MicroRNA-190 and Oxidative Stress in Arsenic carcinogenesis

MicroRNA-190 和砷致癌过程中的氧化应激

• 批准号: 8370816
• 负责人: Fei Chen
• 金额: $ 35.04万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8370816
• 关键词: Agar; American; Anchorage-Independent Growth; Animals; Antioxidants; Area; Arsenic; Biogenesis; Biological Assay; Biological Markers; Carcinogenicity Tests; Carcinogens; Case-Control Studies; Cell Line; Cells; Cohort Studies; Country; Data; Dose; Early Diagnosis; Environment; Environmental Exposure; Environmental and Occupational Exposure; Enzymes; Epithelial Cells; Exposure to; Family; General Population; Generations; Genes; Genetic Transcription; Goals; Human; International Agency for Research on Cancer; Intervention; Knowledge; Link; Literature; Lung; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methods; MicroRNAs; Mitochondria; Mitogen-Activated Protein Kinases; Molecular; NADPH Oxidase; Non-Small-Cell Lung Carcinoma; Nude Mice; Occupational Exposure; Oxidative Stress; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Prevention strategy; Production; Protein Biosynthesis; Proteins; Public Health; Reactive Oxygen Species; Regulation; Reporting; Research; Risk; Risk Assessment; Role; Signal Pathway; Signal Transduction; Source; Testing; Time; Transcriptional Regulation; Tumor Suppressor Proteins; United States; base; cancer risk; carcinogenesis; cell growth regulation; cell transformation; human DICER1 protein; inhibitor/antagonist; lung tumorigenesis; member; overexpression; population based; research study; response; small hairpin RNA; tumorigenesis; tumorigenic; Agar; American; Anchorage-Independent Growth; Animals; Antioxidants; Area; Arsenic; Biogenesis; Biological Assay; Biological Markers; Carcinogenicity Tests; Carcinogens; Case-Control Studies; Cell Line; Cells; Cohort Studies; Country; Data; Dose; Early Diagnosis; Environment; Environmental Exposure; Environmental and Occupational Exposure; Enzymes; Epithelial Cells; Exposure to; Family; General Population; Generations; Genes; Genetic Transcription; Goals; Human; International Agency for Research on Cancer; Intervention; Knowledge; Link; Literature; Lung; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methods; MicroRNAs; Mitochondria; Mitogen-Activated Protein Kinases; Molecular; NADPH Oxidase; Non-Small-Cell Lung Carcinoma; Nude Mice; Occupational Exposure; Oxidative Stress; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Prevention strategy; Production; Protein Biosynthesis; Proteins; Public Health; Reactive Oxygen Species; Regulation; Reporting; Research; Risk; Risk Assessment; Role; Signal Pathway; Signal Transduction; Source; Testing; Time; Transcriptional Regulation; Tumor Suppressor Proteins; United States; base; cancer risk; carcinogenesis; cell growth regulation; cell transformation; human DICER1 protein; inhibitor/antagonist; lung tumorigenesis; member; overexpression; population based; research study; response; small hairpin RNA; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): Our previous research has clearly demonstrated that trivalent arsenic (As3+) can induce cell transformation. The goal of this application is to determine the role of miR-190 in As3+-induced malignant transformation of the bronchial epithelial cells and the tumorigenesis of the lung. Major emphasis will be on the ROS- and Erk- mediated expression of miR-190 and the miR-190-dependent Akt activation and tumorigenesis. Our preliminary data have shown that (a) As3+ induces miR-190 generation in the human bronchial epithelial cell line, BEAS-2B, and human small airway epithelial cell line, SAEC, in a manner of both dose- and time- dependent; (b) MiR-190 mediates As3+-induced Akt activation by down-regulating the synthesis of PHLPP, an endogenous inhibitor of Akt signaling and a tumor suppressor; and (c) overexpression of miR-190 causes cell transformation. Based on these preliminary studies, we hypothesize that As3+-induced miR-190 is responsible for the sustained Akt activation, followed by cell transformation and consequently, the tumorigenesis. To test this hypothesis, three specific aims are proposed: Specific Aim 1 will investigate how As3+ induces miR-190 in BEAS-2B and SAEC cell lines. We will focus on the regulation of both transcription and maturation of the precursor miR-190 by Erk, a key member of mitogen-activated protein kinase (MAPK) family, in the cells treated with As3+, Specific Aim 2 will determine the role of As3+-induced reactive oxygen species (ROS) on the activation of Erk and miR-190 production by As3+. We will identify each of the reactive oxygen species induced by As3+, and determine the sources of As3+-induced ROS that activate Erk and induce miR-190; Specific Aim 3 will use overexpression and orthotopic tumorigenesis strategies to study the role of miR-190, ROS, Erk, and Akt in As3+-induced carcinogenesis by overexpressing miR-190, the antioxidant enzymes, and shRNA- mediated silencing of Erk or Akt in the human bronchial epithelial cells. We will use stable transfectants and determine the effects of overexpressing miR-190, antioxidant enzymes, and Erk or Akt silencing on either basal or As3+-induced cell transformation and carcinogenesis by assays of anchorage-independent growth in soft agar (colony formation) and inoculation of the cells in the lung of nude mice. The completion of this project will establish the mechanism of As3+-induced miR-190 generation and its role in As3+ carcinogenesis. PUBLIC HEALTH RELEVANCE: Environmental exposure of arsenic, especially the trivalent form arsenic, has long been a major public health concern. The present study will investigate the mechanism of arsenic-induced carcinogenesis by testing the hypothesis that arsenic-induced microRNA-190 is responsible for the malignant transformation and tumorigenesis of the cells. The long-term goals are to understand molecular mechanism of arsenic-induced carcinogenesis and to identify biomarkers for developing early detection, intervention and prevention strategies.

描述（由申请人提供）：我们以前的研究清楚地表明，三价砷（AS3+）可以诱导细胞转化。该应用的目的是确定miR-190在AS3+诱导的支气管上皮细胞的恶性转化和肺的肿瘤发生中的作用。主要重点是miR-190的ROS和ERK介导的表达以及miR-190依赖性AKT激活和肿瘤发生。我们的初步数据表明，（A）AS3+会以剂量和时间依赖性的方式诱导人支气管上皮细胞系，BEAS-2B和人类小气道上皮细胞系中的miR-190产生。 （b）miR-190通过下调PHLPP的合成（Akt信号传导的内源性抑制剂和肿瘤抑制器），介导AS3+诱导的AKT激活。 （c）miR-190的过表达导致细胞转化。基于这些初步研究，我们假设AS3+诱导的miR-190负责持续的Akt激活，然后是细胞转化，因此是肿瘤发生。为了检验这一假设，提出了三个特定目标：具体目标1将研究AS3+如何在BEAS-2B和SAEC细胞系中诱导miR-190。我们将重点介绍由AS3+处理的细胞中，特定的AIM 2在细胞中，ERK的转录和成熟度（MIRK）的转录和成熟。我们将确定由AS3+诱导的每个活性氧，并确定AS3+诱导的ROS的来源激活ERK并诱导miR-190。具体目标3将使用过表达和原位肿瘤发生策略来研究MiR-190，ROS，ERK和AKT在AS3+诱导的癌发生中的作用，通过过表达MiR-190，抗氧化酶，抗氧化剂酶，ERK或AKT在人体sephithial中的ERK或AKT的shRNA介导的沉积物。我们将使用稳定的转染剂，并通过在软琼脂（菌落形成）中锚固不依赖性的细胞生长的测定法（colony形成）和nude micece nude Miese中的细胞中的细胞（colony形成）中锚固生长的测定，确定过度表达MiR-190，抗氧化酶以及ERK或AKT沉默对基础或AS3+诱导的细胞转化和癌变的影响。该项目的完成将建立AS3+诱导的miR-190世代的机制及其在AS3+癌变中的作用。 公共卫生相关性：砷的环境暴露，尤其是三价形式的砷，长期以来一直是公共卫生的主要问题。本研究将通过检验砷诱导的microRNA-190的假设来研究砷诱导的癌变的机制，负责细胞的恶性转化和肿瘤发生。长期目标是了解砷诱导的致癌作用的分子机制，并确定生物标志物以开发早期检测，干预和预防策略。