RRC ID 38514
著者 Hosomi H, Fukami T, Iwamura A, Nakajima M, Yokoi T.
タイトル Development of a highly sensitive cytotoxicity assay system for CYP3A4-mediated metabolic activation.
ジャーナル Drug Metab Dispos
Abstract Drug-induced hepatotoxicity, which is a rare but serious adverse reaction to a large number of pharmaceutical drugs, is sometimes associated with reactive metabolites produced by drug-metabolizing enzymes. In the present study, we constructed a cell-based system to evaluate the cytotoxicity of reactive metabolites produced by CYP3A4 using human hepatoma cells infected with an adenovirus vector expressing human CYP3A4 (AdCYP3A4). When seven hepatoma cell lines (HepG2, Hep3B, HLE, HLF, Huh6, Huh7, and Fa2N4 cells) were infected with AdCYP3A4, HepG2 cells showed the highest CYP3A4 protein expression and testosterone 6β-hydroxylase activity (670 pmol · min(-1) · mg(-1)). With the use of AdCYP3A4-infected HepG2 cells, the cytotoxicities of 23 drugs were evaluated by the 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt assay, and the cell viability when treated with 11 drugs (amiodarone, desipramine, felbamate, isoniazid, labetalol, leflunomide, nefazodone, nitrofurantoin, tacrine, terbinafine, and tolcapone) was significantly decreased. Moreover, the transfection of siRNA for nuclear factor erythroid 2-related factor 2 (Nrf2) to decrease the cellular expression level of Nrf2 exacerbated the cytotoxicity of some drugs (troglitazone, flutamide, acetaminophen, clozapine, terbinafine, and desipramine), suggesting that the genes regulated by Nrf2 are associated with the detoxification of the cytotoxicities mediated by CYP3A4. We constructed a highly sensitive cell-based system to detect the drug-induced cytotoxicity mediated by CYP3A4. This system would be beneficial in preclinical screening in drug development and increase our understanding of the drug-induced cytotoxicity associated with CYP3A4.
巻・号 39(8)
ページ 1388-95
公開日 2011-8-1
DOI 10.1124/dmd.110.037077
PII dmd.110.037077
PMID 21540358
MeSH Biological Assay / methods* Cell Line, Tumor Cell Survival / drug effects Chemical and Drug Induced Liver Injury / etiology Cytochrome P-450 CYP3A / metabolism* Drug Design Drug Evaluation, Preclinical / methods* Drug-Related Side Effects and Adverse Reactions* Enzyme Activation Humans NF-E2-Related Factor 2 / metabolism Sensitivity and Specificity
IF 3.231
引用数 38
WOS 分野 PHARMACOLOGY & PHARMACY
リソース情報
ヒト・動物細胞