Cooke Marcus S, Rozalski Rafal, Dove Rosamund, Gackowski Daniel, Siomek Agnieszka, Evans Mark D, Olinski Ryszard
Department of Cancer Studies and Molecular Medicine, Leicester Royal Infirmary, University Hospitals of Leicester NHS Trust, Leicester, LE2 7LX, UK.
Biol Chem. 2006 Apr;387(4):393-400. doi: 10.1515/BC.2006.053.
Measurement of the products of oxidatively damaged DNA in urine is a frequently used means by which oxidative stress may be assessed non-invasively. We believe that urinary DNA lesions, in addition to being biomarkers of oxidative stress, can potentially provide more specific information, for example, a reflection of repair activity. We used high-performance liquid chromatography prepurification, with gas chromatography-mass spectrometry (LC-GC-MS) and ELISA to the analysis of a number of oxidative [e.g., 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), 8-oxo-7,8-dihydro-guanine, 5-(hydroxymethyl)uracil], non-oxidative (cyclobutane thymine dimers) and oligomeric DNA products in urine. We analysed spot urine samples from 20 healthy subjects, and 20 age- and sex-matched cancer patients. Mononuclear cell DNA 8-oxodG levels were assessed by LC-EC. The data support our proposal that urinary DNA lesion products are predominantly derived from DNA repair. Furthermore, analysis of DNA and urinary 8-oxodG in cancer patients and controls suggested reduced repair activity towards this lesion marker in these patients.
测量尿液中氧化损伤DNA的产物是一种常用的手段，通过它可以非侵入性地评估氧化应激。我们认为，尿液中的DNA损伤除了是氧化应激的生物标志物外，还可能提供更具体的信息，例如，反映修复活性。我们使用高效液相色谱预纯化，结合气相色谱-质谱联用（LC-GC-MS）和酶联免疫吸附测定（ELISA）来分析尿液中的多种氧化产物[如8-氧代-7,8-二氢-2'-脱氧鸟苷（8-氧代dG）、8-氧代-7,8-二氢鸟嘌呤、5-（羟甲基）尿嘧啶]、非氧化产物（环丁烷胸腺嘧啶二聚体）和寡聚DNA产物。我们分析了20名健康受试者以及20名年龄和性别匹配的癌症患者的随机尿样。通过液相色谱-电化学检测法评估单核细胞DNA的8-氧代dG水平。数据支持我们的观点，即尿液中的DNA损伤产物主要来自DNA修复。此外，对癌症患者和对照组的DNA及尿液8-氧代dG的分析表明，这些患者对该损伤标志物的修复活性降低。
