Cytoplasmic Damage and Genotoxicity

细胞质损伤和遗传毒性

• 批准号: 7218072
• 负责人: Tom K. Hei
• 金额: $ 32.91万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-03 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7218072
• 关键词: 4 hydroxynonenal; Acridines; Address; Allopurinol; Alpha Particles; Antibodies; Aromatic Polycyclic Hydrocarbons; Biological; Biological Assay; Cell Communication; Cell Nucleus; Cell Respiration; Cell hybridization; Cells; Charge; Chemicals; Chinese Hamster Ovary Cell; Chromatids; Chromosomes; Chromosomes, Human, Pair 11; Commercial Sources; Communication; Complement; Connexin 43; Cytoplasm; DNA; Data; Deoxyguanosine; Dimethyl Sulfoxide; Dominant-Negative Mutation; Dose; Dyes; Electron Spin Resonance Spectroscopy; End Point; Environmental Carcinogens; Epithelial; Epithelial Cells; Event; Fluorochrome; Frequencies; Gap Junctions; Generations; Genetic; Glutathione; Goals; HPRT1 gene; Hamsters; Human; Hybrid Cells; Hybrids; Hydrogen Peroxide; Hydroxyl Radical; Hypoxanthine Phosphoribosyltransferase; Incidence; Induced Mutation; Lindane; Lipid Peroxidation; Localized; Mammalian Cell; Mediating; Mitochondria; Molecular; Mutagenesis; Mutation; Nature; Nuclear; Numbers; Pathologic Mutagenesis; Play; Population; Principal Investigator; Process; Radial; Radon; Reactive Nitrogen Species; Reactive Oxygen Species; Reporting; Research; Research Personnel; Respiratory Chain; Role; Score; Series; Sister Chromatid Exchange; Source; Spin Trapping; Staining method; Stains; Superoxides; System; Techniques; Tissues; Tocopherols; Toluene; Uridine; Vitamin E; base; cytotoxicity; ditercalinium; genotoxicity; inhibitor/antagonist; irradiation; monolayer; mutant; oxidative DNA damage; particle; programs; research study; response; vector control

DESCRIPTION (provided by applicant): The biological consequences of cytoplasmic damage are largely unknown. The prevailing dogma considered the genotoxic effects of environmental carcinogens such as polycyclic aromatic hydrocarbons and radon alpha particles as being due mostly to direct damage to the nucleus. Using a precision charged particle microbeam and dual fluorochrome dyes to locate nucleus and cellular cytoplasm respectively, thereby avoiding inadvertent traversal of nuclei, the applicant has shown previously that cytoplasmic irradiation is, in fact, mutagenic at the CD59 locus of human-hamster hybrid (AL) cells while inflicting minimal cytotoxicity. Furthermore, preliminary evidence suggests that reactive oxygen species mediate this process. This raised the following questions: What types of oxyradicals are involved and what are their origins? Does this radical generating process involve mitochondrial damage? Do the mutations induced by targeted cytoplasmic irradiation occur in human bronchial epithelial cells (target tissues of environmental radon) as well? And finally, can cytoplasmic irradiation induce bystander mutagenic effect in mammalian cells in a manner similar to what the applicant has recently demonstrated with nuclear traversal of the AL cells. To address these issues, a series of 5 specific aims are proposed to address the 4 testable hypotheses. Mutations will be scored at the CD59 locus of the AL cells and at the HPRT locus in primary human bronchial epithelial cells. The proposed studies will help to address the mechanisms of how cytoplasmic irradiation results in a genetic event in the nucleus. Together with the bystander mutagenic effect, the study will address some of the fundamental issues regarding extranuclear target and how cytoplasmic damages are being processed in mammalian cells.

描述（由申请人提供）：细胞质损伤的生物学后果在很大程度上未知。盛行的教条认为环境致癌物（如多环芳烃和raαα颗粒）的遗传毒性作用主要是由于对核的直接损害。使用精确的电荷颗粒微膜和双荧光组染料分别定位核和细胞胞质，从而避免了核的无意间遍历，先前申请人表明，在人类汉斯特杂交（Al）Cytit（Al）cytit的CD59基因座上，实际上是细胞辐射，实际上是诱变的。此外，初步证据表明，活性氧介导了这一过程。这提出了以下问题：涉及哪些类型的草种，它们的起源是什么？这种根治性生成过程是否涉及线粒体损伤？人支气管上皮细胞（环境ra的靶组织）是否也会发生靶向细胞质辐照引起的突变？最后，细胞质照射可以以类似于申请人最近与Al细胞的核遍历相似的方式诱导哺乳动物细胞中的旁观者诱变作用。为了解决这些问题，提出了一系列5个具体目标来解决这4个可检验的假设。突变将在AL细胞的CD59基因座和原代人支气管上皮细胞中的HPRT基因座进行评分。拟议的研究将有助于解决细胞质照射如何导致细胞核中的遗传事件的机制。该研究将与旁观者的诱变作用一起解决有关核外靶标以及哺乳动物细胞中如何处理细胞质损害的一些基本问题。