DNA Adducts of Aromatic Amines: Analysis by HPLC-MS

芳香胺 DNA 加合物：HPLC-MS 分析

• 批准号: 7022934
• 负责人: Paul Vouros
• 金额: $ 23万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-18 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7022934
• 关键词: DNA damage; adduct; amines; biomarker; breast neoplasms; cancer risk; capillary electrophoresis; carbopolycyclic compound; cell line; chemical carcinogenesis; clinical research; electrospray ionization mass spectrometry; high performance liquid chromatography; human tissue; liquid chromatography; lung neoplasms; mass spectrometry; mutagen testing; nuclear magnetic resonance spectroscopy; oligonucleotides; smoking

DESCRIPTION (provided by applicant): The covalent binding of genotoxins to DNA to form DNA adducts is considered by many to be the first step in chemically induced carcinogenesis. Measurement of DNA adducts in cells provides direct evidence for exposure and a means for assessing human health risk. However, the relationship between DNA adduct formation and cancer development in humans has only been established for a few carcinogens. This is, in part, because of the low levels at which adducts occur in affected tissues and organs, the limited amounts of DNA available, and the difficulties associated with the analysis. In order to address these analytical problems, we have been exploring the use of capillary separation methods (liquid chromatography (LC) and capillary electrophoresis (CE)) coupled to mass spectrometry (MS) for the unequivocal detection and characterization of DNA adducts. Using animal models, our research has demonstrated the capability of capillary LC-MS/MS for the detection and quantification of DNA adducts at levels similar to those encountered in human exposure. Our work has focused on aromatic amines such as 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 4-aminobiphenyl (ABP), 2-amino-1- methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), found in cigarette smoke and formed during the grilling of fish or beef. Lung, colon, breast, pancreas and other tissues contain enzymes capable of activating these carcinogens into reactive forms that can bind to DNA bases. We propose to capitalize on our successful animal studies to examine lung and other human tissues with emphasis on the comparison of DNA adduct content between smokers and non-smokers. We will further examine the sequence recognition patterns of carcinogens by reacting them with synthetic oligonucleotides that mimic p53 gene sequences known to display specific mutations associated with lung cancer. We expect that the data generated in this program will help define the relationship between cigarette smoke and lung cancer and the value of DNA adducts as indicators for genotoxic cancer.

描述（由申请人提供）：许多人认为基因毒素与DNA形成DNA加合物的共价结合是化学诱导的癌变的第一步。细胞中DNA加合物的测量为暴露和评估人类健康风险的手段提供了直接证据。但是，仅针对少数致癌物建立了DNA加合物形成与癌症发展之间的关系。这部分是因为在受影响的组织和器官中发生加合物，可用的DNA量有限以及与分析相关的困难。为了解决这些分析问题，我们一直在探索使用毛细管分离方法（液相色谱（LC）和毛细管电泳（CE））与质谱（MS）的使用，以实现不等式的DNA加合物的表征。使用动物模型，我们的研究证明了毛细血管LC-MS/MS的能力，可以在类似于人类暴露的水平上检测和定量DNA加合物的水平。我们的工作集中在芳香胺上，例如2-氨基-3-甲基咪唑唑[4,5-F]喹啉（IQ），4-氨基苯基苯基（ABP），2-氨基-1-甲基-1-甲基-6-苯基咪唑唑[4,5-B]吡啶（PHIP）和2-氨基-3-氨基-3-二氨基二氨基二氨基[4,5-B] （meiqx），在香烟中发现，在鱼或牛肉烧烤期间形成。肺，结肠，乳腺癌，胰腺和其他组织含有能够将这些致癌物激活成反应性形式的酶，这些形式可以与DNA碱基结合。我们建议利用我们成功的动物研究，以检查肺和其他人体组织，重点是比较吸烟者和非吸烟者之间的DNA加合物含量。我们将进一步检查致癌物的序列识别模式，使它们与合成寡核苷酸反应，以模仿已知的p53基因序列，这些p53基因序列已知显示与肺癌相关的特定突变。我们预计该程序中产生的数据将有助于定义香烟烟与肺癌之间的关系，以及DNA加合物的价值作为遗传毒性癌的指标。