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Endocrinology Vol. 139, No. 10 4252-4263
Copyright © 1998 by The Endocrine Society


ARTICLES

Interaction of Estrogenic Chemicals and Phytoestrogens with Estrogen Receptor ß

George G. J. M. Kuiper1, Josephine G. Lemmen, Bo Carlsson, J. Christopher Corton, Stephen H. Safe, Paul T. van der Saag, Bart van der Burg2 and Jan-Åke Gustafsson3

Center for Biotechnology and Department of Medical Nutrition (G.G.J.M.K., J.-Å.G.), Karolinska Institute and KaroBio AB (B.C.) Huddinge, Sweden; Hubrecht Laboratory, Netherlands Institute for Developmental Biology (B.v.d.B., P.T.v.d.S., J.G.L.) Utrecht, The Netherlands; Chemical Industry Institute of Toxicology (J.C.C.), Research Triangle Park, North Carolina; Department of Veterinary Physiology and Pharmacology (S.H.S.), Texas A&M University, College Station, Texas 77843-4466

Address all correspondence and requests for reprints to: Dr. George Kuiper, Center for Biotechnology, NOVUM, S-14186 Huddinge, Sweden. E-mail: george.kuiper{at}csb.ki.se

The rat, mouse and human estrogen receptor (ER) exists as two subtypes, ER{alpha} and ERß, which differ in the C-terminal ligand-binding domain and in the N-terminal transactivation domain. In this study, we investigated the estrogenic activity of environmental chemicals and phytoestrogens in competition binding assays with ER{alpha} or ERß protein, and in a transient gene expression assay using cells in which an acute estrogenic response is created by cotransfecting cultures with recombinant human ER{alpha} or ERß complementary DNA (cDNA) in the presence of an estrogen-dependent reporter plasmid.

Saturation ligand-binding analysis of human ER{alpha} and ERß protein revealed a single binding component for [3H]-17ß-estradiol (E2) with high affinity [dissociation constant (Kd) = 0.05 - 0.1 nM]. All environmental estrogenic chemicals [polychlorinated hydroxybiphenyls, dichlorodiphenyltrichloroethane (DDT) and derivatives, alkylphenols, bisphenol A, methoxychlor and chlordecone] compete with E2 for binding to both ER subtypes with a similar preference and degree. In most instances the relative binding affinities (RBA) are at least 1000-fold lower than that of E2. Some phytoestrogens such as coumestrol, genistein, apigenin, naringenin, and kaempferol compete stronger with E2 for binding to ERß than to ER{alpha}. Estrogenic chemicals, as for instance nonylphenol, bisphenol A, o, p'-DDT and 2',4',6'-trichloro-4-biphenylol stimulate the transcriptional activity of ER{alpha} and ERß at concentrations of 100-1000 nM. Phytoestrogens, including genistein, coumestrol and zearalenone stimulate the transcriptional activity of both ER subtypes at concentrations of 1–10 nM. The ranking of the estrogenic potency of phytoestrogens for both ER subtypes in the transactivation assay is different; that is, E2 >> zearalenone = coumestrol > genistein > daidzein > apigenin = phloretin > biochanin A = kaempferol = naringenin > formononetin = ipriflavone = quercetin = chrysin for ER{alpha} and E2 >> genistein = coumestrol > zearalenone > daidzein > biochanin A = apigenin = kaempferol = naringenin > phloretin = quercetin = ipriflavone = formononetin = chrysin for ERß. Antiestrogenic activity of the phytoestrogens could not be detected, except for zearalenone which is a full agonist for ER{alpha} and a mixed agonist-antagonist for ERß. In summary, while the estrogenic potency of industrial-derived estrogenic chemicals is very limited, the estrogenic potency of phytoestrogens is significant, especially for ERß, and they may trigger many of the biological responses that are evoked by the physiological estrogens.




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