Phytoestrogens Activate the Estrogen Receptor in HepG2 Cells

  • Lynne A. Kelly
Part of the Methods in Molecular Biology book series (MIMB, volume 1366)


Phytoestrogens are popular alternatives to estrogen therapy however their effects on hemostasis in postmenopausal women are unknown. This chapter describes a protocol to determine the effect of the phytoestrogens genistein, daidzein and equol, on the expression of key genes from the hemostatic system in human hepatocyte cell models and to determine the role of estrogen receptors in mediating any response seen using in vitro culture systems and Taqman® gene expression analysis.

Key words

Phytoestrogens Estrogenreceptor 17β-Estradiol Genistein Equol Daidzein HepG2 Hep89 


  1. 1.
    De Kleijn MJJ, van der Schouw YT, Wilson PWF et al (2002) Dietary intake of phytoestrogens is associated with a favorable metabolic cardiovascular risk profile in postmenopausal U.S.women: the Framingham study. J Nutr 132:276–282CrossRefGoogle Scholar
  2. 2.
    Kuiper GG, Lemmen JG, Carlsson B et al (1998) Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta. Endocrinology 139:4252–4263CrossRefGoogle Scholar
  3. 3.
    Pilšáková L, Riečanský I, Jagla F (2010) The physiological actions of isoflavone phytoestrogens. Physiol Res 59:651–664PubMedGoogle Scholar
  4. 4.
    Kelly LA, O’Leary JJ, Seidlova-Wuttke D et al (2010) Genistein alters coagulation gene expression in ovariectomised rats treated with phytoestrogens. Thromb Haemost 104:1250–1257CrossRefGoogle Scholar
  5. 5.
    Harnish DC, Evans MJ, Scicchitano MS et al (1998) Estrogen regulation of the apolipoprotein AI gene promoter through transcription cofactor sharing. J Biol Chem 273:9270–9278CrossRefGoogle Scholar
  6. 6.
    Norris LA, Brosnan J, Bonnar J et al (2008) Inhibitors and activation markers of the haemostatic system during hormone therapy: a comparative study of oral estradiol (2 mg)/dydrogesterone and estradiol (2 mg)/trimegestone. Thromb Haemost 100:253–260CrossRefGoogle Scholar
  7. 7.
    Lowe GD, Upton MN, Rumley A et al (2001) Different effects of oral and transdermal hormone replacement therapies on factor IX, APC resistance, t-PA, PAI and C-reactive protein—a cross-sectional population survey. Thromb Haemost 86:550–556CrossRefGoogle Scholar
  8. 8.
    Van Baal WM, Emeis JJ, van der Mooren MJ et al (2000) Impaired procoagulant-anticoagulant balance during hormone replacement therapy? A randomised, placebo-controlled 12-week study. Thromb Haemost 83:29–34CrossRefGoogle Scholar
  9. 9.
    Hulley S, Grady D, Bush T et al (1998) Randomized trial of estrogen plus progestin for secondary prevention of coronary heart disease in postmenopausal women. Heart and Estrogen/progestin Replacement Study (HERS) Research Group. JAMA 280:605–613CrossRefGoogle Scholar
  10. 10.
    Rossouw JE, Anderson GL, Prentice RL et al (2002) Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results From the Women’s Health Initiative randomized controlled trial. JAMA 288:321–333CrossRefGoogle Scholar
  11. 11.
    Kelly LA, Seidlova-Wuttke D, Wuttke W et al (2014) Estrogen receptor alpha augments changes in hemostatic gene expression in HepG2 cells treated with estradiol and phytoestrogens. Phytomedicine 21:155–158CrossRefGoogle Scholar
  12. 12.
    Farsetti A, Moretti F, Narducci M et al (1998) Orphan receptor hepatocyte nuclear factor-4 antagonizes estrogen receptor alpha-mediated induction of human coagulation factor XII gene. Endocrinology 139:4581–4589CrossRefGoogle Scholar
  13. 13.
    Farsetti A, Misiti S, Citarella F et al (1995) Molecular basis of estrogen regulation of Hageman factor XII gene expression. Endocrinology 136:5076–5083CrossRefGoogle Scholar
  14. 14.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408CrossRefGoogle Scholar
  15. 15.
    Berthois Y, Katzenellenbogen JA, Katzenellenbogen BS (1986) Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci U S A 83:2496–2500CrossRefGoogle Scholar
  16. 16.
    Mann GE, Bonacasa B, Ishii T, Siow RC (2009) Targeting the redox sensitive Nrf2-Keap1 defense pathway in cardiovascular disease: protection afforded by dietary isoflavones. Curr Opin Pharmacol 9:139–145CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of PaediatricsThe Coombe Women and Infants University Hospital & Trinity CollegeDublin 8Ireland

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