Assaying the estrogenicity of phytoestrogens in cells of different estrogen sensitive tissues.
There is currently much concern that a wide range of both synthetic and naturally occurring environmental chemicals may act as endocrine disruptors (ED), and may adversely affect humans and wildlife. We examined the estrogenic effects of the phytoestrogens daidzein (DAI), equol (EQU) and O-desmethylangolensin (O-DMA), two metabolites of DAI, in three different assays. Binding affinity to the estrogen receptor alpha was 1000-10,000-fold lower compared with the endogenous estrogen estradiol. In the receptor positive cell line MCF-7 the phytoestrogens induced the expression of a reporter gene. The E-SCREEN is based on the estrogen-receptor binding induced proliferation of the human breast cancer cell line MCF-7. We also adapted the E-SCREEN for the estrogen-receptor positive human ovarian cancer cell line BG-1. The tested phytoestrogens induced cell proliferation in both cell lines, but not in the receptor negative human breast cancer cell line MDA-MB-231. The phytoestrogen-induced cell proliferation could be blocked by addition of the receptor antagonist 4-hydroxytamoxifen (OHT). Combination treatments with the endogenous estrogen estradiol showed competitive effects in MCF-7 cells. These studies demonstrated that the tested phytoestrogens exerted estrogenic responses in cells derived from two different tissues, breast and ovary. Furthermore, we demonstrated that BG-1 cells are a suitable additional cell system to investigate estrogenicity of test compounds.
Schmitt
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Department of Toxicology, University of Wuerzburg, Versbacherstrasse 9, D-97078 Wuerzburg, Germany. eschmitt@toxi.uni-wuerzburg.de
Dekant
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Stopper
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D000826:Animal Testing Alternatives; D002455:Cell Division / Q000187:drug effects; D002839:Chromans / Q000494:pharmacology; D004336:Drug Antagonism; D060754:Equol; D004958:Estradiol / Q000494:pharmacology; D047628:Estrogen Receptor alpha; D004968:Estrogens, Non-Steroidal / Q000494:pharmacology*; D005260:Female; D017930:Genes, Reporter / Q000187:drug effects; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D048789:Phytoestrogens; D028321:Plant Preparations; D011960:Receptors, Estrogen / Q000378:metabolism*; D013629:Tamoxifen / Q000031:analogs & derivatives* / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000187:drug effects* / Q000378:metabolism
C007610:4',7-dihydroxy-3,4-dihydroisoflavone; D002839:Chromans; D047628:Estrogen Receptor alpha; D004968:Estrogens, Non-Steroidal; D007529:Isoflavones; D048789:Phytoestrogens; D028321:Plant Preparations; D011960:Receptors, Estrogen; D013629:Tamoxifen; C016601:afimoxifene; D004958:Estradiol; D060754:Equol; C004742:daidzein; D008156:Luciferases