Constitutive and inducible expression of CYP enzymes in immortal hepatocytes derived from SV40 transgenic mice.
1. The expression of liver-specific transcription factors and cytochrome P450 (CYP) enzymes have been studied in three new hepatocyte-like cell lines derived from SV Delta 202 transgenic mice: AMH-Delta 202 (adult mouse hepatocytes), TAMH-Delta 202 (tumour-derived adult mouse hepatocytes) and NMH-Delta 202 (newborn mouse hepatocytes). 2. mRNA levels of liver-enriched transcription factors such as D-element binding protein (DBP), liver-enriched transcription activating protein (LAP) and the hepatic nuclear factors (HNF) 1, 2 and 3 in all Delta 202 transgenic hepatocyte lines were similar to those in the wild-type liver and in primary mouse hepatocytes. 3. Analysis of basal CYP activities and testosterone metabolism revealed that Delta 202 cells showed higher similarities to mouse hepatocytes than Hepa 1c1c7 hepatoma cells. All three Delta 202 cell lines exhibited substantial active CYP1A1/2, CYP2A4/5 and CYP3A11 activities and lower levels of CYP2B, CYP2C and CYP2E1 activities. 4. The Delta 202 cells also responded to model inducers. 3-Methylcholanthrene induced CYP1A1/2 (7-ethoxyresorufin O-deethylation); phenobarbital induced CYP2B (7-benzoxyresorufin O-debenzylation), CYP2A4/5 (testosterone 7alpha -hydroxylation) and CYP3A11 (testosterone 6beta -hydroxylation); and rifampicin and dexamethasone induced CYP3A11 activities in the three Delta 202 cell lines, whereas only AMH-Delta 202 cells reproduced to a limited extent the response of CYP2E1 to ethanol observed in hepatocytes. 5. The results suggest that generation of hepatocyte lines from transgenic animals constitutes a successful approach to obtain in vitro models alternative to primary hepatocytes for drug metabolism and CYP inducibility studies.
Unidad de Hepatología Experimental, Centro de Investigación, Hospital Univesitario La Fe, Avda. Campanar 21, E-46009 Valencia, Spain.
M T
MT
Donato
R
R
Klocke
J V
JV
Castell
K
K
Stenzel
D
D
Paul
M J
MJ
Gómez-Lechón
D000818:Animals; D002460:Cell Line; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology*; D006900:Hydroxylation; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008748:Methylcholanthrene / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D010634:Phenobarbital / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012293:Rifampin / Q000494:pharmacology; D013739:Testosterone / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
D012333:RNA, Messenger; D014157:Transcription Factors; D000431:Ethanol; D013739:Testosterone; D008748:Methylcholanthrene; D003907:Dexamethasone; D003577:Cytochrome P-450 Enzyme System; D012293:Rifampin; D010634:Phenobarbital