Detection of DNA adducts of acetaldehyde in peripheral white blood cells of alcohol abusers

Abstract

The effect of alcohol drinking on the formation of DNA adducts of acetaldehyde, the primary oxidative metabolite of ethanol, was investigated in humans. DNA was isolated from granulocytes and lymphocytes from 24 alcoholic patients and 12 control subjects. DNA adduct levels were measured by 32P-postlabelling using reversed-phase HPLC with on-line detection of radioactivity. A large interindividual variation in adduct levels was observed. The average adduct levels in granulocyte and lymphocyte DNA from alcoholic patients were 3.4 +/- 3.8 and 2.1 +/- 0.8 adducts/10(7) nucleotides (n = 24), respectively. These levels were 13- and 7-fold higher than the corresponding levels in control subjects (P<0.001). The average adduct level in granulocyte DNA from alcoholic patients was 60% higher than in lymphocyte DNA (P<0.01). Our results, in conjunction with the genotoxicity of acetaldehyde, thus suggest the formation of DNA adducts of acetaldehyde as a plausible mechanism explaining the involvement of alcohol drinking in carcinogenesis.

Quantitation of an Acetaldehyde Adduct in Human Leukocyte DNA and the Effect of Smoking Cessation

“…We have used a similar method to quantify N 2 -ethylidene-dGuo in human liver DNA (5). A previous study quantified N 2 -ethyl-dGuo in granulocytes and lymphocytes of alcoholic patients and controls using 32 Ppostlabelling (9). Levels in alcoholic patients were 2-3 adducts per 10 7 nucleotides, but the adduct was not detected in control subjects.…”

Risk factors and mechanisms of hepatocarcinogenesis with special emphasis on alcohol and oxidative stress

“…Binding to DNA and the formation of stable adducts represents one mechanism by which acetaldehyde could trigger the occurrence of replication errors and/or mutations in oncogenes or tumor suppressor genes. The occurrence of stable DNA adducts has been shown in different organs of alcohol-fed rodents and in leukocytes of alcoholics (Fang and Vaca, 1997). It has been shown that the major stable DNA adduct N 2 -ethyl desoxyguanosine (N2-Et-dG) indeed serves as a substrate of eukaryotic DNA polymerase (Matsuda et al, 1999).…”

Structure-Function Relationships in Miscoding by Sulfolobus solfataricus DNA Polymerase Dpo4: GUANINE N2,N2-DIMETHYL SUBSTITUTION PRODUCES INACTIVE AND MISCODING POLYMERASE COMPLEXES

“…Pyridyloxobutyl-derived N 2 -alkyl G adducts have been detected in rats after chronic treatment with NЈ-nitrosonornicotine (31). N 2 -EtG was detected in granulocyte and lymphocyte DNA and urine of alcoholic patients (32,33), and misincorporation opposite N 2 -EtG by Escherichia coli DNA polymerase I/Klenow fragment (exonuclease Ϫ ) has been reported (22). Crotonaldehyde-and acetaldehyde-derived 1,N 2 -propanodeoxyguanosine adducts have also been characterized in human tissue DNA (25).…”