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KMID : 0379520000160020109
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한국독성학회지
2000 Volume.16 No. 2 p.109 ~ p.116
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Evaluation of In vitro and In vivo Screening Methods for Estrogenic Activity of Endocrine Disruptors
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Kim Hyoung-Sik
Han Soon-Young
Han Sang-Kuk
Shin Jae-Hoo
Moon Hyun-Ju
Kim So-Hee
Park Ki-Sook
Kim Kyu-Bong
Lee Rhee-Da
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Abstract
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The purposes of our study were to optimize the conditions of the screening and testing methods for endocrine disruptors, to characterize these assays using several compounds with well-defined endocrine activity, and to compare the sensitivity between these assays currently undergoing validation. Two in vitro test systems, MCF-7 cells proliferation (E-screen assay) and competitive binding to estrogen receptors (ER) were selected to evaluate the estrogenic effects. 17beta-Estradiol (E2) and diethylstilbestrol (DES) were used as a positive control in vitro test. Also, E2 and ethinyl estradiol (EE) were used as a positive control in vivo uterotrophic assay. In in vitro test, E2 and DES showed a strong estrogenic response at concentration of 1.0 nM. In uterotrophic assay, E2 (0.3 mutextrm{g}/kg) and EE (0.3 mutextrm{g}/kg) produced a significant increase in uterus and vagina weight in both immature and ovariectomized rats. Although we did not com-pared the specificity between in vivo and in vitro assays, these assay systems may serve as a good tool for endocrine disruptors screening methods. Our data indicate that these assay systems exhibit some difference in their sensitivity to the same estrogenic compounds. Therefore, as a first rapid screening assay for estrogenic activity qf unknown chemicals, at least two assay systems should probably be carried out with a view of high sensitivity and standardization conditions. Also, a careful validation tests are necessary to obtain a reasonable degree of reproducibility.
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KEYWORD
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Endocrine disruptors, MCF-7 cells, E-screen assay, Competitive binding, Uterotrophic assay
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