Graduation Year

2004

Document Type

Dissertation

Degree

Ph.D.

Degree Granting Department

Public Health

Major Professor

Ira Richards, Ph.D.

Committee Member

Raymond D. Harbison, Ph.D.

Committee Member

Jong Y. Park, Ph.D.

Committee Member

Philip Roets, Sc.D.

Keywords

Polymorphisms, Benzo[a]pyrene, Colon carcinogenesis, Pharmacogenetics, Orolaryngeal Cancer

Abstract

Exposure to polycyclic aromatic hydrocarbons (PAHs) such as benzo(a)pyrene are important risk factors for cancer. Three UDP-glucuronosyltransferases, UGT1A9, UGT1A10, and UGT2B7, have been shown to play an important role in the phase II metabolism of carcinogenic metabolites of BaP. Because UGT1A9 and UGT2B7 are well-expressed in digestive tract tissues including liver and colon, it is possible that genetic variations in either enzyme may play an important role in colon cancer risk. However, UGT1A10 is extrahepatic and is expressed in the oral cavity and the larynx; therefore, genetic variations in this enzyme may play an important role in risk for orolaryngeal cancer. This study examined UGT1A9-, UGT1A10-, and UGT2B7-specific sequences for polymorphisms that play a role in cancer susceptibility. For the UGT1A9 gene, two missense polymorphisms at codons 167 (Val>Ala) and 183 (Cys>Gly) were identified. A previously-reported missense polymorphism was identified for the UGT2B7 gene. To assess the potential role of UGT1A10 variants as a risk factor for orolaryngeal cancer, PCR-RFLP was used to identify UGT1A10 genotypes in DNA specimens isolated from 115 African American newly-diagnosed orolaryngeal cancer cases and 115 non-cancer controls individually matched by age and race. A significantly decreased risk for orolaryngeal cancer was observed for subjects possessing one or more UGT1A10139Lys alleles as determined by crude analysis or after logistic regression analysis adjusting for age, sex, smoking and alcohol consumption. These results strongly suggest that the UGT1A10139Lys polymorphism may play an important protective role in risk for orolaryngeal cancer. To determine whether the change in amino acid sequence at codon 183 results in aberrant UGT1A9 enzyme activity, functional characterization of the wild-type- and variant-encoded UGT1A9 isoforms was performed in vitro. Cell homogenates were prepared from UGT1A9-transfected HK293 cells and glucuronidation assays were performed against various carcinogens/carcinogen metabolites. A significant (p<0.001) 3- to 4-fold decrease in enzyme activity, determined by HPLC analysis, was observed for the UGT1A9183Gly variant as compared to its wild-type counterpart for all substrates analyzed. These results demonstrate that the UGT1A9 (Cys183Gly) polymorphism significantly alters UGT1A9 function and could potentially play an important role as risk modifier for digestive tract cancers.

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