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Comparison of reporter gene assay and immature rat uterotrophic assay of twenty-three chemicals.

Abstract：

:We performed a reporter gene assay for ERalpha-mediated transcriptional activation and an immature rat uterotrophic assay of 23 chemicals, to study the relationship between these two assays and to examine the usefulness of the reporter gene assay. The chemicals analyzed in the study were as follows: benzophenone, bisphenol A, bisphenol B, bisphenol F, p-cumyl phenol, dibutyl phthalate, dicyclohexylphthalate, dihydrotestosterone, equilin, 17alpha-estradiol, estrone, ethynyl estradiol, genistein, hematoxylin, nonylphenol mixture, 4-n-nonylphenol, norethindrone, norgestrel, octachlorostyrene, 4-n-octylphenol, 4-tert-octylphenol, tributyltin-chloride and zearalenone. To perform the reporter gene assay, HeLa cells were transfected with a rat ERalpha expression construct and an estrogen-regulated luciferase reporter construct. The transcriptional activities of each chemical were tested over concentrations ranging from 10 pM to 10 microM and the EC50, PC50 and PC10 values were calculated. In the immature rat uterotrophic assay, the doses of 21 chemicals, with the exception of dibutyl phthalate and ethynyl estradiol, were 0, 2, 20 and 200 mg/kg; each group consisted of six rats. The doses of dibutyl phthalate and ethynyl estradiol were 0, 40, 200 and 1000 mg/kg per day and 0, 0.2, 2 and 20 microg/kg per day, respectively. In the reporter gene assay, the PC10 values were calculated for 15 chemicals: bisphenol A, bisphenol B, bisphenol F, p-cumyl phenol, dihydrotestosterone, equilin, 17alpha-estradiol, estrone, ethynyl estradiol, genistein, nonylphenol mixture, norethindrone, norgestrel, 4-tert-octylphenol and zearalenone. These chemicals corresponded to the chemicals that tested positive in the uterotrophic assay. The other chemicals were negative in the reporter and uterotrophic assays. Although the EC50 and PC50 values could only be calculated for five and six chemicals, respectively, the PC10 values were shown to be well correlated with the EC50 values by a correlation analysis (R(2)=0.9202). These findings demonstrate that PC10 values are preferable to EC50 and PC50 values for predicting the estrogenic activities of chemicals.

journal_name

Toxicology

journal_title

Toxicology

authors

Yamasaki K,Takeyoshi M,Yakabe Y,Sawaki M,Imatanaka N,Takatsuki M

doi

10.1016/s0300-483x(01)00505-4

keywords:

subject

Has Abstract

pub_date

2002-01-15 00:00:00

pages

21-30

issue

1-2

eissn

0300-483X

issn

1879-3185

pii

S0300483X01005054

journal_volume

170

pub_type

杂志文章

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Comparison of reporter gene assay and immature rat uterotrophic assay of twenty-three chemicals.