title: Host determinants of DNA alkylation and DNA repair activity in human colorectal tissue: O(6)-methylguanine levels are associated with GSTT1 genotype and O(6)-alkylguanine-DNA alkyltransferase activity with CYP2D6 genotype.
abstract: There is increasing evidence that alkylating agent exposure may increase large bowel cancer risk and factors which either alter such exposure or its effects may modify risk. Hence, in a cross-sectional study of 78 patients with colorectal disease, we have examined whether (i) metabolic genotypes (GSTT1, GSTM1, CYP2D6, CYP2E1) are associated with O(6)-methyldeoxyguanosine (O(6)-MedG) levels, O(6)-alkylguanine-DNA alkyltransferase (ATase) activity or K-ras mutations, and (ii) there was an association between ATase activity and O(6)-MedG levels. Patients with colon tumours and who were homozygous GSTT1(*)2 genotype carriers were more likely than patients who expressed GSTT1 to have their DNA alkylated (83 versus 32%, P=0.03) and to have higher O(6)-MedG levels (0.178+/-0.374 versus 0.016+/-0.023 micromol O(6)-MedG/mol dG, P=0.04) in normal, but not tumour, DNA. No such association was observed between the GSTT1 genotype and the frequency of DNA alkylation or O(6)-MedG levels in patients with benign colon disease or rectal tumours. Patients with colon tumours or benign colon disease who were CYP2D6-poor metabolisers had higher ATase activity in normal tissue than patients who were CYP2D6 extensive metabolisers or CYP2D6 heterozygotes. Patients with the CYP2E1 Dra cd genotype were less likely to have a K-ras mutation: of 55 patients with the wild-type CYP2E1 genotype (dd), 23 had K-ras mutations, whereas none of the 7 individuals with cd genotype had a K-ras mutation (P=0.04). No other associations were observed between GSTT1, GSTM1, CYP2D6 and CYP2E1 Pst genotypes and adduct levels, ATase activity or mutational status. O(6)-MedG levels were not associated with ATase activity in either normal or tumour tissue. However, in 15 patients for whom both normal and tumour DNA contained detectable O(6)-MedG levels, there was a strong positive association between the normal DNA/tumour DNA adduct ratio and the normal tissue/tumour tissue ATase ratio (r(2)=0.66, P=0.001). These results indicate that host factors can affect levels both of the biologically effective dose arising from methylating agent exposure and of a susceptibility factor, the DNA repair phenotype.
authors.affiliation: Cancer Research Campaign Carcinogenesis Group, Paterson Institute for Cancer Research, Manchester M20 9BX, UK. andy.povey@man.ac.uk
authors.forename: A C
authors.initials: AC
authors.lastname: Povey
authors.forename: C N
authors.initials: CN
authors.lastname: Hall
authors.forename: A F
authors.initials: AF
authors.lastname: Badawi
authors.forename: D P
authors.initials: DP
authors.lastname: Cooper
authors.forename: M J
authors.initials: MJ
authors.lastname: Guppy
authors.forename: P E
authors.initials: PE
authors.lastname: Jackson
authors.forename: P J
authors.initials: PJ
authors.lastname: O'Connor
authors.forename: G P
authors.initials: GP
authors.lastname: Margison
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000368:Aged; D000478:Alkylation; D015179:Colorectal Neoplasms / Q000201:enzymology* / Q000235:genetics; D003430:Cross-Sectional Studies; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D004260:DNA Repair; D004273:DNA, Neoplasm / Q000378:metabolism*; D005260:Female; D020022:Genetic Predisposition to Disease; D005838:Genotype; D005982:Glutathione Transferase / Q000235:genetics* / Q000378:metabolism; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000378:metabolism*; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism
chemical_list: D004273:DNA, Neoplasm; D006147:Guanine; C008449:O-(6)-methylguanine; D019389:Cytochrome P-450 CYP2D6; D019853:O(6)-Methylguanine-DNA Methyltransferase; C413545:glutathione S-transferase T1; D005982:Glutathione Transferase; D000251:Adenosine Triphosphatases; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)