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 共价结合形成 DNA 加合物是化学诱导致癌的第一步。细胞中 DNA 加合物的测量提供了暴露的直接证据以及评估人类健康风险的方法。然而，DNA 加合物形成与人类癌症发展之间的关系仅针对少数致癌物确定。这在一定程度上是因为受影响的组织和器官中加合物的水平较低、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) 结合来明确检测和表征 DNA 加合物。我们的研究通过动物模型证明了毛细管 LC-MS/MS 能够检测和定量 DNA 加合物，其水平与人类接触时遇到的水平相似。我们的工作重点是芳香胺，例如 2-氨基-3-甲基咪唑并[4,5-f]喹啉 (IQ)、4-氨基联苯 (ABP)、2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶 (PhIP) 和 2-氨基-3,8-二甲基咪唑[4,5-f]喹喔啉 (MeIQx)，存在于香烟烟雾中，在烤鱼或牛肉时形成。肺、结肠、乳腺、胰腺和其他组织含有能够将这些致癌物激活成可与 DNA 碱基结合的反应形式的酶。我们建议利用我们成功的动物研究来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。我们将通过将致癌物与模拟 p53 基因序列的合成寡核苷酸反应来进一步检查致癌物的序列识别模式，p53 基因序列已知显示与肺癌相关的特定突变。我们预计该计划生成的数据将有助于确定香烟烟雾与肺癌之间的关系以及 DNA 加合物作为基因毒性癌症指标的价值。