Reactive oxygen species in Ni(II) carcinogenesis

Ni(II) 致癌过程中的活性氧

• 批准号: 8035981
• 负责人: Xianglin Shi
• 金额: $ 32.31万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8035981
• 关键词: Animal Experiments; Animals; Antioxidants; Biological Assay; Breathing; Carcinogenicity Tests; Carcinogens; Cells; Chemicals; Confocal Microscopy; Development; Dominant-Negative Mutation; Down-Regulation; Early Diagnosis; Electron Spin Resonance Spectroscopy; Embryo; Enzymes; Epidemiology; Epithelial Cells; Fibroblasts; Gene Expression Regulation; Generations; Goals; Human; Hydrogen Peroxide; Hydroxyl Radical; I Kappa B-Alpha; In Vitro; Inbred BALB C Mice; Intervention; Investigation; Link; Luciferases; Malignant Neoplasms; Manganese Superoxide Dismutase; Measures; Mediating; Mediator of activation protein; Methods; Mitochondria; Molecular; Mus; NADPH Oxidase; Nature; Nickel; Nuclear; Nude Mice; Plasmids; Play; Prevention strategy; Protein Overexpression; Reaction; Reactive Oxygen Species; Reporter; Research; Risk Assessment; Role; Signal Transduction; Signaling Protein; Source; Spin Trapping; Structure of parenchyma of lung; Superoxides; Techniques; Testing; Transfection; Transgenic Mice; Tumor Promotion; activating transcription factor; cancer risk; carcinogenesis; catalase; cell transformation; in vivo; nuclear factors of activated T-cells; overexpression; response; transcription factor; tumorigenesis

DESCRIPTION (provided by applicant): Nickel-containing compounds are human carcinogens. The mechanisms of their carcinogenic actions remain to be investigated. Recent studies have indicated that reactive oxygen species (ROS) may play an important role. We hypothesize that nickel induces generation of ROS, which activate nuclear transcription factors, leading to cell transformation and tumorigenesis. Specific Aim 1 will detect and identify ROS generated in human bronchial epithelial cells (BEAS-2B) and mouse embryo fibroblast cells exposed to nickel compounds and investigate the mechanism involved. We hypothesize that nickel (Ni3S2 and NiCl2) can stimulate the cells to activate NADPH oxidase via cdc42 and p47phox to produce superoxide radical, which is then converted to hydrogen peroxide and hydroxyl radical. Specific Aim 2 will test the hypothesis that ROS are required for activation of NFAT and NFkappaB in cells and in vivo in response to nickel compounds. The role of ROS in nickel-induced activation of NFAT and NFkappaB in cells will be evaluated by co-transfection of NFkappaB-luciferase or NFAT-luciferase reporter plasmids and specific ROS scavenger enzymes. For in vivo study, BALB/c transgenic mice with alternation of antioxidant enzymes or NADPH oxidase (ROS generating enzyme) will be employed. Specific Aim 3 will investigate the role of ROS, NFAT and NFkappaB in nickel-induced cell transformation and tumorigenesis. We hypothesize that ROS activate transcription factors and cause cell transformation and tumorigenesis. We will use overexpression of DN-NFAT, DN-IkappaBalpha and DN- IKKbeta to investigate the involvement of NFAT and NFkappaB in nickel-cell transformation and induced tumorigenesis. The role of ROS will be investigated using specific antioxidant enzyme expressions and NADPH oxidase alternation. We anticipate that nickel causes activation of NFAT and NFkappaB through ROS reactions, leading to cell transformation and tumorigenesis. We attempt to link the cell transformation and tumorigenesis with specific transcription factors and specific reactive oxygen species. The results obtained from this proposal will elucidate the role of ROS and NFAT/NFkappaB signaling in Ni compounds-induced carcinogenesis. The long term goals are to provide a fundamental understanding concerning the mechanism of carcinogenic actions of Ni; to fill a need for the mechanistic information of cancer risk assessment for exposure; to propose methods for early detection; and to develop intervention and prevention strategies. Nickel-containing compounds are human carcinogens. This project will investigate the mechanism of Ni-induced carcinogenesis by testing the hypothesis that nickel induces generation of ROS, which activate nuclear transcription factors, leading to cell transformation and tumorigenesis. The long term goals are to understand the mechanism of Ni-carcinogenesis; to propose methods for early detection; and to develop intervention and prevention strategies.

描述（由申请方提供）：含镍化合物是人类致癌物。其致癌作用的机制仍有待研究。最近的研究表明，活性氧（ROS）可能起着重要的作用。我们假设镍诱导产生活性氧，激活核转录因子，导致细胞转化和肿瘤发生。Specific Aim 1将检测和鉴定暴露于镍化合物的人支气管上皮细胞（BEAS-2B）和小鼠胚胎成纤维细胞中产生的ROS，并研究相关机制。我们推测镍（Ni 3S 2和NiCl 2）可以通过cdc 42和p47 phox激活NADPH氧化酶产生超氧阴离子自由基，然后将其转化为过氧化氢和羟基自由基。具体目标2将检验ROS是响应于镍化合物在细胞中和体内激活NFAT和NF κ B所需的假设。将通过共转染NF κ B-荧光素酶或NFAT-荧光素酶报告质粒和特异性ROS清除酶来评估ROS在细胞中镍诱导的NFAT和NF κ B活化中的作用。对于体内研究，将使用具有抗氧化酶或NADPH氧化酶（ROS产生酶）交替的BALB/c转基因小鼠。具体目标3将研究ROS、NFAT和NF κ B在镍诱导的细胞转化和肿瘤发生中的作用。我们推测活性氧激活转录因子，导致细胞转化和肿瘤发生。我们将使用DN-NFAT、DN-IKB α和DN-IKK β的过表达来研究NFAT和NF κ B在镍细胞转化和诱导的肿瘤发生中的参与。ROS的作用将使用特定的抗氧化酶的表达和NADPH氧化酶的交替进行研究。我们预期镍通过ROS反应引起NFAT和NF κ B的活化，从而导致细胞转化和肿瘤发生。我们试图将细胞转化和肿瘤发生与特异性转录因子和特异性活性氧联系起来。从这个建议获得的结果将阐明活性氧和NFAT/NF κ B信号在镍化合物诱导的致癌作用。长期的目标是提供一个基本的了解机制的致癌作用的镍，以满足需要的机械信息的癌症风险评估暴露，提出方法，早期检测，并制定干预和预防策略。含镍化合物是人类致癌物。本项目将通过验证镍诱导ROS的产生，激活核转录因子，导致细胞转化和肿瘤发生的假设来研究镍诱导的致癌机制。长期目标是了解镍致癌的机制;提出早期检测的方法;并制定干预和预防策略。