Archives of Pharmacal Research

, Volume 26, Issue 9, pp 756–762 | Cite as

Assessment of the estrogenicity of isoflavonoids, using MCF-7-ERE-Luc cells

  • Ki Eun Joung
  • Yeo Woon Kim
  • Yhun Yhong Sheen


In the current study, our research focused on the estrogenic activity of isoflavonoids, mainly genistein, biochanin A and daidzein. Genistein enhanced the reporter gene expression of MCF-7-ERE-Luc cells, at a concentration as low as 10 nM, with a concentration of 100 nM the achieved gene expression effects were similar to those of 10 pM 17β-estradiol. Based on the estrogenic activities of biochanin A and daidzein, hydroxyl groups at the 4′ and 5 positions are needed for the maximal effect of the genistein. The estrogenic effects of these isoflavonoids were inhibited by the concomitant treatment with tamoxifen. The data showed that the estrogenic effects of isoflavonoids were mediated through estrogen receptors. When the isoflavonoids were tested as mixtures, the estrogenic effects were lower than the arithmetic sum of those induced by each individual isoflavonoid. The estrogenic potency of each isoflavonoid was presented at EC50 levels with a 17β-estradiol equivalent concentration (EEQ) based on the dose response of each chemical. The EC50s and EEQs of genistein, biochanin A and daidzein were 4.15, 0.89 and 0.18 μM, and 15.0, 5.12 and 1.83 μM/M, respectively. Our data clearly demonstrated that the pERE-luciferase reporter gene assay was suited for the sensitive and quantitative measurement, and large scale screening, of the estrogenicity of chemicalsin vitro.

Key words

Genistein Biochanin A Daidzein EEQ 17β-Estradiol ERE 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Andersen, H. R., Andersson, A. M., Arnold, S. F., Autrup, H., Barfoed, M., Beresford, N.A., Bjerregaard, P., Christiansen, L. B., Gissel, B., Hummel, R., Jorgensen, E. B., Korsgaard, B., Le Guevel, R., Leffers, H., McLachlan, J., Moller, A., Nielsen, J. B., Olea, N., Oles-Karasko, A., Pakdel, F., Pedersen, K. L., Perez, P., Skakkeboek, N. E., Sonnenschein, C., and Soto, A. al., Comparison of short-term estrogenicity tests for identification of hormone-disrupting chemicals.Environ, Health Perspect, 107, 89–108 (1999).CrossRefGoogle Scholar
  2. Andersson, A. M. and Skakkebaek, N. E., Exposure to exogenous estrogens in food: possible impact on human development and health.Eur. J. Endocrinol., 140, 477–485 (1999).PubMedCrossRefGoogle Scholar
  3. Ansbacher, R., Selective estrogen receptor modulators.Am. J. Obstet. Gynecol., 181, 1036 (1999).PubMedCrossRefGoogle Scholar
  4. Bolger, R., Wiese, T. E., Ervin, K., Nestich, S., and Checovich, W., Rapid screening of environmental chemicals for estrogen receptor binding capacity.Environ. Health Prespect, 106, 551–557 (1998).CrossRefGoogle Scholar
  5. Breinholt, V. and Larsen, J. C., Detection of weak estrogenic flavonoids using a recombinant yeast strain and a modified MCP7 cell proliferation assay.Chem. Res. Toxicol., 11, 622–629 (1998).PubMedCrossRefGoogle Scholar
  6. Brzezinski, A. and Debi, A., Phytoestrogens: the “natural” selective estrogen receptor modulators?Eur. J. Obstet. Gynecol. Reprod. Biol., 85, 47–51 (1999).PubMedCrossRefGoogle Scholar
  7. Cassidy, A., Bingham, S., and Setchell K. D., Biological effects of a diet of soy protein rich in isoflavones on the menstrual cycle of premenopausal women.Am. J. Clin. Nutr., 60, 333–340 (1994).PubMedGoogle Scholar
  8. Choi, Y. H., Zhang, L., Lee, W. H., and Park, K. Y., Genistein-induced G2/M arrest is associated with the inhibition of cyclin B1 and the induction of p21 in human breast carcinoma cells.Int. J. Oncol., 3, 391–396 (1998).Google Scholar
  9. Constantinou, A. I., Kamath, N., and Murley, J. S., Genistein inactivates bcl-2, delays the G2/M phase of the cell cycle, and induces apoptosis of human breast adenocarcinoma MCR-7 cells.Eur. J. Cancer, 34, 1927–1934 (1998)PubMedCrossRefGoogle Scholar
  10. Delmas, P. D., Clinical use of selective estrogen receptor modulators.Bone, 25, 115–118 (1999).PubMedCrossRefGoogle Scholar
  11. Rioravanti, L., Cappelletti, V., Miodini, P., Ronchi, E., Brivio, M., and Di Fronzo, G., Genistein in the control of breast cancer cell growth: insights into the mechanism of actionin vitro.Cancer Lett., 130, 143–152 (1998).CrossRefGoogle Scholar
  12. Potsis, T., Pepper, M. S., Aktas, E., Breit, S., Rasku, S., Adlercreutz, H., Wahala, K., Montesano, R., and Schweigerer, L., Flavonoids, dietary-derived inhibitors of cell proliferation andin vitro angiogenesis.Cancer Res., 57, 2916–2921 (1997).Google Scholar
  13. Gaido, K. W., Leonard, L. S., Lovell, S., Gould, J. C., Babai, D., Portier, C. J., and MacDonell, D. P., Evaluation of chemicals with endocrine modulating activity in a yeast-based steroid hormone receptor transcription assay.Toxicol. Appl. Pharmacol., 143, 205–212 (1997).PubMedCrossRefGoogle Scholar
  14. Hochner-Celnikier, D., Pharmacokinetics of raloxifene and its clinical applications.Eur. J. Obstet. Gynecol. Reprod. Biol., 85, 47–51 (1999).CrossRefGoogle Scholar
  15. Jorgensen, M., Vendelbo, B., Skakkebaek, N. E., and Leffers, H., Assaying estrogenicity by quantitating the expression of endogenous estrogen-regulated genes.Environ. Health Perspect, 108, 403–412 (2000).PubMedCrossRefGoogle Scholar
  16. Klein, K. O., Baron, J., Colli, M. J., McDonnell, D. P., and Cutler, G. B. Jr., Estrogen levels in childhood determined by an ultrasensitive recombinant cell bioassay.J. Clin. Invest., 94, 2475–2480 (1994).PubMedCrossRefGoogle Scholar
  17. Kuzumaki, T., Kobayashi, T., and Ishikawa, K., Genistein induces p21(Cip1/WAP1) expression and blocks the G1 to S phase transition in mouse fibroblast and melanoma cells.Biochem. Biophys. Res. Commun., 251, 291–295 (1998).PubMedCrossRefGoogle Scholar
  18. Masamura, S., Santner, S. J., Heitjan, D. P., and Santen, R. J., Estrogen deprivation causes estradiol hypersensitivitiy in human breast cancer cells.J. Clin. Endocrinol. Metab., 80, 2918–2925 (1995).PubMedCrossRefGoogle Scholar
  19. Meyer, T., Koop, R., Von Angerer, E., Schonenberger, H., and Holler, E., A rapid luciferase transfection assay for transcription activation effects and stability control of estrogenic drugs in cell cultures.J. Cancer Res. Clin. Oncol., 120, 359–364 (1994).PubMedCrossRefGoogle Scholar
  20. Morito, K., Aomori T., Hirose, T., Kinjo J., Hasegawa, J., Ogawa, S., Inoue, S., Muramatsu, M., and Masamune, Y., Interaction of Phytoestrogens with Estrogen Receptors α and β (II).Biol. Pharm. Bull., 25, 48–52 (2002).PubMedCrossRefGoogle Scholar
  21. Nishikawa, J., Saito, K., Goto, J., Dakeyama, F., Matsuo, M., and Nishihara, T., New screening methods for chemicals with hormonal activities using interaction of nuclear hormone receptor with coactivator.Toxico. Appl. Pharmacol., 154, 76–83 (1999).CrossRefGoogle Scholar
  22. Noroozi, M., Angerson, W. J., and Lean, M. E., Effects of flavonoids and vitamin C on oxidative DNA damage to human lymphocytes.Am. J. Clin. Nutr., 67, 1210–1218 (1998).PubMedGoogle Scholar
  23. Soto, A. M., Sonnenschein, C., Chung, K. L., Fernandes, M. F., Olea, N., and Serrano, F. O. The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants.Environ. Health Perspect, 103, 113–122 (1995).PubMedCrossRefGoogle Scholar
  24. Sumpter, J. P. and Jobling, S., Vitellogenesis as a biomarker for estrogenic contamination of the aquatic environment.Environ. Health Perspect, 103, 173–178 (1995).PubMedCrossRefGoogle Scholar
  25. Wang, T. T., Sathyamoorthy, N., and Phang, J. M., Molecular effects of genistein on estrogen receptor mediated pathways.Carcinogenesis, 17, 271–275 (1996).PubMedCrossRefGoogle Scholar
  26. Weryha, G., Pascal-Vigneron, V., Klein, M., and Leclere, J., Selective estrogen receptor modulators.Curr. Opin. Rheumatol., 11, 301–306 (1999).PubMedCrossRefGoogle Scholar
  27. Xu, X., Duncan, A. M., Merz, B. E., and Kurzer, M. S., Effects of soy isoflavones on estrogen and phytoestrogen metabolism in premenopausal women.Cancer Epidemiol. Biomarkers Prev., 7, 1101–1108 (1998).PubMedGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2003

Authors and Affiliations

  • Ki Eun Joung
    • 1
  • Yeo Woon Kim
    • 1
  • Yhun Yhong Sheen
    • 1
  1. 1.College of PharmacyEwha Womans UniversitySeodaemun-Ku, SeoulKorea

Personalised recommendations