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Comparison of short-term estrogenicity tests for identification of hormone-disrupting chemicals.

Environmental health perspectives (1999-05-07)
H R Andersen, A M Andersson, S F Arnold, H Autrup, M Barfoed, N A Beresford, P Bjerregaard, L B Christiansen, B Gissel, R Hummel, E B Jørgensen, B Korsgaard, R Le Guevel, H Leffers, J McLachlan, A Møller, J B Nielsen, N Olea, A Oles-Karasko, F Pakdel, K L Pedersen, P Perez, N E Skakkeboek, C Sonnenschein, A M Soto
ABSTRACT

The aim of this study was to compare results obtained by eight different short-term assays of estrogenlike actions of chemicals conducted in 10 different laboratories in five countries. Twenty chemicals were selected to represent direct-acting estrogens, compounds with estrogenic metabolites, estrogenic antagonists, and a known cytotoxic agent. Also included in the test panel were 17beta++-estradiol as a positive control and ethanol as solvent control. The test compounds were coded before distribution. Test methods included direct binding to the estrogen receptor (ER), proliferation of MCF-7 cells, transient reporter gene expression in MCF-7 cells, reporter gene expression in yeast strains stably transfected with the human ER and an estrogen-responsive reporter gene, and vitellogenin production in juvenile rainbow trout. 17beta-Estradiol, 17alpha-ethynyl estradiol, and diethylstilbestrol induced a strong estrogenic response in all test systems. Colchicine caused cytotoxicity only. Bisphenol A induced an estrogenic response in all assays. The results obtained for the remaining test compounds--tamoxifen, ICI 182.780, testosterone, bisphenol A dimethacrylate, 4-n-octylphenol, 4-n-nonylphenol, nonylphenol dodecylethoxylate, butylbenzylphthalate, dibutylphthalate, methoxychlor, o,p'-DDT, p,p'-DDE, endosulfan, chlomequat chloride, and ethanol--varied among the assays. The results demonstrate that careful standardization is necessary to obtain a reasonable degree of reproducibility. Also, similar methods vary in their sensitivity to estrogenic compounds. Thus, short-term tests are useful for screening purposes, but the methods must be further validated by additional interlaboratory and interassay comparisons to document the reliability of the methods.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
β-Estradiol, analytical standard