Establishment of a Rat Hepatic Cell Line, KanR2-XL8, for a Reporter Gene Assay of Aryl Hydrocarbon Receptor Ligands

To establish a highly sensitive and convenient reporter gene assay for aryl hydrocarbon receptor (AhR) ligands, the chimera plasmid xenobiotic responsible element-luciferase (XRE-Luc) containing an XRE, minimal SV40 promoter, and luciferase reporter gene, was first constructed. The XRE-Luc and the expression vector pRC/CMV containing a neomycin-resistant gene were transfected into a rat hepatic cell line, Kan-R2, and then KanR2-XL8 was selected as a cell clone, which showed the highest response to the induction of luciferase by aryl hydrocarbons, 3-methylcholanthlene (3-MC) and benzo[a]pyrene. Furthermore, AhR-regulated genes, CYP1A1 and CYP1B1, in KanR2-XL8 cells were also activated by 3-MC. In the present study, we established a hepatic cell line, KanR2-XL8, that is useful for screening of AhR ligands with two parameters, the activations of the transfected luciferase gene and the AhR-regulated genes in a host cell.