Development of a recombinant human ovarian (BG1) cell line containing estrogen receptor α and β for improved detection of estrogenic/antiestrogenic chemicals

Environ Toxicol Chem. 2016 Jan;35(1):91-100. doi: 10.1002/etc.3146. Epub 2015 Dec 9.

Abstract

Estrogenic endocrine-disrupting chemicals are found in environmental and biological samples, commercial and consumer products, food, and numerous other sources. Given their ubiquitous nature and potential for adverse effects, a critical need exists for rapidly detecting these chemicals. The authors developed an estrogen-responsive recombinant human ovarian (BG1Luc4E2) cell line recently accepted by the US Environmental Protection Agency (USEPA) and Organisation for Economic Co-operation and Development (OECD) as a bioanalytical method to detect estrogen receptor (ER) agonists/antagonists. Unfortunately, these cells appear to contain only 1 of the 2 known ER isoforms, ERα but not ERβ, and the differential ligand selectivity of these ERs indicates that the currently accepted screening method only detects a subset of total estrogenic chemicals. To improve the estrogen screening bioassay, BG1Luc4E2 cells were stably transfected with an ERβ expression plasmid and positive clones identified using ERβ-selective ligands (genistein and Br-ERβ-041). A highly responsive clone (BG1LucERβc9) was identified that exhibited greater sensitivity and responsiveness to ERβ-selective ligands than BG1Luc4E2 cells, and quantitative reverse-transcription polymerase chain reaction confirmed the presence of ERβ expression in these cells. Screening of pesticides and industrial chemicals identified chemicals that preferentially stimulated ERβ-dependent reporter gene expression. Together, these results not only demonstrate the utility of this dual-ER recombinant cell line for detecting a broader range of estrogenic chemicals than the current BG1Luc4E2 cell line, but screening with both cell lines allows identification of ERα- and ERβ-selective chemicals.

Keywords: BG1Luc ER TA; Environmental screening; Estrogen receptor bioassay; Estrogenic chemicals.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay
  • Cell Line
  • Endocrine Disruptors / toxicity*
  • Environmental Pollutants / toxicity*
  • Estrogen Antagonists / toxicity*
  • Estrogen Receptor alpha / drug effects*
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / metabolism*
  • Estrogen Receptor beta / drug effects*
  • Estrogen Receptor beta / genetics
  • Estrogen Receptor beta / metabolism*
  • Estrogens, Non-Steroidal / toxicity*
  • Female
  • Gene Expression / drug effects
  • Genistein / pharmacology
  • Humans
  • Ovary / cytology
  • Ovary / metabolism*
  • Pesticides / toxicity
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Transfection

Substances

  • ESR1 protein, human
  • Endocrine Disruptors
  • Environmental Pollutants
  • Estrogen Antagonists
  • Estrogen Receptor alpha
  • Estrogen Receptor beta
  • Estrogens, Non-Steroidal
  • Pesticides
  • Genistein