Development of a stable cell line with an intact PGC-1α/ERRα axis for screening environmental chemicals.
The estrogen-related receptor α (ERRα) and the peroxisome proliferator-activated receptor γ (PPARγ) coactivator 1α (PGC-1α) play critical roles in the control of several physiological functions, including the regulation of genes involved in energy homeostasis. However, little is known about the ability of environmental chemicals to disrupt or modulate this important bioenergetics pathway in humans. The goal of this study was to develop a cell-based assay system with an intact PGC-1α/ERRα axis that could be used as a screening assay for detecting such chemicals. To this end, we successfully generated several stable cell lines expressing PGC-1α and showed that the reporter driven by the native ERRα hormone response unit (AAB-Luc) is active in these cell lines and that the activation is PGC-1α-dependent. Furthermore, we show that this activation can be blocked by the ERRα selective inverse agonist, XCT790. In addition, we find that genistein and bisphenol A further stimulate the reporter activity, while kaempferol has minimal effect. These cell lines will be useful for identifying environmental chemicals that modulate this important pathway.
DNTP, BioMolecular Screening Branch, Division, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, United States. Electronic address: teng1@niehs.nih.gov.
Christina T
CT
Teng
DNTP, BioMolecular Screening Branch, Division, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, United States.
Burton
B
Beames
DNTP, BioMolecular Screening Branch, Division, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, United States.
B
B
Alex Merrick
DIR, Viral Core Lab, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, United States.
Negin
N
Martin
DIR, Viral Core Lab, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, United States.
Charles
C
Romeo
DIR Laboratory of Respiratory Biology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, United States.
Anton M
AM
Jetten
D000395:Air Pollutants, Occupational / Q000494:pharmacology; D001559:Benzhydryl Compounds / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods; D015153:Blotting, Western; D019833:Genistein / Q000494:pharmacology; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D010636:Phenols / Q000494:pharmacology; D048789:Phytoestrogens / Q000494:pharmacology; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics*; D013844:Thiazoles / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D015262:Xenobiotics / Q000494:pharmacology*; D000097983:ERRalpha Estrogen-Related Receptor; D000098826:Bisphenol A Compounds
D000395:Air Pollutants, Occupational; D001559:Benzhydryl Compounds; D019833:Genistein; D008156:Luciferases; D009570:Nitriles; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D010636:Phenols; D048789:Phytoestrogens; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D013844:Thiazoles; D014157:Transcription Factors; D015262:Xenobiotics; D000097983:ERRalpha Estrogen-Related Receptor; C486497:PPARGC1A protein, human; C488234:XCT790; C006780:bisphenol A; D000098826:Bisphenol A Compounds
EDC; Luc reporter; PGC-1α/ERRα; Screen; Stable cell line; XCT790