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Journal List > Environ Health Perspect > v.105(Suppl 3); Apr 1997

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Environ Health Perspect. 1997 April; 105(Suppl 3): 619–624.
PMCID: PMC1469887
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Research Article
Neoplastic transformation of cultured mammalian cells by estrogens and estrogenlike chemicals.
T Tsutsui and J C Barrett
Department of Pharmacology, School of Dentistry at Tokyo, The Nippon Dental University, Japan.
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Abstract
Estrogens are clearly carcinogenic in humans and rodents but the mechanisms by which these hormones induce cancer are only partially understood. Stimulation of cell proliferation and gene expression by binding to the estrogen receptor is one important mechanism in hormonal carcinogenesis; however, estrogenicity is not sufficient to explain the carcinogenic activity of all estrogens because some estrogens are not carcinogenic. Estrogens are nonmutagenic in many assays but exhibit specific types of genotoxic activity under certain conditions. We have studied extensively the mechanisms by which estrogens induce neoplastic transformation in a model in vitro system and our findings are summarized in this review. 17beta-Estradiol (E2) and diethylstilbestrol (DES) and their metabolites induce morphological and neoplastic transformation of Syrian hamster embryo (SHE) cells that express no measurable levels of estrogen receptor. Treatment of the cells with E2 or DES fails to induce DNA damage, chromosome aberrations and gene mutations in SHE cells but results in numerical chromosome aberrations (aneuploidy) that could arise from microtubule disruption or disfunction of mitotic apparatus. Estrogen-induced genotoxicity is detected in cells following treatment with estrogen metabolites or following exogenous metabolic activation of estrogens. The estrogens induce DNA adduct formation that is detected by 32P-postlabeling. Both aneuploidy induction and DNA damage caused by DNA adduct formation correlate with the estrogen-induced cell transformation and may be important in hormonal carcinogenesis. We propose that multiple effects of estrogens acting together cause genetic alterations leading to cell transformation.
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Selected References
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  • Brinton LA, Hoover R, Fraumeni JF., Jr Menopausal oestrogens and breast cancer risk: an expanded case-control study. Br J Cancer. 1986 Nov;54(5):825–832. [PubMed]
  • KIRKMAN H. Estrogen-induced tumors of the kidney. III. Growth characteristics in the Syrian hamster. Natl Cancer Inst Monogr. 1959 Dec;1:1–57. [PubMed]
  • Herbst AL, Ulfelder H, Poskanzer DC. Adenocarcinoma of the vagina. Association of maternal stilbestrol therapy with tumor appearance in young women. N Engl J Med. 1971 Apr 15;284(15):878–881. [PubMed]
  • Barrett JC, Hesterberg TW, Oshimura M, Tsutsui T. Role of chemically induced mutagenic events in neoplastic transformation of Syrian hamster embryo cells. Carcinog Compr Surv. 1985;9:123–137. [PubMed]
  • Grubbs CJ, Peckham JC, McDonough KD. Effect of ovarian hormones on the induction of 1-methyl-1-nitrosourea-induced mammary cancer. Carcinogenesis. 1983;4(4):495–497. [PubMed]
  • Noronha RF, Goodall CM. The effects of estrogen on single dose dimethylnitrosamine carcinogenesis in male inbred Crl/CDF rats. Carcinogenesis. 1984 Aug;5(8):1003–1007. [PubMed]
  • Yager JD, Jr, Yager R. Oral contraceptive steroids as promoters of hepatocarcinogenesis in female Sprague-Dawley rats. Cancer Res. 1980 Oct;40(10):3680–3685. [PubMed]
  • Yager JD, Campbell HA, Longnecker DS, Roebuck BD, Benoit MC. Enhancement of hepatocarcinogenesis in female rats by ethinyl estradiol and mestranol but not estradiol. Cancer Res. 1984 Sep;44(9):3862–3869. [PubMed]
  • Sheehan DM, Frederick CB, Branham WS, Heath JE. Evidence for estradiol promotion of neoplastic lesions in the rat vagina after initiation with N-methyl-N-nitrosourea. Carcinogenesis. 1982;3(8):957–959. [PubMed]
  • Moolgavkar SH, Knudson AG., Jr Mutation and cancer: a model for human carcinogenesis. J Natl Cancer Inst. 1981 Jun;66(6):1037–1052. [PubMed]
  • Satyaswaroop PG, Zaino RJ, Mortel R. Human endometrial adenocarcinoma transplanted into nude mice: growth regulation by estradiol. Science. 1983 Jan 7;219(4580):58–60. [PubMed]
  • Barrett JC, Huff J. Cellular and molecular mechanisms of chemically induced renal carcinogenesis. Ren Fail. 1991;13(4):211–225. [PubMed]
  • Newbold RR, Bullock BC, McLachlan JA. Uterine adenocarcinoma in mice following developmental treatment with estrogens: a model for hormonal carcinogenesis. Cancer Res. 1990 Dec 1;50(23):7677–7681. [PubMed]
  • Barrett JC, Wong A, McLachlan JA. Diethylstilbestrol induces neoplastic transformation without measurable gene mutation at two loci. Science. 1981 Jun 19;212(4501):1402–1404. [PubMed]
  • Tsutsui T, Maizumi H, McLachlan JA, Barrett JC. Aneuploidy induction and cell transformation by diethylstilbestrol: a possible chromosomal mechanism in carcinogenesis. Cancer Res. 1983 Aug;43(8):3814–3821. [PubMed]
  • Tsutsui T, Degen GH, Schiffmann D, Wong A, Maizumi H, McLachlan JA, Barrett JC. Dependence on exogenous metabolic activation for induction of unscheduled DNA synthesis in Syrian hamster embryo cells by diethylstilbestrol and related compounds. Cancer Res. 1984 Jan;44(1):184–189. [PubMed]
  • Barrett JC, Oshimura M, Tanaka N, Tsutsui T. Role of aneuploidy in early and late stages of neoplastic progression of Syrian hamster embryo cells in culture. Basic Life Sci. 1985;36:523–538. [PubMed]
  • Tsutsui T, Suzuki N, Fukuda S, Sato M, Maizumi H, McLachlan JA, Barrett JC. 17beta-Estradiol-induced cell transformation and aneuploidy of Syrian hamster embryo cells in culture. Carcinogenesis. 1987 Nov;8(11):1715–1719. [PubMed]
  • McLachlan JA, Wong A, Degen GH, Barrett JC. Morphologic and neoplastic transformation of Syrian hamster embryo fibroblasts by diethylstilbestrol and its analogs. Cancer Res. 1982 Aug;42(8):3040–3045. [PubMed]
  • Ernst H, Riebe M, Mohr U. Undifferentiated sarcomas induced in Syrian hamsters by subcutaneous injection of diethylstilbestrol. Cancer Lett. 1986 May;31(2):181–186. [PubMed]
  • Korach KS, McLachlan JA. The role of the estrogen receptor in diethylstilbestrol toxicity. Arch Toxicol Suppl. 1985;8:33–42. [PubMed]
  • Degen GH, Wong A, Eling TE, Barrett JC, McLachlan JA. Involvement of prostaglandin synthetase in the peroxidative metabolism of diethylstilbestrol in Syrian hamster embryo fibroblast cell cultures. Cancer Res. 1983 Mar;43(3):992–996. [PubMed]
  • Hayashi N, Hasegawa K, Komine A, Tanaka Y, McLachian JA, Barrett JC, Tsutsui T. Estrogen-induced cell transformation and DNA adduct formation in cultured Syrian hamster embryo cells. Mol Carcinog. 1996 Jul;16(3):149–156. [PubMed]
  • Metzler M, McLachlan JA. Peroxidase-mediated oxidation, a possible pathway for metabolic activation of diethylstilbestrol. Biochem Biophys Res Commun. 1978 Dec 14;85(3):874–884. [PubMed]
  • Tsutsui T, Suzuki N, Maizumi H, McLachlan JA, Barrett JC. Alteration in diethylstilbestrol-induced mutagenicity and cell transformation by exogenous metabolic activation. Carcinogenesis. 1986 Sep;7(9):1415–1418. [PubMed]
  • Fitzgerald DJ, Piccoli C, Yamasaki H. Detection of non-genotoxic carcinogens in the BALB/c 3T3 cell transformation/mutation assay system. Mutagenesis. 1989 Jul;4(4):286–291. [PubMed]
  • Rinehart CA, Xu LH, Van Le L, Kaufman DG. Diethylstilbestrol-induced immortalization of human endometrial cells: alterations in p53 and estrogen receptor. Mol Carcinog. 1996 Feb;15(2):115–123. [PubMed]
  • Fishman J, Hellman L, Zumoff B, Cassouto J. Pathway and stereochemistry of the formation of estriols in man. Biochemistry. 1966 Jun;5(6):1789–1794. [PubMed]
  • Fishman J. Aromatic hydroxylation of estrogens. Annu Rev Physiol. 1983;45:61–72. [PubMed]
  • Li SA, Klicka JK, Li JJ. Estrogen 2- and 4-hydroxylase activity, catechol estrogen formation, and implications for estrogen carcinogenesis in the hamster kidney. Cancer Res. 1985 Jan;45(1):181–185. [PubMed]
  • Liehr JG. Genotoxic effects of estrogens. Mutat Res. 1990 May;238(3):269–276. [PubMed]
  • Telang NT, Suto A, Wong GY, Osborne MP, Bradlow HL. Induction by estrogen metabolite 16 alpha-hydroxyestrone of genotoxic damage and aberrant proliferation in mouse mammary epithelial cells. J Natl Cancer Inst. 1992 Apr 15;84(8):634–638. [PubMed]
  • Yu SC, Fishman J. Interaction of histones with estrogens. Covalent adduct formation with 16 alpha-hydroxyestrone. Biochemistry. 1985 Dec 31;24(27):8017–8021. [PubMed]
  • Swaneck GE, Fishman J. Covalent binding of the endogenous estrogen 16 alpha-hydroxyestrone to estradiol receptor in human breast cancer cells: characterization and intranuclear localization. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7831–7835. [PubMed]
  • Schneider J, Kinne D, Fracchia A, Pierce V, Anderson KE, Bradlow HL, Fishman J. Abnormal oxidative metabolism of estradiol in women with breast cancer. Proc Natl Acad Sci U S A. 1982 May;79(9):3047–3051. [PubMed]
  • Telang NT, Bradlow HL, Kurihara H, Osborne MP. In vitro biotransformation of estradiol by explant cultures of murine mammary tissues. Breast Cancer Res Treat. 1989 Mar;13(2):173–181. [PubMed]
  • Liehr JG, Purdy RH, Baran JS, Nutting EF, Colton F, Randerath E, Randerath K. Correlation of aromatic hydroxylation of 11 beta-substituted estrogens with morphological transformation in vitro but not with in vivo tumor induction by these hormones. Cancer Res. 1987 May 15;47(10):2583–2588. [PubMed]
  • Tsutsui T, Taguchi S, Tanaka Y, Barrett JC. 17beta-estradiol, diethylstilbestrol, tamoxifen, toremifene and ICI 164,384 induce morphological transformation and aneuploidy in cultured Syrian hamster embryo cells. Int J Cancer. 1997 Jan 17;70(2):188–193. [PubMed]
  • Wakeling AE, Bowler J. Biology and mode of action of pure antioestrogens. J Steroid Biochem. 1988;30(1-6):141–147. [PubMed]
  • Ozawa N, Oshimura M, McLachlan JA, Barrett JC. Nonrandom karyotypic changes in immortal and tumorigenic Syrian hamster cells induced by diethylstilbestrol. Cancer Genet Cytogenet. 1989 Apr;38(2):271–282. [PubMed]
  • Tucker RW, Barrett JC. Deceased numbers of spindle and cytoplasmic microtubules in hamster embryo cells treated with a carcinogen, diethylstilbestrol. Cancer Res. 1986 Apr;46(4 Pt 2):2088–2095. [PubMed]
  • Sakakibara Y, Saito I, Ichinoseki K, Oda T, Kaneko M, Saitô H, Kodama M, Sato Y. Effects of diethylstilbestrol and its methyl ethers on aneuploidy induction and microtubule distribution in Chinese hamster V79 cells. Mutat Res. 1991 Aug;263(4):269–276. [PubMed]
  • Aizu-Yokota E, Ichinoseki K, Sato Y. Microtubule disruption induced by estradiol in estrogen receptor-positive and -negative human breast cancer cell lines. Carcinogenesis. 1994 Sep;15(9):1875–1879. [PubMed]
  • Sato Y, Sakakibara Y, Oda T, Aizu-Yokota E, Ichinoseki K. Effect of estradiol and ethynylestradiol on microtubule distribution in Chinese hamster V79 cells. Chem Pharm Bull (Tokyo). 1992 Jan;40(1):182–184. [PubMed]
  • Aizu-Yokota E, Susaki A, Sato Y. Natural estrogens induce modulation of microtubules in Chinese hamster V79 cells in culture. Cancer Res. 1995 May 1;55(9):1863–1868. [PubMed]
  • Sato Y, Murai T, Tsumuraya M, Saitô H, Kodama M. Disruptive effect of diethylstilbestrol on microtubules. Gann. 1984 Dec;75(12):1046–1048. [PubMed]
  • Epe B, Hegler J, Metzler M. Site-specific covalent binding of stilbene-type and steroidal estrogens to tubulin following metabolic activation in vitro. Carcinogenesis. 1987 Sep;8(9):1271–1275. [PubMed]
  • Bradley MO, Bhuyan B, Francis MC, Langenbach R, Peterson A, Huberman E. Mutagenesis by chemical agents in V79 chinese hamster cells: a review and analysis of the literature. A report of the Gene-Tox Program. Mutat Res. 1981 Sep;87(2):81–142. [PubMed]
  • Hsie AW, Puck TT. Morphological transformation of Chinese hamster cells by dibutyryl adenosine cyclic 3':5'-monophosphate and testosterone. Proc Natl Acad Sci U S A. 1971 Feb;68(2):358–361. [PubMed]
  • Browne CL, Lockwood AH, Su JL, Beavo JA, Steiner AL. Immunofluorescent localization of cyclic nucleotide-dependent protein kinases on the mitotic apparatus of cultured cells. J Cell Biol. 1980 Nov;87(2 Pt 1):336–345. [PubMed]
  • Edwards KJ, Laughton CA, Neidle S. A molecular modeling study of the interactions between the antiestrogen drug tamoxifen and several derivatives, and the calcium-binding protein calmodulin. J Med Chem. 1992 Jul 24;35(15):2753–2761. [PubMed]
  • Sargent LM, Dragan YP, Bahnub N, Wiley JE, Sattler CA, Schroeder P, Sattler GL, Jordan VC, Pitot HC. Tamoxifen induces hepatic aneuploidy and mitotic spindle disruption after a single in vivo administration to female Sprague-Dawley rats. Cancer Res. 1994 Jul 1;54(13):3357–3360. [PubMed]
  • Geiser JR, Sundberg HA, Chang BH, Muller EG, Davis TN. The essential mitotic target of calmodulin is the 110-kilodalton component of the spindle pole body in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Dec;13(12):7913–7924. [PubMed]
  • Wheeler WJ, Hsu TC, Tousson A, Brinkley BR. Mitotic inhibition and chromosome displacement induced by estradiol in Chinese hamster cells. Cell Motil Cytoskeleton. 1987;7(3):235–247. [PubMed]
  • Schmuck G, Lieb G, Wild D, Schiffmann D, Henschler D. Characterization of an in vitro micronucleus assay with Syrian hamster embryo fibroblasts. Mutat Res. 1988 Dec;203(6):397–404. [PubMed]
  • Schiffmann D, De Boni U. Dislocation of chromatin elements in prophase induced by diethylstilbestrol: a novel mechanism by which micronuclei can arise. Mutat Res. 1991 Jan;246(1):113–122. [PubMed]
  • Schnitzler R, Foth J, Degen GH, Metzler M. Induction of micronuclei by stilbene-type and steroidal estrogens in Syrian hamster embryo and ovine seminal vesicle cells in vitro. Mutat Res. 1994 Nov 1;311(1):84–93. [PubMed]
  • Bahari IB, Noor FM, Daud NM. Micronucleated erythrocytes as an assay to assess actions by physical and chemical genotoxic agents in Clarias gariepinus. Mutat Res. 1994 Aug;313(1):1–5. [PubMed]
  • Foth J, Degen GH. Prostaglandin H synthase dependent metabolism of diethylstilbestrol by ram seminal vesicle cell cultures. Arch Toxicol. 1991;65(4):344–347. [PubMed]
  • Martin CN, McDermid AC, Garner RC. Testing of known carcinogens and noncarcinogens for their ability to induce unscheduled DNA synthesis in HeLa cells. Cancer Res. 1978 Aug;38(8):2621–2627. [PubMed]
  • Rüdiger HW, Haenisch F, Metzler M, Oesch F, Glatt HR. Metabolites of diethylstilboestrol induce sister chromatid exchange in human cultured fibroblasts. Nature. 1979 Oct 4;281(5730):392–394. [PubMed]
  • Hill A, Wolff S. Increased induction of sister chromatid exchange by diethylstilbestrol in lymphocytes from pregnant and premenopausal women. Cancer Res. 1982 Mar;42(3):893–896. [PubMed]
  • Buenaventura SK, Jacobson-Kram D, Dearfield KL, Williams JR. Induction of sister chromatid exchange by diethylstilbestrol in metabolically competent hepatoma cell lines but not in fibroblasts. Cancer Res. 1984 Sep;44(9):3851–3855. [PubMed]
  • Ivett JL, Tice RR. Diethylstilbestrol-diphosphate induces chromosomal aberrations but not sister chromatid exchanges in murine bone marrow cells in vivo. Environ Mutagen. 1981;3(4):445–452. [PubMed]
  • Clive D, Johnson KO, Spector JF, Batson AG, Brown MM. Validation and characterization of the L5178Y/TK+/- mouse lymphoma mutagen assay system. Mutat Res. 1979 Jan;59(1):61–108. [PubMed]
  • Liehr JG, Avitts TA, Randerath E, Randerath K. Estrogen-induced endogenous DNA adduction: possible mechanism of hormonal cancer. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5301–5305. [PubMed]
  • Gladek A, Liehr JG. Mechanism of genotoxicity of diethylstilbestrol in vivo. J Biol Chem. 1989 Oct 5;264(28):16847–16852. [PubMed]
  • Epe B, Schiffmann D, Metzler M. Possible role of oxygen radicals in cell transformation by diethylstilbestrol and related compounds. Carcinogenesis. 1986 Aug;7(8):1329–1334. [PubMed]
  • Liehr JG, Ulubelen AA, Strobel HW. Cytochrome P-450-mediated redox cycling of estrogens. J Biol Chem. 1986 Dec 25;261(36):16865–16870. [PubMed]
  • Roy D, Liehr JG. Temporary decrease in renal quinone reductase activity induced by chronic administration of estradiol to male Syrian hamsters. Increased superoxide formation by redox cycling of estrogen. J Biol Chem. 1988 Mar 15;263(8):3646–3651. [PubMed]
  • Liehr JG, Roy D. Free radical generation by redox cycling of estrogens. Free Radic Biol Med. 1990;8(4):415–423. [PubMed]
  • Rosier JA, Van Peteghem CH. Peroxidative in vitro metabolism of diethylstilbestrol induces formation of 8-hydroxy-2'-deoxyguanosine. Carcinogenesis. 1989 Feb;10(2):405–406. [PubMed]
  • Roy D, Floyd RA, Liehr JG. Elevated 8-hydroxydeoxyguanosine levels in DNA of diethylstilbestrol-treated Syrian hamsters: covalent DNA damage by free radicals generated by redox cycling of diethylstilbestrol. Cancer Res. 1991 Aug 1;51(15):3882–3885. [PubMed]
  • Han X, Liehr JG. 8-Hydroxylation of guanine bases in kidney and liver DNA of hamsters treated with estradiol: role of free radicals in estrogen-induced carcinogenesis. Cancer Res. 1994 Nov 1;54(21):5515–5517. [PubMed]
  • Haaf H, Metzler M. Covalent binding of diethylstilbestrol to microsomal protein in vitro correlates with the organotropism of its carcinogenicity. Carcinogenesis. 1985 Apr;6(4):659–660. [PubMed]
  • Epe B, Harttig U, Stopper H, Metzler M. Covalent binding of reactive estrogen metabolites to microtubular protein as a possible mechanism of aneuploidy induction and neoplastic cell transformation. Environ Health Perspect. 1990 Aug;88:123–127. [PubMed]
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