Polymorphic drug metabolism: studies with recombinant Chinese hamster cells and analyses in human populations.
Most promutagens and procarcinogens exert their genotoxicity after undergoing metabolic activation. Metabolism of chemicals is an important factor in limiting the extent of the action of a chemical. In this study, we established cell lines which carried cDNAs coding for human CYP1A2 and N-acetyltransferase (NAT); the latter functions as O-acetyltransferase for N-hydroxyarylamines formed by CYP1A2. A cell line which expresses CYP1A2 together with P450 reductase activated aflatoxin B1, but not heterocyclic amines. A cell line which carries CYP1A2 and polymorphic NAT (NAT2) in addition to P450 reductase efficiently activated IQ and some other heterocyclic amines. However, a cell line which carries CYP1A2 and monomorphic NAT (NAT1) had only low activity toward the same heterocyclic amines. In order to determine the presence of and frequency of genetic polymorphisms of CYP1A2 and NAT2 in humans, we performed in caffeine phenotyping test on 205 Japanese volunteers. Analyses of metabolic ratios of urinary metabolites showed a bimodal distribution, indicating that about 86% and 91% of Japanese were extensive metabolizers (EM) of CYP1A2 and NAT2, respectively. The genotype NAT2 determined by the PCR-RFLP method agreed completely with the phenotype. To determine the mechanism of the differences in CYP1A2 activity, genomic DNA from peripheral lymphocytes of poor metabolizers (PM) and EM was subjected to DNA sequencing. No differences in nucleotide sequence were observed between PMs and EMs in the exons, exon-intron junctions and 5'-flanking region of the CYP1A2 gene.(ABSTRACT TRUNCATED AT 250 WORDS)
Yokoi
T
T
Division of Drug Metabolism, Faculty of Pharmaceutical Sciences, Hokkaido University, Japan.
Sawada
M
M
Kamataki
T
T
D000818:Animals; D001483:Base Sequence; D001711:Biotransformation; D002273:Carcinogens / Q000378:metabolism* / Q000633:toxicity; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D019388:Cytochrome P-450 CYP1A2; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D005091:Exons; D006801:Humans; D008969:Molecular Sequence Data; D009153:Mutagens / Q000378:metabolism* / Q000633:toxicity; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D011355:Prodrugs / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D014162:Transfection
D002273:Carcinogens; D009153:Mutagens; D011355:Prodrugs; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019388:Cytochrome P-450 CYP1A2