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J.Health Sci., 50(5), 530-536, 2004
Establishment of a Rat Hepatic Cell Line,
KanR2-XL8, for a Reporter Gene Assay of Aryl Hydrocarbon Receptor Ligands
Masashi Sekimoto, Miho Iwamoto, Shoji Miyajima, Kiyomitsu Nemoto, and Masakuni Degawa*
Department of Molecular Toxicology and COE Program in the 21st Century, School of Pharmaceutical Sciences, University of
Shizuoka, 52-1 Yada, Shizuoka 422-8526, Japan
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.
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