Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor-reporter gene assays

Patrick Balaguer, Annick Joyeux, Michael S. Denison, Renaud Vincent, Bradley E. Gillesby, Tim Zacharewski

Research output: Contribution to journalArticle

72 Scopus citations

Abstract

In vitro recombinant receptor-reporter gene assays have been used to assess and rank the potency of chemicals and complex mixtures suspected of possessing estrogen and (or) aryl hydrocarbon receptor (AhR) mediated activity. The environmental estrogen (E2) bioassay consists of a Gal4-human estrogen receptor chimeric construct (Gal4-HEGO) and a Gal4-regulated luciferase reporter gene (17m5-G-Luc) that have been stably integrated into HeLa cells. The assay exhibits 10-fold induction in luciferase reporter gene activity following treatment with 1 nM 17β-estradiol and has a detection limit of approximately 5 pg of 17β-estradiol/mL. The AhR bioassay uses Hepa 1c1c7 wild-type cells transiently transfected with a dioxin response element regulated luciferase reporter gene. These assays were used to assess the estrogen and dioxin-like activities of naringenin, atrazine, and simazine and complex mixtures such as pulp and paper mill black liquor and urban air particulates. The activities of these chemicals and complex mixtures are confirmed using the pure antiestrogen ICI 164,384 and in in vitro gel retardation assays. Results of this study demonstrate the utility of in vitro recombinant receptor-reporter gene assays in identifying and assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures.

Original languageEnglish (US)
Pages (from-to)216-222
Number of pages7
JournalCanadian Journal of Physiology and Pharmacology
Volume74
Issue number2
DOIs
StatePublished - 1996

Keywords

  • bioassays
  • dioxin-like
  • environmental estrogen
  • in vitro
  • receptor

ASJC Scopus subject areas

  • Physiology
  • Pharmacology

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