Exocyclic etheno-DNA adducts are formed by the reaction of lipid peroxidation products, such as 4-hydroxy-2-nonenal (HNE) with DNA bases to yield 1,N-6-etheno-2'-deoxyadenosine (epsilon dA), 3,N-4-etheno-2'-deoxycytidine (epsilon dC), and etheno-2'-deoxyguanosine. These adducts act as a driving force for many human malignancies and are elevated in the organs of cancer-prone patients suffering from chronic inflammation and infections. Here, we describe the ultrasensitive and specific techniques for the detection of epsilon dA and epsilon dC in tissue and white blood cell (WBC) DNA. This approach is based on combined immunopurification by monoclonal antibodies and P-32-postlabeling analysis. The detection limit is about five adducts per 10(10) parent nucleotides, requiring 5-10 mu g of DNA. In addition, we describe techniques for immunohistochemical detection of epsilon dA and epsilon dC in tissue biopsies, and the approaches for the analysis of epsilon dA and epsilon dC excreted in urine. The utility of these detection methods for human studies is based on: (1) high sensitivity and specificity, (2) low amounts of DNA required, (3) capability to detect "background" levels of etheno DNA adducts in biopsies, WBC, and urine samples of healthy subjects, and (4) reliable monitoring of the disease-related increase of these substances in patients. The described methods are useful in diagnosis and monitoring of chronic degenerative diseases, including cancer, atherosclerosis, and neurodegenerative disorders.
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第一作者单位：[1]German Canc Res Ctr, Div Toxicol & Canc Risk Factors, Heidelberg, Germany
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推荐引用方式(GB/T 7714)：
Nair Jagadeesan,Nair Urmila J.,Sun Xin,et al.Quantifying Etheno-DNA Adducts in Human Tissues, White Blood Cells, and Urine by Ultrasensitive 32P-Postlabeling and Immunohistochemistry[J].DNA DAMAGE DETECTION IN SITU, EX VIVO, AND IN VIVO: METHODS AND PROTOCOLS.2011,682:189-205.doi:10.1007/978-1-60327-409-8_14.
APA：
Nair, Jagadeesan,Nair, Urmila J.,Sun, Xin,Wang, Ying,Arab, Khelifa&Bartsch, Helmut.(2011).Quantifying Etheno-DNA Adducts in Human Tissues, White Blood Cells, and Urine by Ultrasensitive 32P-Postlabeling and Immunohistochemistry.DNA DAMAGE DETECTION IN SITU, EX VIVO, AND IN VIVO: METHODS AND PROTOCOLS,682,
MLA：
Nair, Jagadeesan,et al."Quantifying Etheno-DNA Adducts in Human Tissues, White Blood Cells, and Urine by Ultrasensitive 32P-Postlabeling and Immunohistochemistry".DNA DAMAGE DETECTION IN SITU, EX VIVO, AND IN VIVO: METHODS AND PROTOCOLS 682.(2011):189-205
