Detection of thyroid hormone receptor disruptors by a novel stable in vitro reporter gene assay

Jaime Freitas, Patricia Cano, Christina Craig-Veit, Michael L. Goodson, John Furlow, Albertinka J. Murk

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

A stable luciferase reporter gene assay was developed based on the thyroid hormone responsive rat pituitary tumor GH3 cell line that constitutively expresses both thyroid hormone receptor isoforms. Stable transfection of the pGL4CP-SV40-2xtaDR4 construct into the GH3 cells resulted in a highly sensitive cell line (GH3.TRE-Luc), which was further optimized into an assay that allowed the detection of Triiodothyronine (T3) and Thyroxine (T4) concentrations in the picomolar range after only 24h of exposure. The greater than 20-fold induction of T3 relative to the solvent control is illustrative of the high responsiveness of the system. The assay was validated by the quantification of the agonistic effect of the natural hormones (T3 and T4), the acetic acid derivatives of T3 (triiodothyroaceticacid, or Triac) and T4 (tetraiodothyroacetic acid, or Tetrac), hydroxy polybrominated diphenylethers (OH-PBDEs), hydroxy polychlorinated biphenyls (OH-PCBs) and the antagonistic action of sodium arsenite (NaAsO2). The putative antagonist Amiodarone, Bisphenol A (BPA) and its halogenated derivatives (TCBPA and TBBPA) for which effects reported in the literature are not consistent, showed comparable dose-response curves with a slight agonistic effect (5% of T3-max) followed by a slight antagonistic effect. The magnitude and reproducibility of the responses to various compounds confirms this assay as a promising tool for the identification and quantification of specific thyroid hormone receptor disrupting potency of compounds.

Original languageEnglish (US)
Pages (from-to)257-266
Number of pages10
JournalToxicology in Vitro
Volume25
Issue number1
DOIs
StatePublished - Feb 1 2011

Fingerprint

Thyroid Hormone Receptors
Reporter Genes
Assays
Genes
Amiodarone
Polychlorinated Biphenyls
Pituitary Neoplasms
Triiodothyronine
Tumor Cell Line
Luciferases
Thyroxine
Thyroid Hormones
Acetic Acid
Transfection
Protein Isoforms
Cells
Hormones
Derivatives
Cell Line
Rats

Keywords

  • Amiodarone
  • Bisphenol A
  • Endocrine disruption
  • PBDE
  • PCB metabolite
  • Thyroid hormone receptor

ASJC Scopus subject areas

  • Toxicology

Cite this

Detection of thyroid hormone receptor disruptors by a novel stable in vitro reporter gene assay. / Freitas, Jaime; Cano, Patricia; Craig-Veit, Christina; Goodson, Michael L.; Furlow, John; Murk, Albertinka J.

In: Toxicology in Vitro, Vol. 25, No. 1, 01.02.2011, p. 257-266.

Research output: Contribution to journalArticle

Freitas, Jaime ; Cano, Patricia ; Craig-Veit, Christina ; Goodson, Michael L. ; Furlow, John ; Murk, Albertinka J. / Detection of thyroid hormone receptor disruptors by a novel stable in vitro reporter gene assay. In: Toxicology in Vitro. 2011 ; Vol. 25, No. 1. pp. 257-266.
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