title: Upgrading cytochrome P450 activity in HepG2 cells co-transfected with adenoviral vectors for drug hepatotoxicity assessment.
abstract: In a number of adverse drug reactions leading to hepatotoxicity, drug metabolism is thought to be involved by the generation of reactive metabolites from non-toxic drugs. The use of hepatoma cell lines, such as HepG2 cell line, for the evaluation of drug-induced hepatotoxicity is hampered by their low cytochrome P450 expression which makes impossible the study of the toxicity produced by bioactivable compounds. Genetically manipulated cells constitute promising tools for hepatotoxicity applications. HepG2 cells were simultaneously transfected with recombinant adenoviruses encoding CYP1A2, CYP2C9 and CYP3A4 to confer them drug-metabolic competence. Upgraded cells (Adv-HepG2) were highly able to metabolize the toxin studied in contrast to the reduced metabolic capacity of HepG2 cells. Aflatoxin B1-induced hepatotoxicity was studied as a proof of concept in metabolically competent and non-competent HepG2 cells by using high content screening technology. Significant differences in mitochondrial membrane potential, intracellular calcium concentration, nuclear morphology and cell viability after treatment with aflatoxin B1 were observed in Adv-HepG2 when compared to HepG2 cells. Rotenone (non bioactivable) and citrate (non hepatotoxic) were analysed as negative controls. This cell model showed to be a suitable hepatic model to test hepatotoxicity of bioactivable drugs and constitutes a valuable alternative for hepatotoxicity testing.
authors.affiliation: Unidad de Hepatología Experimental, Centro de Investigación, Hospital La Fe, Valencia, Spain.
authors.forename: Laia
authors.initials: L
authors.lastname: Tolosa
authors.forename: M Teresa
authors.initials: MT
authors.lastname: Donato
authors.forename: Gabriela
authors.initials: G
authors.lastname: Pérez-Cataldo
authors.forename: José Vicente
authors.initials: JV
authors.lastname: Castell
authors.forename: M José
authors.initials: MJ
authors.lastname: Gómez-Lechón
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D016604:Aflatoxin B1 / Q000378:metabolism / Q000633:toxicity*; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000473:pathology; D019343:Citric Acid / Q000008:administration & dosage / Q000378:metabolism; D019388:Cytochrome P-450 CYP1A2 / Q000235:genetics; D065729:Cytochrome P-450 CYP2C9; D051544:Cytochrome P-450 CYP3A / Q000235:genetics; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008954:Models, Biological; D012402:Rotenone / Q000008:administration & dosage / Q000378:metabolism; D014162:Transfection
chemical_list: D012402:Rotenone; D019343:Citric Acid; D016604:Aflatoxin B1; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D002118:Calcium