NY Acad Sci 51: 660–672, 1959
Rodbard D: Apparent positive cooperative effect in cyclic AMP and corticosterone production by related adrenal cells in response to ACTH analogs. Endocrinology 94: 1427–1437, 1974
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, 1973
Kramer CY: Extension of multiple-range test to group means with unique numbers of replications. Biometrics 12: 307–310, 1956
Rochefort H, Garcia M: The estrogenic and antiestrogenic activities of androgens in female target tissues. Pharmac Ther 23: 193–216, 1983
Garcia M, Rochefort H: Androgen on the oestrogen receptor: II. Correlation between nuclear translocation and uterine protein synthesis. Steroids 29: 111–126, 1977
Huggins C, Jensen EV, Cleveland AS: Chemical structure of steroids in relation to promotion of growth of the vagina and uterus of the hypophysectomized rat. J Exp Med 100: 225–240, 1954
Hilf R: Anabolic-androgenic steroids and experimental mammary tumors. In: Kochakian CD (ed) Anabolic-androgenic steroids. Handbook of Experimental Pharmacology, vol 43, pp 191–210, 1976
Garcia M, Rochefort H: Androgen effects mediated by estrogen receptor in 7,12-dimethylbenz(a)anthracene-in-duced rat mammary tumors. Cancer Res 38: 3922–3929, 1978
Lippman M, Bolan G, Huff K: The effects of estrogens and antiestrogens on hormone-responsive human breast cancer in long-term tissue culture. Cancer Res 36: 4595–4601, 1976
Reiner GCA, Katzenellenbogen BS, Bindal RD, Katzenellenbogen JA: Biological activity and receptor binding of a strongly interacting estrogen in human breast cancer cells. Cancer Res 44: 2302–2308, 1984
Raynaud JP, Ojasoo T: Receptor binding as a tool in the development of selective new bioactive steroids and nonsteroids.In: Harms AF (ed) Innovative Approaches in Drug Research. Elsevier, Amsterdam, 1986, pp 47–72
Abraham GE: Ovarian and adrenal contribution to peripheral androgens during the menstrual cycle. J Clin Endocrinol Metab 39: 340–346, 1974
Vermeulen A, Verdonck L: Factors affecting sex hormone levels in postmenopausal women. J Steroid Biochem 11: 899–904, 1979
Mistry P, Griffiths K, Maynard PV: Endogenous C19-steroids and estradiol levels in human primary breast tumor tissues and their correlation with androgen and estrogen receptors. J Steroid Biochem 24: 1117–1125, 1986
McIndoe JH, Woods GR, Lee FJ: The specific binding of androgens and the subsequent distribution of androgen receptor complexes within MCF-7 human breast cancer cells. Steroids 38: 439–452, 1981
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, 1985
Griffiths K, Jones D, Cameron EHD, Gleave EN, Forrest APM: Transformation of steroids by mammary cancer tissue.In: Dao TL (ed) Oestrogen Target Tissues and Neoplasia. University of Chicago Press, Chicago, 1972, pp 125–136
Perel E, Killinger DW: The metabolism of androstenedione and testosterone to C19-metabolites in normal breast, breast carcinoma, and benign prostatic hypertrophy tissue. J Steroid Biochem 19: 1135–1139, 1983
Ip M, Milholland RJ, Kim U, Rosen F: Androgen control of cytosol progesterone receptor levels in the MT-W9B transplantable mammary tumor in the rat. J Natl Cancer Inst 69: 673–691, 1982
Jellinek PH, Newcombe AM: Androgen receptor-mediated inhibition of estrogen-induced uterine peroxidase. J Steroid Biochem 19: 1713–1717, 1983
Li SA, Li JJ: Estrogen-induced progesterone receptor in the Syrian hamster kidney. I. Modulation by antiestrogens and androgens. Endocrinology 103: 2119–2128, 1978
Tam S-P, Archer TK, Deeley RG: Biphasic effects of estrogen on apolipoprotein synthesis in human hepatoma cells. Mechanism of antagonism by testosterone. Proc Natl Acad Sci USA 83: 3111–3115, 1986
Stover EP, Krishman AV, Feldman D: Estrogen downregulation of androgen receptors in cultured human mammary cancer cells (MCF-7). Endocrinology 120: 2597–2603, 1987
Taylor CM, Blanchard B, Zava DT: Estrogen-receptor mediated and cytotoxic effects of the antiestrogens tamoxifen and 4-hydroxytamoxifen. Cancer Res 44: 1409–1414, 1984
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, 1987
Strobl JS, Lippman ME: Prolonged retention of estradiol by human breast cancer cells in tissue culture. Cancer Res 39: 3319–3327, 1979
Huggins C, Briziarelli G, Sutton H Jr: Rapid induction of mammary carcinoma in the rat and the influence of hormone on the tumors. J Exp Med 109: 25–42, 1959
Heise E, Gorlich M: Growth and therapy of mammary tumors induced by 7,12-dimethylbenz(a)anthracene in rats. Br J Cancer 20: 539–545, 1966
Horwitz HB: The structure and function of progesterone receptors in breast cancer. J Steroid Biochem 27: 447–457, 1987
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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 Tr 12, 213–225 (1988). https://doi.org/10.1007/BF01805942
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DOI: https://doi.org/10.1007/BF01805942