Analysis of acrolein-derived 1,N2-propanodeoxyguanosine adducts in human leukocyte DNA from smokers and nonsmokers.

Cigarette smoking is a major source of human exposure to acrolein, a widespread environmental pollutant and toxicant that is also formed endogenously through metabolism of amino acids and polyamines and lipid peroxidation. Acrolein reacts with DNA, producing two pairs of regioisomeric 1,N(2)-propanodeoxyguanosine adducts: (6R/S)-3-(2'-deoxyribos-1'-yl)-5,6,7,8-tetrahydro-6-hydroxypyrimido[1,2-a]purine-10(3H)one (α-OH-Acr-dGuo) and (8R/S)-3-(2'-deoxyribos-1'-yl)-5,6,7,8-tetrahydro-8-hydroxypyrimido[1,2-a]purine-10(3H)one (γ-OH-Acr-dGuo). Previous studies indicate that these adducts might be involved in producing mutations in the p53 tumor suppressor gene, as observed in lung tumors in smokers, but there are only limited published data comparing acrolein-DNA adducts in smokers and nonsmokers. In this study, we developed a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method to analyze Acr-dGuo adducts in human leukocyte DNA. The potential for artifactual formation was found in two steps of the assay: DNA isolation and DNA hydrolysis. This was eliminated by employing a Ficoll-Hypaque double density gradient to obtain leukocytes free of erythrocyte contamination and by adding glutathione to scavenge acrolein present in H(2)O. The accuracy and precision of the method were confirmed. Acr-dGuo adducts were analyzed in leukocyte DNA from 25 smokers and 25 nonsmokers. γ-OH-Acr-dGuo was the predominant isomer in all samples, while α-OH-Acr-dGuo was detected in only three subjects. There was no significant difference between the total Acr-dGuo levels in smokers (7.4 ± 3.4 adducts/10(9) nucleotides) and nonsmokers (9.8 ± 5.5 adducts/10(9) nucleotides). Although our study is limited in size, these results, together with the results of previous analyses of acrolein-derived mercapturic acids in the urine of smokers and nonsmokers, suggest that glutathione conjugation effectively removes acrolein from external exposures such as cigarette smoking, protecting leukocyte DNA from damage.

[1]  C. Rizzo,et al.  Formation of deoxyguanosine cross-links from calf thymus DNA treated with acrolein and 4-hydroxy-2-nonenal. , 2010, Chemical research in toxicology.

[2]  Jian-Min Yuan,et al.  Applying tobacco carcinogen and toxicant biomarkers in product regulation and cancer prevention. , 2010, Chemical research in toxicology.

[3]  I. Minko,et al.  Chemistry and Biology of DNA Containing 1,N2-Deoxyguanosine Adducts of the α,β-Unsaturated Aldehydes Acrolein, Crotonaldehyde, and 4-Hydroxynonenal , 2009, Chemical research in toxicology.

[4]  D. Hatsukami,et al.  Effects of smoking cessation on eight urinary tobacco carcinogen and toxicant biomarkers. , 2009, Chemical research in toxicology.

[5]  L. Marnett,et al.  Formation of DNA−Protein Cross-Links Between γ-Hydroxypropanodeoxyguanosine and EcoRI , 2008, Chemical research in toxicology.

[6]  A. Cheema,et al.  Detection of the acrolein-derived cyclic DNA adduct by a quantitative 32P-postlabeling/solid-phase extraction/HPLC method: blocking its artifact formation with glutathione. , 2008, Analytical biochemistry.

[7]  J. F. Stevens,et al.  Acrolein: sources, metabolism, and biomolecular interactions relevant to human health and disease. , 2008, Molecular nutrition & food research.

[8]  D. Hatsukami,et al.  Quantitation of acrolein-derived (3-hydroxypropyl)mercapturic acid in human urine by liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry: effects of cigarette smoking. , 2007, Chemical research in toxicology.

[9]  S. Hecht,et al.  Detection and quantitation of acrolein-derived 1,N2-propanodeoxyguanosine adducts in human lung by liquid chromatography-electrospray ionization-tandem mass spectrometry. , 2007, Chemical research in toxicology.

[10]  J. Michael.  Textbook of Medical Physiology , 2005 .

[11]  R. Carchman,et al.  Chemical composition, cytotoxicity and mutagenicity of smoke from US commercial and reference cigarettes smoked under two sets of machine smoking conditions. , 2004, Toxicology.

[12]  R. Hayes.  Dry cleaning, some chlorinated solvents and other industrial chemicals IARC monographs on the evaluation of carcinogenic risks to humans. volume 63 , 1996, Cancer Causes & Control.

[13]  S. Orrenius,et al.  Pharmacokinetics and mechanism of action of detoxifying low-molecular-weight thiols , 2004, Journal of Cancer Research and Clinical Oncology.

[14]  I. Minko,et al.  Comparative Evaluation of the Bioreactivity and Mutagenic Spectra of Acrolein-Derived α-HOPdG and γ-HOPdG Regioisomeric Deoxyguanosine Adducts , 2003 .

[15]  R. Lloyd,et al.  1,N 2-Deoxyguanosine Adducts of Acrolein, Crotonaldehyde, and trans-4-Hydroxynonenal Cross-link to Peptides via Schiff Base Linkage* , 2003, The Journal of Biological Chemistry.

[16]  I. Minko,et al.  Comparative evaluation of the bioreactivity and mutagenic spectra of acrolein-derived alpha-HOPdG and gamma-HOPdG regioisomeric deoxyguanosine adducts. , 2003, Chemical research in toxicology.

[17]  F. Johnson,et al.  Mutagenesis by acrolein-derived propanodeoxyguanosine adducts in human cells. , 2002, Biochemistry.

[18]  N. Tretyakova,et al.  Tobacco smoke carcinogens, DNA damage and p53 mutations in smoking-associated cancers , 2002, Oncogene.

[19]  William Rom,et al.  Preferential DNA damage and poor repair determine ras gene mutational hotspot in human cancer. , 2002, Journal of the National Cancer Institute.

[20]  J. Goodisman,et al.  Kinetic analysis of the reactions of 4-hydroperoxycyclophosphamide and acrolein with glutathione, mesna, and WR-1065. , 2002, Drug metabolism and disposition: the biological fate of chemicals.

[21]  F. Chung,et al.  Formation of cyclic deoxyguanosine adducts from ω-3 and ω-6 polyunsaturated fatty acids under oxidative conditions , 2002 .

[22]  F. Chung,et al.  Formation of cyclic deoxyguanosine adducts from omega-3 and omega-6 polyunsaturated fatty acids under oxidative conditions. , 2002, Chemical research in toxicology.

[23]  F. Johnson,et al.  NMR Characterization of a DNA Duplex Containing the Major Acrolein-derived Deoxyguanosine Adduct γ-OH-1,-N 2-Propano-2′-deoxyguanosine* , 2001, The Journal of Biological Chemistry.

[24]  F. Chung,et al.  1,N2-propanodeoxyguanosine adducts: potential new biomarkers of smoking-induced DNA damage in human oral tissue. , 1998, Cancer research.

[25]  R. Talaat,et al.  Metabolism and distribution of [2,3-14C]acrolein in Sprague-Dawley rats. II. Identification of urinary and fecal metabolites. , 1998, Toxicological sciences : an official journal of the Society of Toxicology.

[26]  F. Chung,et al.  Effect of glutathione depletion on exocyclic adduct levels in the liver DNA of F344 rats. , 1997, Chemical research in toxicology.

[27]  F. Chung,et al.  Detection of 1, N2-propanodeoxyguanosine adducts as potential endogenous DNA lesions in rodent and human tissues. , 1996, Cancer research.

[28]  F. Chung,et al.  Detection of exocyclic 1,N2-propanodeoxyguanosine adducts as common DNA lesions in rodents and humans. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  S. Hecht,et al.  Formation of cyclic 1,N2-propanodeoxyguanosine adducts in DNA upon reaction with acrolein or crotonaldehyde. , 1984, Cancer research.

[30]  A. Prasad,et al.  Zinc in plasma, neutrophils, lymphocytes, and erythrocytes as determined by flameless atomic absorption spectrophotometry. , 1982, Clinical chemistry.