Assaying estrogenicity by quantitating the expression levels of endogenous estrogen-regulated genes

M Jørgensen, B Vendelbo, N E Skakkebaek, H Leffers

Abstract

Scientific evidence suggests that humans and wildlife species may experience adverse health consequences from exposure to environmental chemicals that interact with the endocrine system. Reliable short-term assays are needed to identify hormone-disrupting chemicals. In this study we demonstrate that the estrogenic activity of a chemical can be evaluated by assaying induction or repression of endogenous estrogen-regulated "marker genes" in human breast cancer MCF-7 cells. We included four marker genes in the assay--pS2, transforming growth factor beta3 (TGFbeta3), monoamine oxidase A, and [alpha]1-antichymotrypsin--and we evaluated estrogenic activity for 17beta-estradiol (E(2)), diethylstilbestrol, [alpha]-zearalanol, nonylphenol, genistein, methoxychlor, endosulphan, o,p-DDE, bisphenol A, dibutylphthalate, 4-hydroxy tamoxifen, and ICI 182.780. All four marker genes responded strongly to the three high-potency estrogens (E(2), diethylstilbestrol, and [alpha]-zearalanol), whereas the potency of the other chemicals was 10(3)- to 10(6)-fold lower than that of E(2). There were some marker gene-dependent differences in the relative potencies of the tested chemicals. TGFbeta3 was equally sensitive to the three high-potency estrogens, whereas the sensitivity to [alpha]-zearalanol was approximately 10-fold lower than the sensitivity to E(2) and diethylstilbestrol when assayed with the other three marker genes. The potency of nonylphenol was equal to that of genistein when assayed with pS2 and TGFbeta3, but 10- to 100-fold higher/lower with monoamine oxidase A and [alpha]1-antichymotrypsin, respectively. The results are in agreement with results obtained by other methods and suggest that an assay based on endogenous gene expression may offer an attractive alternative to other E-SCREEN methods.

Original languageEnglish
JournalEnvironmental Health Perspectives
Volume108
Issue number5
Pages (from-to)403-12
Number of pages10
ISSN0091-6765
DOIs
Publication statusPublished - May 2000

Keywords

  • Animals
  • Base Sequence
  • Breast Neoplasms/genetics
  • DNA Primers/genetics
  • Environmental Health
  • Environmental Pollutants/toxicity
  • Estrogen Receptor Modulators/pharmacology
  • Estrogens/genetics
  • Estrogens, Non-Steroidal/toxicity
  • Female
  • Gene Expression/drug effects
  • Genetic Markers
  • Humans
  • Monoamine Oxidase/genetics
  • Proteins/genetics
  • RNA, Messenger/biosynthesis
  • Transforming Growth Factor beta/genetics
  • Trefoil Factor-1
  • Tumor Cells, Cultured
  • Tumor Suppressor Proteins
  • alpha 1-Antichymotrypsin/genetics

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