Quantitation by Liquid Chromatography-Nanoelectrospray Ionization-High-Resolution Tandem Mass Spectrometry of Multiple DNA Adducts Related to Cigarette Smoking in Oral Cells in the Shanghai Cohort Study.
Quantitation by Liquid Chromatography-Nanoelectrospray Ionization-High-Resolution Tandem Mass Spectrometry of Multiple DNA Adducts Related to Cigarette Smoking in Oral Cells in the Shanghai Cohort Study.
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DOI：
10.1021/acs.chemrestox.2c00393
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发表时间：
2023-02-20
影响因子：
4.1
通讯作者：
Hecht, Stephen S.
中科院分区：
文献类型：
--
作者：
Cheng, Guang;Guo, Jiehong;Wang, Renwei;Yuan, Jian-Min;Balbo, Silvia;Hecht, Stephen S.
我们开发了液相色谱-纳米电喷雾电离-高分辨率串联质谱（LC-NSI-HRMS/MS）方法，用于同时定量分析与吸烟相关的5种口腔细胞DNA加合物：（8R/S）-3-（2 '-脱氧核糖-1'-基）-5，6，7，8-四氢-8-羟基嘧啶并[1，2-a]嘌呤-10（3 H）-酮（α-OH-Acr-dGuo，1）由丙烯醛，（6S，8S和6R，8R）-3-（2 '-脱氧核糖-1'-基）-5，6，7，8-四氢-8-羟基-6-甲基嘧啶并[1，2-a]嘌呤-10（3 H）-酮[（6S，8 S）-OH-Cro-dGuo，2]和[（6 R，8 R）-OH-Cro-dGuo，3]、丙烯腈、氯乙烯、脂质过氧化和炎症引起的1，N6-乙烯基-dAdo（4）和氧化损伤引起的8-oxo-dGuo（5）。在谷胱甘肽存在下分离口腔细胞DNA以防止伪影形成。获得了清晰的LC-NSI-HRMS/MS色谱图，允许使用适当标记的内标物定量各加合物。对该方法的准确度和精密度进行了验证，加合物1-4的含量测定定量限为5 fmol/µmol dGuo，加合物5的含量测定定量限为20 fmol/µmol。该测定应用于从上海队列研究中收集的80个口腔细胞样本; 40个来自当前吸烟者，40个来自从不吸烟者。吸烟者和非吸烟者之间的所有加合物水平存在显着差异。在吸烟者和非吸烟者中，8-oxo-dGuo（5）的水平至少是其他加合物的3,000倍，吸烟者与非吸烟者之间这种加合物的量之间的差异虽然显著（P = 0.013），但没有吸烟者与非吸烟者之间其他DNA加合物的差异那么大（P值均小于0.001）。本研究提供了一种新的LC-NSI-HRMS/MS方法，用于定量由接触α，β-不饱和醛丙烯醛和巴豆醛、炎症和氧化损伤引起的各种DNA加合物，这些都与致癌作用有关。我们预期这种检测方法在正在进行的与吸烟相关的肺癌和口腔癌的分子流行病学研究中的应用。
We developed a liquid chromatography-nanoelectrospray ionization-high resolution tandem mass spectrometry (LC-NSI-HRMS/MS) method for simultaneous quantitative analysis of 5 oral cell DNA adducts associated with cigarette smoking: (8R/S)-3-(2’-deoxyribos-1’-yl)-5,6,7,8-tetrahydro-8-hydroxypyrimido[1,2-a]purine-10(3H)-one (ɣ-OH-Acr-dGuo, 1) from acrolein, (6S,8S and 6R,8R)-3-(2’-deoxyribos-1’-yl)-5,6,7,8-tetrahydro-8-hydroxy-6-methylpyrimido[1,2-a]purine-10(3H)-one [(6S,8S)ɣ-OH-Cro-dGuo, 2] and [(6R,8R)ɣ-OH-Cro-dGuo, 3] from crotonaldehyde, 1,N6-etheno-dAdo (4) from acrylonitrile, vinyl chloride, lipid peroxidation and inflammation, and 8-oxo-dGuo (5) from oxidative damage. Oral cell DNA was isolated in the presence of glutathione to prevent artifact formation. Clear LC-NSI-HRMS/MS chromatograms were obtained allowing quantitation of each adduct using the appropriately labelled internal standards. The accuracy and precision of the method were validated and the assay limit of quantitation was 5 fmol/µmol dGuo for adducts 1-4 and 20 fmol/µmol for adduct 5. The assay was applied to 80 buccal cell samples selected from those collected in the Shanghai Cohort Study; 40 from current smokers and 40 from never smokers. Significant differences were found in all adduct levels between smokers and non-smokers. Levels of 8-oxo-dGuo (5) were at least 3,000 times greater than those of the other adducts in both smokers and non-smokers, and the differences between amounts of this adduct in smokers versus non-smokers, while significant (P = 0.013), was not as great as the differences of the other DNA adducts between smokers and non-smokers (P values all less than 0.001). No significant relationship of adduct levels to risk of lung cancer incidence was found. This study provides new LC-NSI-HRMS/MS methodology for the quantitation of diverse DNA adducts resulting from exposure to the α,β-unsaturated aldehydes acrolein and crotonaldehye, inflammation, and oxidative damage which are all associated with carcinogenesis. We anticipate application of this assay in ongoing studies of the molecular epidemiology of cancers of the lung and oral cavity related to cigarette smoking.
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影响因子：
4.1
作者：
Hecht, Stephen S.
通讯作者：
Hecht, Stephen S.
影响因子：
4.7
作者：
Cheng, Guang;Guo, Jiehong;Hecht, Stephen S.
通讯作者：
Hecht, Stephen S.
DOI：
10.1158/1940-6207.capr-09-0192
发表时间：
2010-03
期刊：
Cancer prevention research (Philadelphia, Pa.)
影响因子：
--
作者：
Boyle JO;Gümüs ZH;Kacker A;Choksi VL;Bocker JM;Zhou XK;Yantiss RK;Hughes DB;Du B;Judson BL;Subbaramaiah K;Dannenberg AJ
通讯作者：
Dannenberg AJ
影响因子：
4.3
作者：
Li, Yupeng;Hecht, Stephen S.
通讯作者：
Hecht, Stephen S.
影响因子：
3.3
作者：
Bhutani, Manisha;Pathak, Ashutosh Kumar;Mao, Li
通讯作者：
Mao, Li