Establishment of a Human Hepatoma Cell Line HepG2-A10 for a Reporter Gene Assay of Arylhydrocarbon Receptor Activators

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  • Sekimoto Masashi
    Department of Molecular Toxicology and COE Program in the 21st Century, School of Pharmaceutical Sciences, University of Shizuoka
  • Kawamagari Hiroto
    Department of Molecular Toxicology and COE Program in the 21st Century, School of Pharmaceutical Sciences, University of Shizuoka
  • Nakatani Saki
    Department of Molecular Toxicology and COE Program in the 21st Century, School of Pharmaceutical Sciences, University of Shizuoka
  • Nemoto Kiyomitsu
    Department of Molecular Toxicology and COE Program in the 21st Century, School of Pharmaceutical Sciences, University of Shizuoka
  • Degawa Masakuni
    Department of Molecular Toxicology and COE Program in the 21st Century, School of Pharmaceutical Sciences, University of Shizuoka

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To establish a human hepatic cell line for a convenient reporter gene assay of arylhydrocarbon receptor (AhR) activators, the chimera plasmid containing xenobiotic responsible element (XRE), minimal SV40 promoter, and luciferase reporter gene and the expression vector pRC/CMV containing a neomycin-resistant gene were co-transfected into a human hepatoma cell line, HepG2. Then, antibiotic (G418)-resistant HepG2 cells were selected and cloned. A cell clone, HepG2-A10, showed the highest responsibility to 3-methylcholanthrene (MC)-mediated induction of luciferase among the clones obtained. Expression levels of luciferase activity in HepG2-A10 cells were increased in a dose- and time-dependent manner by treatment with either MC, an AhR-ligand type activator, or omeprazole (OME), a non-AhR ligand type activator. In addition, expression levels of cytochrome P4501A subfamily genes (CYP1A1 and CYP1A2) were also increased in a dose- and time-dependent manner by treatment with MC. The present findings demonstrate that a newly established human HepG2-A10 is a useful cell line for a convenient reporter gene assay of AhR activators.<br>

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