Details
| Original language | English |
|---|---|
| Pages (from-to) | 1035-42 |
| Number of pages | 8 |
| Journal | Drug Metabolism and Disposition |
| Volume | 31 |
| Issue number | 8 |
| Publication status | Published - Aug 2003 |
Abstract
Cultures of primary hepatocytes and hepatoma cell line HepG2 are frequently used in in vitro models for human biotransformation studies. In this study, we characterized and compared the capacity of these model systems to indicate the presence of different classes of promutagens. Genotoxic sensitivity, enzyme activity, and gene expression were monitored in response to treatment with food promutagens benzo[a]pyrene, dimethylnitrosamine (DMN), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). DNA damage could be detected reliably with the comet assay in primary human hepatocytes, which were maintained in sandwich culture. All three promutagens caused DNA damage in primary cells, but in HepG2 no genotoxic effects of DMN and PhIP could be detected. We supposed that the lack of specific enzymes accounts for their inability to process these promutagens. Therefore, we quantified the expression of a broad range of genes coding for drug-metabolizing enzymes with real-time reverse transcription-polymerase chain reaction. The genes code for cytochromes p450 and, in addition, for a series of important phase II enzymes. The expression level of these genes in human hepatocytes was similar to those previously reported for human liver samples. On the other hand, expression levels in HepG2 differed significantly from that in human. Activity and expression, especially of phase I enzymes, were demonstrated to be extremely low in HepG2 cells. Up-regulation of specific genes by test substances was similar in both cell types. In conclusion, human hepatocytes are the preferred model for biotransformation in human liver, whereas HepG2 cells may be useful to study regulation of drug-metabolizing enzymes.
Keywords
- 7-Alkoxycoumarin O-Dealkylase/biosynthesis, Benzo(a)pyrene/adverse effects, Carcinoma, Hepatocellular/pathology, Cell Line, Tumor, Comet Assay, Cytochrome P-450 CYP1A1/biosynthesis, Cytoskeleton/ultrastructure, Dimethylnitrosamine/adverse effects, Dose-Response Relationship, Drug, Enzyme Induction/drug effects, Female, Gene Expression/drug effects, Hepatocytes/drug effects, Humans, Imidazoles/pharmacokinetics, Inactivation, Metabolic/genetics, Liver Neoplasms/metabolism, Male, Middle Aged, Mutagens/adverse effects, Reverse Transcriptase Polymerase Chain Reaction
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In: Drug Metabolism and Disposition, Vol. 31, No. 8, 08.2003, p. 1035-42.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Comparison of primary human hepatocytes and hepatoma cell line Hepg2 with regard to their biotransformation properties
AU - Wilkening, Stefan
AU - Stahl, Frank
AU - Bader, Augustinus
PY - 2003/8
Y1 - 2003/8
N2 - Cultures of primary hepatocytes and hepatoma cell line HepG2 are frequently used in in vitro models for human biotransformation studies. In this study, we characterized and compared the capacity of these model systems to indicate the presence of different classes of promutagens. Genotoxic sensitivity, enzyme activity, and gene expression were monitored in response to treatment with food promutagens benzo[a]pyrene, dimethylnitrosamine (DMN), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). DNA damage could be detected reliably with the comet assay in primary human hepatocytes, which were maintained in sandwich culture. All three promutagens caused DNA damage in primary cells, but in HepG2 no genotoxic effects of DMN and PhIP could be detected. We supposed that the lack of specific enzymes accounts for their inability to process these promutagens. Therefore, we quantified the expression of a broad range of genes coding for drug-metabolizing enzymes with real-time reverse transcription-polymerase chain reaction. The genes code for cytochromes p450 and, in addition, for a series of important phase II enzymes. The expression level of these genes in human hepatocytes was similar to those previously reported for human liver samples. On the other hand, expression levels in HepG2 differed significantly from that in human. Activity and expression, especially of phase I enzymes, were demonstrated to be extremely low in HepG2 cells. Up-regulation of specific genes by test substances was similar in both cell types. In conclusion, human hepatocytes are the preferred model for biotransformation in human liver, whereas HepG2 cells may be useful to study regulation of drug-metabolizing enzymes.
AB - Cultures of primary hepatocytes and hepatoma cell line HepG2 are frequently used in in vitro models for human biotransformation studies. In this study, we characterized and compared the capacity of these model systems to indicate the presence of different classes of promutagens. Genotoxic sensitivity, enzyme activity, and gene expression were monitored in response to treatment with food promutagens benzo[a]pyrene, dimethylnitrosamine (DMN), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). DNA damage could be detected reliably with the comet assay in primary human hepatocytes, which were maintained in sandwich culture. All three promutagens caused DNA damage in primary cells, but in HepG2 no genotoxic effects of DMN and PhIP could be detected. We supposed that the lack of specific enzymes accounts for their inability to process these promutagens. Therefore, we quantified the expression of a broad range of genes coding for drug-metabolizing enzymes with real-time reverse transcription-polymerase chain reaction. The genes code for cytochromes p450 and, in addition, for a series of important phase II enzymes. The expression level of these genes in human hepatocytes was similar to those previously reported for human liver samples. On the other hand, expression levels in HepG2 differed significantly from that in human. Activity and expression, especially of phase I enzymes, were demonstrated to be extremely low in HepG2 cells. Up-regulation of specific genes by test substances was similar in both cell types. In conclusion, human hepatocytes are the preferred model for biotransformation in human liver, whereas HepG2 cells may be useful to study regulation of drug-metabolizing enzymes.
KW - 7-Alkoxycoumarin O-Dealkylase/biosynthesis
KW - Benzo(a)pyrene/adverse effects
KW - Carcinoma, Hepatocellular/pathology
KW - Cell Line, Tumor
KW - Comet Assay
KW - Cytochrome P-450 CYP1A1/biosynthesis
KW - Cytoskeleton/ultrastructure
KW - Dimethylnitrosamine/adverse effects
KW - Dose-Response Relationship, Drug
KW - Enzyme Induction/drug effects
KW - Female
KW - Gene Expression/drug effects
KW - Hepatocytes/drug effects
KW - Humans
KW - Imidazoles/pharmacokinetics
KW - Inactivation, Metabolic/genetics
KW - Liver Neoplasms/metabolism
KW - Male
KW - Middle Aged
KW - Mutagens/adverse effects
KW - Reverse Transcriptase Polymerase Chain Reaction
U2 - 10.1124/dmd.31.8.1035
DO - 10.1124/dmd.31.8.1035
M3 - Article
C2 - 12867492
VL - 31
SP - 1035
EP - 1042
JO - Drug Metabolism and Disposition
JF - Drug Metabolism and Disposition
SN - 0090-9556
IS - 8
ER -