Novel stably transfected gene reporter human hepatoma cell line for assessment of aryl hydrocarbon receptor transcriptional activity: construction and characterization
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
22017252
DOI
10.1021/es2029334
Knihovny.cz E-resources
- MeSH
- Anthracenes pharmacology MeSH
- Hep G2 Cells MeSH
- Gene Expression drug effects genetics MeSH
- Indoles pharmacology MeSH
- Humans MeSH
- Luciferases genetics metabolism MeSH
- Methylcholanthrene pharmacology MeSH
- Omeprazole pharmacology MeSH
- Polychlorinated Dibenzodioxins pharmacology MeSH
- Receptors, Aryl Hydrocarbon genetics metabolism MeSH
- Resveratrol MeSH
- Signal Transduction MeSH
- Stilbenes pharmacology MeSH
- Transfection MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Anthracenes MeSH
- indirubin MeSH Browser
- Indoles MeSH
- Luciferases MeSH
- Methylcholanthrene MeSH
- Omeprazole MeSH
- Polychlorinated Dibenzodioxins MeSH
- pyrazolanthrone MeSH Browser
- Receptors, Aryl Hydrocarbon MeSH
- Resveratrol MeSH
- Stilbenes MeSH
We constructed stably transfected gene reporter cell line AZ-AHR, allowing measurement of aryl hydrocarbon receptor (AhR) transcriptional activity. Human hepatoma HepG2 cells were transfected with a construct containing several AhR binding sites upstream of luciferase reporter gene. We prepared 12 clones and we characterized the best five in responsiveness to TCDD. Dose-response analyses were performed for various AhR ligands, including TCDD, 3-methylcholanthrene, indirubin, resveratrol, omeprazole, and SP600125. The EC(50) values were similar in all tested clones. Induction of luciferase was time-dependent, and treatment for 6 h with 5 nM TCDD was sufficient to evaluate AhR transcriptional activity in 96-well plate format (8-24 fold induction). Response to AhR ligands of cryopreserved cells after thawing was not significantly different from that of fresh cells. Cell line remained fully responsive to AhR ligands over 15 passages and 30 days in culture without significant alterations. Overall, we have developed novel human luciferase reporter cell line AZ-AHR for monitoring AhR transcriptional activity. The sensitivity of the assay allows high throughput format (96-well plate) and evaluation of luciferase activity as soon as after 6 h of incubation, which has potential implication for studies of cytotoxic compounds.
References provided by Crossref.org
Jasmone Is a Ligand-Selective Allosteric Antagonist of Aryl Hydrocarbon Receptor (AhR)
Mixture Effects of Tryptophan Intestinal Microbial Metabolites on Aryl Hydrocarbon Receptor Activity
Targeting the pregnane X receptor using microbial metabolite mimicry
Methylindoles and Methoxyindoles are Agonists and Antagonists of Human Aryl Hydrocarbon Receptor
Enantiospecific effects of ketoconazole on aryl hydrocarbon receptor