RRC ID 67510
著者 Iwado S, Abe S, Oshimura M, Kazuki Y, Nakajima Y.
タイトル Bioluminescence Measurement of Time-Dependent Dynamic Changes of CYP-Mediated Cytotoxicity in CYP-Expressing Luminescent HepG2 Cells.
ジャーナル Int J Mol Sci
Abstract We sought to develop a cell-based cytotoxicity assay using human hepatocytes, which reflect the effects of drug-metabolizing enzymes on cytotoxicity. In this study, we generated luminescent human hepatoblastoma HepG2 cells using the mouse artificial chromosome vector, in which click beetle luciferase alone or luciferase and major drug-metabolizing enzymes (CYP2C9, CYP2C19, CYP2D6, and CYP3A4) are expressed, and monitored the time-dependent changes of CYP-mediated cytotoxicity expression by bioluminescence measurement. Real-time bioluminescence measurement revealed that compared with CYP-non-expressing cells, the luminescence intensity of CYP-expressing cells rapidly decreased when the cells were treated with low concentrations of aflatoxin B1 or primaquine, which exhibits cytotoxicity in the presence of CYP3A4 or CYP2D6, respectively. Using kinetics data obtained by the real-time bioluminescence measurement, we estimated the time-dependent changes of 50% inhibitory concentration (IC50) values in the aflatoxin B1- and primaquine-treated cell lines. The first IC50 value was detected much earlier and at a lower concentration in primaquine-treated CYP-expressing HepG2 cells than in primaquine-treated CYP-non-expressing cells, and the decrease of IC50 values was much faster in the former than the latter. Thus, we successfully monitored time- and concentration-dependent dynamic changes of CYP-mediated cytotoxicity expression in CYP-expressing luminescent HepG2 cells by means of real-time bioluminescence measurement.
巻・号 22(6)
公開日 2021-3-11
DOI 10.3390/ijms22062843
PII ijms22062843
PMID 33799598
PMC PMC7999318
MeSH Aflatoxin B1 / toxicity* Animals Cell Line, Tumor Cytochrome P-450 CYP2C19 / genetics Cytochrome P-450 CYP2C19 / metabolism Cytochrome P-450 CYP2C9 / genetics Cytochrome P-450 CYP2C9 / metabolism Cytochrome P-450 CYP2D6 / genetics Cytochrome P-450 CYP2D6 / metabolism Cytochrome P-450 CYP3A / genetics Cytochrome P-450 CYP3A / metabolism Founder Effect* Gene Expression Regulation Genes, Reporter Genetic Vectors / chemistry Genetic Vectors / metabolism Hep G2 Cells Humans Inhibitory Concentration 50 Luciferases / genetics Luciferases / metabolism Luminescence Luminescent Measurements / methods* Mice Primaquine / toxicity* Time-Lapse Imaging / methods* Xenobiotics / toxicity*
IF 4.556
リソース情報
ヒト・動物細胞 Hep G2(RCB1886)