Evaluation of In vitro and In vivo Screening Methods for Estrogenic Activity of Endocrine Disruptors

내분비계 장애물질의 에스트로겐 활성에 대한 In vitro 및 In vivo 검색시험법

  • 김형식 (국립독성연구소 독성부) ;
  • 한순영 (국립독성연구소 독성부) ;
  • 한상국 (국립독성연구소 독성부) ;
  • 신재호 (국립독성연구소 독성부) ;
  • 문현주 (국립독성연구소 독성부) ;
  • 김소희 (국립독성연구소 독성부) ;
  • 박기숙 (국립독성연구소 독성부) ;
  • 김규봉 (국립독성연구소 독성부) ;
  • 이이다 (국립독성연구소 독성부)
  • Published : 2000.06.01

Abstract

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. 17$\beta$-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 $\mu\textrm{g}$/kg) and EE (0.3 $\mu\textrm{g}$/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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