OXIDIZED DNA BASES AS BIOMARKERS OF TUMOR PROGRESSION

氧化 DNA 碱基作为肿瘤进展的生物标志物

• 批准号: 7977079
• 负责人: Paul Thomas Henderson
• 金额: $ 6.83万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-04 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7977079
• 关键词: 8-hydroxyguanosine; Animals; Biological Assay; Biological Markers; Breast Cancer Cell; Cancer Etiology; Cells; Chemoprevention; Clinical; Code; Computer Retrieval of Information on Scientific Projects Database; DNA; DNA Adducts; DNA Damage; DNA Markers; Detection; Diagnostic; Environmental Risk Factor; Epidemiologic Studies; Forensic Medicine; Foundations; Fractionation; Free Radicals; Funding; Future; Grant; Human; Institution; Lead; Link; Malignant Neoplasms; Malignant neoplasm of lung; Measurement; Measures; Metabolism; Methods; Mutation; Operative Surgical Procedures; Oxygen; Proteins; Rattus; Research; Research Personnel; Resource Development; Resources; Sampling; Skin Cancer; Smoking; Source; Sun Exposure; Techniques; United States National Institutes of Health; accelerator mass spectrometry; base; carcinogenesis; malignant breast neoplasm; new technology; oxidation; oxidative DNA damage; polypeptide; theories; tumor progression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. An early theory of carcinogenesis proposes that DNA is damaged by environmental factors, which lead to changes in the DNA code. These changes are called mutations. Once enough mutations accumulate, cells can no longer function properly, and cancer arises. The facts that smoking causes lung cancer, and that sunlight exposure causes skin cancer support this theory. However, such a direct environmental link is not evident for breast cancer in humans. A more recent theory suggests that the metabolism of oxygen produces free radicals that cause carcinogenic DNA damage. In order to validate the latter theory, it is necessary to measure the products of free radical damage in cells. Damage to cells caused by reactive oxygen is called oxidation. In some breast cancers, increased levels of oxidative DNA damage have been associated with tumor progression. However, the most commonly used marker of DNA oxidation, called 8-oxoG, is known to be chemically unstable, and is difficult to measure accurately. Importantly, 8-oxoG is itself easily oxidized to form several secondary oxidation products. We propose to develop new technology that will allow detection of the products in rat and human breast cancer cells. Such an assay can be used as a foundation for making diagnostics for use in future human clinical and epidemiological studies. An additional benefit of these biomarkers will be their eventual application to chemoprevention in animals and humans. AMS is a desirable technique due to its sensitivity and robustness as a method for high-throughput measurement. In comparison, less quantitative MS methods such as FT-MS lack the precision of AMS for routine operations, especially at the expected level of a few DNA adducts per cell. The combination of these two techniques will allow for the fractionation and detection of protein polypeptides at or below attomole levels, which is necessary for small forensic samples. This project is complementary aims to those of Project 3 of the AMS RR grant, particularly with regard to the use of AMS to further understand cancer etiology using DNA-based markers.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 早期的致癌理论认为，DNA 会受到环境因素的破坏，从而导致 DNA 密码发生变化。这些变化称为突变。 一旦积累了足够多的突变，细胞就无法正常发挥作用，癌症就会出现。 吸烟导致肺癌和阳光照射导致皮肤癌的事实支持了这一理论。 然而，这种直接的环境联系对于人类乳腺癌并不明显。最近的理论表明，氧的代谢会产生自由基，从而导致致癌的 DNA 损伤。 为了验证后一种理论，有必要测量细胞中自由基损伤的产物。 活性氧对细胞造成的损害称为氧化。 在一些乳腺癌中，氧化 DNA 损伤水平的增加与肿瘤进展有关。 然而，最常用的 DNA 氧化标记物（称为 8-oxoG）化学性质不稳定，难以准确测量。 重要的是，8-oxoG 本身很容易被氧化形成多种二次氧化产物。 我们建议开发新技术，以检测大鼠和人类乳腺癌细胞中的产品。 这种测定可以作为未来人类临床和流行病学研究中使用的诊断的基础。 这些生物标志物的另一个好处是它们最终应用于动物和人类的化学预防。 AMS 因其作为高通量测量方法的灵敏度和稳健性而成为一种理想的技术。 相比之下，定量较少的 MS 方法（例如 FT-MS）在常规操作中缺乏 AMS 的精度，尤其是在每个细胞中存在少量 DNA 加合物的预期水平时。 这两种技术的结合将允许在阿托摩尔水平或以下的蛋白质多肽的分级和检测，这对于小法医样品是必要的。 该项目是对 AMS RR 赠款项目 3 目标的补充，特别是在使用 AMS 通过基于 DNA 的标记进一步了解癌症病因方面。