title: Genotype-phenotype correlation between the polymorphic UGT2B17 gene deletion and NNAL glucuronidation activities in human liver microsomes.
abstract: The nicotine-derived tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), is one of the most potent and abundant procarcinogens found in tobacco and tobacco smoke, and glucuronidation of its major metabolite, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), is an important mechanism for NNK detoxification. In cigarette smokers and tobacco chewers, there is a wide variation in the urinary levels of the ratio of NNAL to NNAL glucuronide (NNAL-Gluc). To determine whether genetic variation plays a potential role in this inter-individual variability, NNAL-glucuronidating activities were analysed in a series of human liver microsomal specimens and compared with UGT2B17 deletion genotypes in the same subjects. Assays performed in vitro demonstrated that over-expressed UGT2B17 exhibits high O-glucuronidating activity against NNAL. When stratifying subjects by UGT2B17 genotype, a significant or near-significant decrease in NNAL-O-Gluc formation was observed in liver microsomes from individuals who were either heterozygous [(+/0), P=0.07] or homozygous [(0/0), P=0.016] for the UGT2B17 deletion compared to liver microsomes from individuals with intact UGT2B17 alleles [(+/+)]. There was a significant (P<0.01) association between the level of liver microsomal NNAL-O-glucuronide formation and increasing numbers of the UGT2B17 null alleles in the liver microsomal specimens examined in this study, and a significant decrease in NNAL-O-Gluc formation was observed when comparing liver microsomes from individuals who had at least one UGT2B17 allele deleted [(+/0)+(0/0)] versus microsomes from UGT2B17 (+/+) subjects (P=0.004). When stratifying by the median value of NNAL-O-Gluc formation activity, a significantly (P=0.015) higher number of subjects with liver microsomes having low NNAL-O-Gluc formation activity contained the UGT2B17 null genotype compared to subjects with liver microsomes exhibiting high NNAL-O-Gluc formation activity. When stratifying by UGT2B7/UGT2B17 haplotypes, the association between the level of liver microsomal NNAL-O-glucuronide formation and increasing numbers of the UGT2B17 null allele was at the level of statistical significance for subjects with the UGT2B7 (*1/*2) (P=0.05) or UGT2B7 (*2/*2) (P<0.02) genotypes. These data suggest that the UGT2B17 deletion polymorphism is associated with a reduced rate of NNAL detoxification in vivo and may increase individual susceptibility to tobacco-related cancers.
authors.affiliation: Department of Pharmacology, Penn State University College of Medicine, Hershey, Pennsylvania 17033, USA. plazarus@psu.edu
authors.forename: Philip
authors.initials: P
authors.lastname: Lazarus
authors.forename: Yan
authors.initials: Y
authors.lastname: Zheng
authors.forename: E Aaron
authors.initials: EA
authors.lastname: Runkle
authors.forename: Joshua E
authors.initials: JE
authors.lastname: Muscat
authors.forename: Doris
authors.initials: D
authors.lastname: Wiener
mesh_terms: D001483:Base Sequence; D002273:Carcinogens / Q000378:metabolism; D004247:DNA / Q000235:genetics; D017353:Gene Deletion; D005838:Genotype; D005965:Glucuronates / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000235:genetics* / Q000378:metabolism; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008658:Inactivation, Metabolic; D008862:Microsomes, Liver / Q000378:metabolism*; D015778:Minor Histocompatibility Antigens; D009602:Nitrosamines / Q000378:metabolism*; D010597:Pharmacogenetics; D010641:Phenotype; D011110:Polymorphism, Genetic; D011725:Pyridines / Q000378:metabolism*; D012906:Smoke / Q000032:analysis
chemical_list: C099564:4-((methylnitrosoamino)-1-(3-pyridyl)but-1-yl)beta-omega-glucosiduronic acid; D002273:Carcinogens; D005965:Glucuronates; D015778:Minor Histocompatibility Antigens; D009602:Nitrosamines; D011725:Pyridines; D012906:Smoke; D004247:DNA; C491474:UGT2B7 protein, human; D014453:Glucuronosyltransferase; C500513:UGT2B17 protein, human; C099565:4-(methylnitrosamino)-1-(3-pyridyl)-1-butan-1-ol