A consecutive three alanine residue insertion mutant of human CAR: a novel CAR ligand screening system in HepG2 cells.

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Kanno Y, Inouye Y

A consecutive three alanine residue insertion mutant of human CAR: a novel CAR ligand screening system in HepG2 cells.

J Toxicol Sci. 2010 Aug;35(4):515-25.

PubMed ID

20686338 [ View in PubMed

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Abstract

The expression of the CAR gene is lost in most cultured cell lines, and exogenously expressed CAR accumulates spontaneously in the nuclear compartment, where CAR is constitutively active. Therefore, the assessment of CAR-dependent transcriptional activation has to be evaluated using either animal models or primary cultured hepatocytes. Furthermore, due to the species-specific response of CAR to individual compounds, the results obtained with animal models are not directly applicable to human cases. We established a novel screening system for hCAR ligands (including agonists and inverse agonists) by expressing GAL4/DBD-fused hCAR/LBD, in which three consecutive alanine residues were inserted between helix 11 and helix 12 of LBD, and a commercially obtained plasmid consisting of the firefly luciferase gene downstream of the GAL4 responsive element in a cultured cell line. Androstenol (Andro) and clotrimazole (CLT), which are both inverse agonists of hCAR, were classified as an antagonist and weak agonist, respectively, in our novel assay system. Among DDT and its metabolites DDE and DDD, only DDT worked as an agonist of the hCAR mutant, although all of them were enrolled as agonists of SXR. Using this system, the screening for hCAR ligands can be conducted without sacrificing animals.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Clotrimazole	Hydroxycarboxylic acid receptor 2	Protein	Humans	Unknown	Partial agonist	Details

Pharmaco-transcriptomics

Drug	Drug Groups	Gene	Gene ID	Change	Interaction	Chromosome
Phenobarbital	Approved Investigational	CYP2B6	1555	upregulated	Phenobarbital results in increased expression of CYP2B6 mRNA	19q13.2
Phenobarbital	Approved Investigational	CYP3A4	1576	upregulated	Phenobarbital results in increased expression of CYP3A4 mRNA	7q22.1