MicroRNA-190 and Oxidative Stress in Arsenic carcinogenesis

MicroRNA-190 和砷致癌过程中的氧化应激

基本信息

批准号：
8538383
负责人：
Fei Chen
金额：
$ 33.28万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8538383
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

项目摘要

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.

描述（申请人提供）：我们之前的研究已经清楚地证明三价砷（As3+）可以诱导细胞转化。本申请的目的是确定 miR-190 在 As3+ 诱导的支气管上皮细胞恶性转化和肺部肿瘤发生中的作用。主要重点是 ROS 和 Erk 介导的 miR-190 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+ 以剂量和时间依赖性方式诱导人支气管上皮细胞系 BEAS-2B 和人小气道上皮细胞系 SAEC 中 miR-190 的产生； (b) MiR-190 通过下调 PHLPP（Akt 信号传导的内源性抑制剂和肿瘤抑制因子）的合成来介导 As3+ 诱导的 Akt 激活； (c) miR-190 的过度表达导致细胞转化。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，随后发生细胞转化，从而导致肿瘤发生。为了检验这一假设，提出了三个具体目标：具体目标 1 将研究 As3+ 如何在 BEAS-2B 和 SAEC 细胞系中诱导 miR-190。我们将重点关注 Erk（丝裂原激活蛋白激酶 (MAPK) 家族的关键成员）在 As3+ 处理的细胞中对前体 miR-190 的转录和成熟的调节，具体目标 2 将确定 As3+ 的作用- 诱导活性氧 (ROS) 对 As3+ 激活 Erk 和 miR-190 的产生。我们将鉴定 As3+ 诱导的每种活性氧，并确定 As3+ 诱导的 ROS 的来源，这些 ROS 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，通过过表达 miR-190、抗氧化酶以及 shRNA 介导的 Erk 或 Akt 沉默，研究 miR-190、ROS、Erk 和 Akt 在 As3+ 诱导的癌发生中的作用。人支气管上皮细胞。我们将使用稳定的转染子，并通过软琼脂中不依赖贴壁的生长（集落形成）和接种测定，确定过表达 miR-190、抗氧化酶和 Erk 或 Akt 沉默对基础或 As3+ 诱导的细胞转化和癌变的影响裸鼠肺细胞。该项目的完成将建立As3+诱导miR-190生成的机制及其在As3+致癌作用中的作用。