Detection of the acrolein-derived cyclic DNA adduct by a quantitative 32P-postlabeling/solid-phase extraction/HPLC method: Blocking its artifact formation with glutathione

Emami, Armaghan; Dyba, Marcin; Cheema, Amrita K.; Pan, Jason; Nath, Raghu G.; Chung, Fung‐Lung

doi:10.1016/j.ab.2007.10.029

Cited by 24 publications

(33 citation statements)

References 17 publications

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“…19,24 The Chung group first reported the formation of Acr-dGuo as an artifact during the analysis, and added GSH to block acrolein reaction with DNA. 24 We later adopted the same scavenger to inhibit artifacts for the analysis of Acr-dGuo in human leukocytes. 19 The measured Acr-dGuo levels were 20-50% lower when GSH was used during the sample preparation.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, we developed a more sensitive assay using liquid chromatography-nanoelectrospray ionization-high resolution tandem mass spectrometry (LC-NSI-HRMS/MS). Artifactual formation of Acr-dGuo during sample preparation has been observed recently, 24 and no precautionary steps to block artifact formation were included in our earlier study. 18 In this study, we have modified our previous method to lower the levels of artifacts formed during sample preparation and minimize their interference in quantitation.…”

Section: Introductionmentioning

confidence: 99%

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Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Yang

Balbo

Villalta

et al. 2019

Chem. Res. Toxicol.

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Acrolein, the simplest α, β-unsaturated aldehyde, is present in relatively large quantities in cigarette smoke and several studies have raised the possibility of it being a major etiological agent for smoking related lung cancer. Acrolein reacts directly with DNA to form primarily Acr-dGuo adducts, which serve as important biomarkers for the assessment of exposure to acrolein and its potential role in smoking related lung cancer. In this study, we developed an ultra-sensitive and low-artifact method using liquid chromatography-nanoelectrospray ionization-high resolution tandem mass spectrometry (LC-NSI-HRMS/MS) to quantitate Acr-dGuo adducts in normal lung tissue DNA obtained at surgery from lung cancer patients who never smoked and from those who continued smoking until surgery, as confirmed by urinary total cotinine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). This provides a direct comparison of Acr-dGuo levels in human lung tissue as a result of cigarette smoking versus other etiological causes. There was no significant difference between the total Acr-dGuo levels in smokers (28.5 ± 14.9 adducts/10 9 nucleotides) and non-smokers (25.0 ± 10.7 adducts/10 9 nucleotides), suggesting rapid removal of acrolein by glutathione conjugation and other detoxification mechanisms. Our results do not support the hypothesis that acrolein is a major etiological agent for cigarette smoking related DNA damage.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Yang

Balbo

Villalta

et al. 2019

Chem. Res. Toxicol.

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“…We examined the effects of cigarette smoking on the levels of Acr-dGuo in leukocytes from smokers and non-smokers. In developing this assay, we took into account the recent observations by the Chung group that Acr-dGuo adducts can form readily as artifacts during analysis of DNA (18). …”

Section: Introductionmentioning

confidence: 99%

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Leukocyte DNA from Smokers and Nonsmokers

Zhang

Balbo

Wang

et al. 2010

Chem. Res. Toxicol.

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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-AcrdGuo). 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 non-smokers. 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. 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 non-smokers. γ-OH-Acr-dGuo was the predominant isomer in all samples, while α-OH-Acr-dGuo was detected in only 3 subjects. There was no significant difference between the total Acr-dGuo levels in smokers (7.4 ± 3.4 adducts/10 9 nucleotides) and non-smokers (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 non-smokers, suggest that glutathione conjugation effectively removes acrolein from external exposures such as cigarette smoking, protecting leukocyte DNA from damage.

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“…The HFD+TE diet was the same high fat diet above, supplemented with 2% TE (Tokyo, Japan). The 2% TE was chosen based on previous reported bioassays (23). Diet was stored at 4 o C and changed weekly.…”

Section: Methodsmentioning

confidence: 99%

Prevention of Lipid Peroxidation–derived Cyclic DNA Adduct and Mutation in High-Fat Diet–induced Hepatocarcinogenesis by Theaphenon E

Coia

Hou

et al. 2018

Cancer Prevention Research

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Obesity is associated with cancer risk and its link with liver cancer is particularly strong. Obesity causes non-alcoholic fatty liver disease (NAFLD) that could progress to hepatocellular carcinoma (HCC). Chronic inflammation likely plays a key role. We carried out a bioassay in the high-fat diet (HFD)-fed C57BL/6J mice to provide insight into the mechanisms of obesity-related HCC by studying γ-OHPdG, a mutagenic DNA adduct derived from lipid peroxidation. In an 80-week bioassay, mice received a low-fat diet (LFD), high-fat diet (HFD), and HFD with 2% Theaphenon E (TE) (HFD+TE). HFD mice developed a 42% incidence of HCC and LFD mice a 16%. Remarkably, TE, a standardized green tea extract formulation, completely blocked HCC in HFD mice with a 0% incidence. γ-OHPdG measured in the hepatic DNA of mice fed HFD and HFD+TE showed its levels increased during the early stages of NAFLD in HFD mice and the increases were significantly suppressed by TE, correlating with the tumor data. Whole-exome sequencing showed an increased mutation load in the liver tumors of HFD mice with G>A and G>T as the predominant mutations, consistent with the report that γ-OHPdG induces G>A and G>T. Furthermore, the mutation loads were significantly reduced in HFD+TE mice, particularly G>T, the most common mutation in human HCC. These results demonstrate in a relevant model of obesity-induced HCC that γ-OHPdG formation during fatty liver disease may be an initiating event for accumulated mutations that leads to HCC and this process can be effectively inhibited by TE. .

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Detection of the acrolein-derived cyclic DNA adduct by a quantitative 32P-postlabeling/solid-phase extraction/HPLC method: Blocking its artifact formation with glutathione

Cited by 24 publications

References 17 publications

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Leukocyte DNA from Smokers and Nonsmokers

Prevention of Lipid Peroxidation–derived Cyclic DNA Adduct and Mutation in High-Fat Diet–induced Hepatocarcinogenesis by Theaphenon E

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Detection of the acrolein-derived cyclic DNA adduct by a quantitative 32P-postlabeling/solid-phase extraction/HPLC method: Blocking its artifact formation with glutathione

Cited by 24 publications

References 17 publications

Analysis of Acrolein-Derived 1,N2-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Analysis of Acrolein-Derived 1,N2-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Analysis of Acrolein-Derived 1,N2-Propanodeoxyguanosine Adducts in Human Leukocyte DNA from Smokers and Nonsmokers

Prevention of Lipid Peroxidation–derived Cyclic DNA Adduct and Mutation in High-Fat Diet–induced Hepatocarcinogenesis by Theaphenon E

Contact Info

Product

Resources

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Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Lung DNA from Smokers and Nonsmokers

Analysis of Acrolein-Derived 1,N²-Propanodeoxyguanosine Adducts in Human Leukocyte DNA from Smokers and Nonsmokers