Metabolic activation and carcinogen-DNA adduct detection in human larynx.
Putative carcinogen-DNA adducts in human larynx tissues (n = 25) from smoker and non/ex-smoker patients were examined by 32P-postlabeling and compared with the metabolic activation capacity of larynx microsomes and cytosols from the same tissues. Hydrophobic DNA adducts were evident only in smokers, and chromatographic profiles of the adducts were similar using either the butanol extraction or nuclease P1 enhancement method, which suggested that the adducts may be derived from polycyclic aromatic hydrocarbons but not aromatic amines. Immunoblots of larynx microsomes using anti-cytochrome P450 1A1/1A2, 2C, 3A4, 2E1, and 2A6 antibodies showed intensities ranging from 1-10% of that typically observed with human liver microsomes. Enzymatic assays of larynx microsomes showed appreciable activity for benzo(a)pyrene hydroxylation (P450 1A1 and 2C) but not for 4-aminobiphenyl N-oxidation (P450 1A2), which indicated that the observed immunoreactivity was for P450 1A1; this represents the highest level of this P450 yet detected in human extrahepatic tissues. Accordingly, total DNA adduct levels in the larynx correlated strongly with levels of P450 2C, 1A1, and 3A4 but not with P450 2E1 or 2A6. Larynx cytosols also showed appreciable aromatic amine N-acetyl-transferase activity for p-aminobenzoic acid (NAT-1) but not for sulfamethazine (NAT-2); however, NAT-1 activity was not correlated with total DNA adducts, which is again consistent with the lack of aromatic amine-DNA adducts detected by 32P-postlabeling. Thus, these results suggest that the DNA adducts detected in human larynx are largely derived from metabolic activation of polycyclic aromatic hydrocarbons in cigarette smoke by P450 2C, 3A4, and/or 1A1.
Degawa
M
M
Office of Research (HFT-100), National Center for Toxicological Research, Jefferson, Arkansas 72079.
Stern
S J
SJ
Martin
M V
MV
Guengerich
F P
FP
Fu
P P
PP
Ilett
K F
KF
Kaderlik
R K
RK
Kadlubar
F F
FF
D000123:Acetyltransferases / Q000032:analysis; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D004273:DNA, Neoplasm / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D007822:Laryngeal Neoplasms / Q000737:chemistry*; D008861:Microsomes / Q000737:chemistry; D009092:Mucous Membrane / Q000378:metabolism; D011083:Polycyclic Compounds / Q000032:analysis* / Q000378:metabolism; D012907:Smoking / Q000378:metabolism
D004273:DNA, Neoplasm; D011083:Polycyclic Compounds; D003577:Cytochrome P-450 Enzyme System; D000123:Acetyltransferases