ROLE OF 02 RADICALS AND IRON IN ASBESTO-INDUCED CANCER

02 自由基和铁在石棉诱发的癌症中的作用

• 批准号: 2701315
• 负责人: ANN E AUST
• 金额: $ 13.92万
• 依托单位: UTAH STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2701315
• 关键词: DNA damage; animal tissue; asbestos; biological signal transduction; catalyst; chemical carcinogenesis; clone cells; cocarcinogen; cytotoxicity; environment related neoplasm /cancer; free radical oxygen; free radical scavengers; gel mobility shift assay; human tissue; hydrogen peroxide; iron; nitric oxide; nitric oxide synthase; northern blottings; protein kinase; tissue /cell culture

DESCRIPTION: (Applicant's Abstract) The long-term objective of this research is to investigate the role of reactive oxygen species, now to include nitric oxide (NO), and iron in asbestos-induced cancer. The hypothesis for the proposed research is that asbestos activates cell signalling pathways in target cells which lead to induction of NO synthesis and that NO and iron associated with asbestos participate in chemical reactions leading to DNA damage and mutations. The Specific Aims for the proposed research are: 1) to determine whether asbestos is activating cell signalling pathways which are responsible for the induction of an inducible form of nitric oxide synthase (iNOS) in human cells; 2) to compare the amount of mRNA for iNOS with respect to time after crocidolite treatment of human cells with cytokine or chrysotile treatments; 3) to determine how iron and NO participate to cause oxidative damage to DNA in crocidolite-treated cells; and 4) to determine the effect of iron and/or NO on the mutagenicity of crocidolite in the gpt+ transgenic hprt V79 cell lines, G10 and G12. The effect of crocidolite-treatment of human lung epithelial cells (A549) and mesothelial cells (JMN and Met-5A) on activation of NF-kB by protein kinases will be determined by cotreatment with various protein kinase inhibitors followed by electrophoretic gel mobility shift assays of nuclear extracts. The time course of NF-kB activation, lipid peroxidation, and DNA oxidation will be compared to elucidate which signalling pathway(s) may be involved. The significant differences between the induction of mRNA for iNOS by crocidolite, chrysotile and cytokines will be determined by Northern analysis of RNA. Immunostaining techniques, using antibodies specific for nitrotyrosine, will be used to determine whether peroxynitrite is being formed in these cells. DNA repair enzymes for 8-hydroxydeoxyguanosine and uracil glycosylase will be assayed in treated cells to determine whether NO has inhibited their repair function. Mutagenesis studies in transgenic V79 cell lines sensitive to reactive oxygen species will be done to determine whether both iron and NO are required for mutations induce by crocidolite.

描述：（申请人的摘要）本发明的长期目标是： 研究是调查活性氧的作用，现在， 包括一氧化氮（NO）和铁。的 这项研究的假设是石棉激活细胞 导致NO诱导的靶细胞中的信号通路 合成和NO和与石棉相关铁参与 化学反应导致DNA损伤和突变。具体 本研究的目的是：1）确定石棉 激活了细胞信号通路， 诱导型一氧化氮合酶（iNOS）在人中诱导 细胞; 2）比较iNOS mRNA的量随时间的变化 在青石棉用细胞因子或温石棉处理人细胞后 治疗; 3）确定铁和NO如何参与导致 青石棉处理的细胞中对DNA的氧化损伤;以及4） 确定铁和/或NO对致突变性的影响 gpt+转基因hprt V79细胞系G10和G12中的青石棉。 青石棉对人肺上皮细胞的作用 （A549）和间皮细胞（JMN和Met-5A）对NF-kB活化的影响。 蛋白激酶将通过与各种蛋白质共处理来测定 激酶抑制剂，随后进行电泳凝胶迁移率变动测定 核提取物。NF-kB激活的时间过程，脂质 过氧化和DNA氧化将进行比较，以阐明 可能涉及信号通路。的显著差异 青石棉、温石棉和 细胞因子将通过RNA的北方分析来确定。 使用硝基酪氨酸特异性抗体的免疫染色技术， 将被用来确定是否过氧亚硝酸盐正在形成这些 细胞8-羟基脱氧鸟苷和尿嘧啶的DNA修复酶 将在处理的细胞中测定糖基化酶以确定NO是否具有 抑制了它们的修复功能转基因V79的诱变研究 细胞系敏感的活性氧物种将进行，以确定 是否铁和NO都是诱导突变所必需的 青石棉