title: Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells.
abstract: Most human hepatocyte cell lines lack a substantial set of liver-specific functions, especially major cytochrome P450 (P450)-related enzyme activities, making them unrepresentative of in vivo hepatocytes. We have used the HepaRG cells, derived from a human hepatocellular carcinoma, which exhibit a high differentiation pattern after 2 weeks at confluency to determine whether they could mimic human hepatocytes for drug metabolism and toxicity studies. We show that when passaged at low density, these cells reversed to an undifferentiated morphology, actively divided, and, after having reached confluency, formed typical hepatocyte-like colonies surrounded by biliary epithelial-like cells. By contrast, when seeded at high density, hepatocyte-like clusters retained their typical differentiated morphology. Transcripts of various nuclear receptors (aryl hydrocarbon receptor, pregnane X receptor, constitutive androstane receptor, peroxisome proliferator-activated receptor alpha), P450s (CYP1A2, 2C9, 2D6, 2E1, 3A4), phase 2 enzymes (UGT1A1, GSTA1, GSTA4, GSTM1), and other liver-specific functions were estimated by reverse transcriptase-quantitative polymerase chain reaction and were found to be expressed, for most of them, at comparable levels in both confluent differentiated and high-density differentiated HepaRG cells and in cultured primary human hepatocytes. For several transcripts, the levels were strongly increased in the presence of 2% dimethyl sulfoxide. Measurement of basal activities of several P450s and their response to prototypical inducers as well as analysis of metabolic profiles and cytotoxicity of several compounds confirmed the functional resemblance of HepaRG cells to primary cultured human hepatocytes. In conclusion, HepaRG cells constitute the first human hepatoma cell line expressing high levels of the major P450s involved in xenobiotic metabolism and represent a reliable surrogate to human hepatocytes for drug metabolism and toxicity studies.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM) U620, Université de Rennes 1, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Aninat
authors.forename: Amélie
authors.initials: A
authors.lastname: Piton
authors.forename: Denise
authors.initials: D
authors.lastname: Glaise
authors.forename: Typhen
authors.initials: T
authors.lastname: Le Charpentier
authors.forename: Sophie
authors.initials: S
authors.lastname: Langouët
authors.forename: Fabrice
authors.initials: F
authors.lastname: Morel
authors.forename: Christiane
authors.initials: C
authors.lastname: Guguen-Guillouzo
authors.forename: André
authors.initials: A
authors.lastname: Guillouzo
mesh_terms: D000082:Acetaminophen / Q000494:pharmacology; D016604:Aflatoxin B1 / Q000494:pharmacology; D000638:Amiodarone / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002746:Chlorpromazine / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D014453:Glucuronosyltransferase / Q000235:genetics*; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D007527:Isoenzymes / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015262:Xenobiotics / Q000494:pharmacology
chemical_list: D007527:Isoenzymes; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D015262:Xenobiotics; D000082:Acetaminophen; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1; D014453:Glucuronosyltransferase; D000638:Amiodarone; D002746:Chlorpromazine; D004121:Dimethyl Sulfoxide