Phytoestrogens Modulate the Expression of 17α-Estradiol Metabolizing Enzymes in Cultured MCF-7 Cells

  • Jörg Wagner
  • Ling Jiang
  • Leane Lehmann
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 617)


The activation of 17β-estradiol (E2) to 2-hydroxyestradiol (2-HO-E2), the more genotoxic 4-hydroxyestradiol (4-HO-E2), and the oxidation to the respective quinones constitutes a risk factor in hormonal carcinogenesis. 2-HO-E2 is formed by cytochrome P450 CYP1A1, and 4-HO-E2 is formed by CYP1B1. Both are detoxified by catechol-O-methyltransferase (COMT), whereas their quinones are inactivated by NADPH-quinone-oxidoreductase (QR). Since the soy isoflavones genistein (GEN) and daidzein (DAI) are widely consumed due to their putative protective function in breast carcinogenesis, we examined the influence of E2, GEN, and DAI on CYP1A1/1B1, COMT, and QR expression in MCF-7 cells by reverse transcription/competitive PCR. CYP1A1 and COMT enzyme activity were determined using ethoxyresorufin and quercetin as substrates. Furthermore, estrogen receptor (ER)-regulated cell proliferation was determined by E-screen. E2, GEN, and DAI inhibited the expression of CYP1A1, COMT, and QR. The maximum effect (reduction by 40–80%, depending on the gene product and compound) was obtained at 100pM E2,1 μM GEN, and 10μM DAI, which also induced the most pronounced cell proliferation in the E-screen. In contrast, expression of CYP1B1 was only slightly affected. CYP1A1 and COMT mRNA levels correlated with enzyme activities. The ER antagonist ICI 182,780 reversed the E2- and isoflavone-mediated effects. Thus, GEN and DAI at estrogen-active concentrations stimulate the formation of the more E2 genotoxic metabolites and inhibit the detoxification of catechol and quinone estrogens in estrogen-responsive tumor cells.


Relative mRNA Level COMT Activity Catechol Estrogen Antioxidant Responsive Element Genotoxic Metabolite 
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  1. 1.
    Cavalieri E, Chakravarti D, Guttenplan J, et al. (2006) Catechol estrogen quinones as initiators of breast and other human cancers: implications for biomarkers of susceptibility and cancer prevention. Biochim Biophys Acta 1766: 63–78.PubMedGoogle Scholar
  2. 2.
    Guillemette C, Belanger A, Lepine J (2004) Metabolic inactivation of estrogens in breast tissue by UDP-glucuronosyltransferase enzymes: an overview. Breast Cancer Res 6: 246–254.PubMedCrossRefGoogle Scholar
  3. 3.
    Badawi AF, Cavalieri EL, Rogan EG (2001) Role of human cytochrome P450 1A1, 1A2, 1B1, and 3A4 in the 2-, 4-, and 16alpha-hydroxylation of 17beta-estradiol. Metabolism 50: 1001–1003.PubMedCrossRefGoogle Scholar
  4. 4.
    Zhu BT, Conney AH (1998) Functional role of estrogen metabolism in target cells: review and perspectives. Carcinogenesis 19: 1–27.PubMedCrossRefGoogle Scholar
  5. 5.
    Ball P, Knuppen R (1980) Catecholoestrogens (2-and 4-hydroxyoestrogens): chemistry, biogenesis, metabolism, occurrence and physiological significance. Acta Endocrinol Suppl (Copenh) 232: 1–127.Google Scholar
  6. 6.
    Albin N, Massaad L, Toussaint C, et al. (1993) Main drug-metabolizing enzyme systems in human breast tumors and peritumoral tissues. Cancer Res 53: 3541–3546.PubMedGoogle Scholar
  7. 7.
    Dawling S, Hachey DL, Roodi N, et al. (2004) In vitro model of mammary estrogen metabolism: structural and kinetic differences between catechol estrogens 2- and 4-hydroxyestradiol. Chem Res Toxicol 17: 1258–1264.PubMedCrossRefGoogle Scholar
  8. 8.
    Danson S, Ward TH, Butler J, et al. (2004) DT-diaphorase: a target for new anticancer drugs. Cancer Treat Rev 30: 437–449.PubMedCrossRefGoogle Scholar
  9. 9.
    Adlercreutz H, Heinonen SM, Penalvo-Garcia J (2004) Phytoestrogens, cancer and coronary heart disease. Biofactors 22: 229–236.PubMedCrossRefGoogle Scholar
  10. 10.
    Cornwell T, Cohick W, Raskin I (2004) Dietary phytoestrogens and health. Phytochemistry 65: 995–1016.PubMedCrossRefGoogle Scholar
  11. 11.
    Limer JL, Speirs V (2004) Phyto-oestrogens and breast cancer chemoprevention. Breast Cancer Res 6: 119–127.PubMedCrossRefGoogle Scholar
  12. 12.
    Rogan EG, Badawi AF, Devanesan PD, et al. (2003) Relative imbalances in estrogen metabolism and conjugation in breast tissue of women with carcinoma: potential biomarkers of susceptibility to cancer. Carcinogenesis 24: 697–702.PubMedCrossRefGoogle Scholar
  13. 13.
    Singh S, Chakravarti D, Edney JA, et al. (2005) Relative imbalances in the expression of estrogen-metabolizing enzymes in the breast tissue of women with breast carcinoma. Oncol Rep 14: 1091–1096.PubMedGoogle Scholar
  14. 14.
    Mitrunen K, Hirvonen A (2003) Molecular epidemiology of sporadic breast cancer. The role of polymorphic genes involved in oestrogen biosynthesis and metabolism. Mutat Res 544: 9–41.PubMedCrossRefGoogle Scholar
  15. 15.
    Zhu BT, Liehr JG (1996) Inhibition of catechol O-methyltransferase-catalyzed O-methylation of 2- and 4-hydroxyestradiol by quercetin. Possible role in estradiol-induced tumorigenesis. J Biol Chem 271: 1357–1363.PubMedCrossRefGoogle Scholar
  16. 16.
    Spink DC, Katz BH, Hussain MM, et al. (2003) Estrogen regulates Ah responsiveness in MCF-7 breast cancer cells. Carcinogenesis 24: 1941–1950.PubMedCrossRefGoogle Scholar
  17. 17.
    Payne J, Jones C, Lakhani S, et al. (2000) Improving the reproducibility of the MCF-7 cell proliferation assay for the detection of xenoestrogens. Sci Total Environ 248: 51–62.PubMedCrossRefGoogle Scholar
  18. 18.
    Soto AM, Sonnenschein C, Chung KL, et al. (1995) The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants. Environ Health Perspect 103 (Suppl 7): 113–122.PubMedCrossRefGoogle Scholar
  19. 19.
    Ricci MS, Toscano DG, Mattingly CJ, et al. (1999) Estrogen receptor reduces CYP1A1 induction in cultured human endometrial cells. J Biol Chem 274: 3430–3438.PubMedCrossRefGoogle Scholar
  20. 20.
    Safe S, Wormke M, Samudio I (2000) Mechanisms of inhibitory aryl hydrocarbon receptor-estrogen receptor crosstalk in human breast cancer cells. J Mammary Gland Biol Neoplasia 5: 295–306.PubMedCrossRefGoogle Scholar
  21. 21.
    Parvez S, Ismahan G, Raza-Bukhari A, et al. (1978) Activity of catechol-o-methyltransferase in brain regions and adrenal gland during the oestrus cycle. J Neural Transm 42: 305–312.PubMedCrossRefGoogle Scholar
  22. 22.
    Zhu BT, Liehr JG (1994) Quercetin increases the severity of estradiol-induced tumorigenesis in hamster kidney. Toxicol Appl Pharmacol 125: 149–158.PubMedCrossRefGoogle Scholar
  23. 23.
    Xie T, Ho SL, Ramsden D (1999) Characterization and implications of estrogenic down-regulation of human catechol-O-methyltransferase gene transcription. Mol Pharmacol 56: 31–38.PubMedGoogle Scholar
  24. 24.
    Rasmussen TH, Nielsen JB (2002) Critical parameters in the MCF-7 cell proliferation bioassay (E-Screen). Biomarkers 7: 322–336.PubMedCrossRefGoogle Scholar
  25. 25.
    Montano MM, Bianco NR, Deng H, et al. (2005) Estrogen receptor regulation of quinone reductase in breast cancer: implications for estrogen-induced breast tumor growth and therapeutic uses of tamoxifen. Front Biosci 10: 1440–1461.PubMedCrossRefGoogle Scholar
  26. 26.
    Lee JM, Anderson PC, Padgitt JK, et al. (2003) Nrf2, not the estrogen receptor, mediates catechol estrogen-induced activation of the antioxidant responsive element. Biochim Biophys Acta 1629: 92–101.PubMedGoogle Scholar
  27. 27.
    Setchell KD, Faughnan MS, Avades T, et al. (2003) Comparing the pharmacokinetics of daidzein and genistein with the use of 13C-labeled tracers in premenopausal women. Am J Clin Nutr 77: 411–419.PubMedGoogle Scholar
  28. 28.
    Kurzer MS (2002) Hormonal effects of soy in premenopausal women and men. J Nutr 132: 570S–573S.PubMedGoogle Scholar

Copyright information

© Springer 2008

Authors and Affiliations

  • Jörg Wagner
    • 1
  • Ling Jiang
  • Leane Lehmann
    • 2
  1. 1.Insitote of Applied Biosciences Section of Food Chemistry and ToxicologyUniversity of KarlsruheKarlsruheGermany
  2. 2.Institute of Applied BiosciencesUniversity of KarlsruheKarlsruheGermany

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