title: Simian virus 40 large tumor antigen-immortalized normal human liver epithelial cells express hepatocyte characteristics and metabolize chemical carcinogens.
abstract: Normal human liver tissue and cultured human hepatocytes are valuable models to study xenobiotic metabolism and toxicity, but they only have a limited in vitro life-span and are not readily available. This report describes the establishment of replicative cultures of human adult liver epithelial cells in serum-free medium. The longevity of three of these cultures, derived from different donors, was extended by introduction of the simian virus 40 large T antigen gene. Two cell lines, THLE-2 and -3, established with a recombinant simian virus 40 large T antigen virus have undergone > 100 population doublings, are nontumorigenic when injected into athymic nude mice, have near-diploid karyotypes, and do not express alpha-fetoprotein. The cells express cytokeratin 18 and albumin in early passage, whereas higher-passage cells in logarithmic-phase growth also express cytokeratin 19. THLE-2 and -3 cells metabolize benzo[a]pyrene, N-nitrosodimethylamine, and aflatoxin B1 to their ultimate carcinogenic metabolites that adduct DNA, which indicates functional cytochrome P450 pathways. Other enzymes involved in metabolism of chemical carcinogens, such as epoxide hydrolase, NADPH cytochrome P450 reductase, superoxide dismutase, catalase, glutathione S-transferases, and glutathione peroxidase are also retained by THLE cells. Thus, these immortalized human liver cells constitute an in vitro model for pharmacotoxicological studies and for the investigation of etiology and pathogenesis of human hepatocellular carcinoma.
authors.affiliation: Nestec Ltd. Research Center, Lausanne, Switzerland.
authors.forename: A M
authors.initials: AM
authors.lastname: Pfeifer
authors.forename: K E
authors.initials: KE
authors.lastname: Cole
authors.forename: D T
authors.initials: DT
authors.lastname: Smoot
authors.forename: A
authors.initials: A
authors.lastname: Weston
authors.forename: J D
authors.initials: JD
authors.lastname: Groopman
authors.forename: P G
authors.initials: PG
authors.lastname: Shields
authors.forename: J M
authors.initials: JM
authors.lastname: Vignaud
authors.forename: M
authors.initials: M
authors.lastname: Juillerat
authors.forename: M M
authors.initials: MM
authors.lastname: Lipsky
authors.forename: B F
authors.initials: BF
authors.lastname: Trump
mesh_terms: D000328:Adult; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001711:Biotransformation; D015139:Blotting, Southern; D002273:Carcinogens / Q000378:metabolism*; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism; D015183:Restriction Mapping; D012709:Serum Albumin / Q000096:biosynthesis / Q000302:isolation & purification; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection
chemical_list: D000952:Antigens, Polyomavirus Transforming; D002273:Carcinogens; D012333:RNA, Messenger; D012709:Serum Albumin; D012313:RNA; D004247:DNA; D003576:Electron Transport Complex IV