Pathways of PAH activation in human lung cells

人肺细胞中 PAH 激活途径

基本信息

批准号：
7478339
负责人：
Trevor M Penning
金额：
$ 47.65万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-03 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7478339
关键词：
8-Oxo-2&apos -Deoxyguanosine A549 Adenocarcinoma Cell Alleles Antibodies Aromatic Hydrocarbons Aromatic Polycyclic Hydrocarbons Aryl Hydrocarbon Receptor Attenuated Benzo(a)pyrene Binding Biological Assay Biological Monitoring Breathing Cancer Patient Candidate Disease Gene Carcinogens Cations Cell Line Cell model Cells Collaborations Coupled Cytochrome P450 DNA DNA Adducts DNA Damage Data Deoxyguanosine Environmental Pollutants Environmental Tobacco Smoke Enzyme Induction Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Epithelial Epithelial Cells Epoxy Compounds Excision Fossil Fuels Genes Genetic Predisposition to Disease Genetic Transcription Glycol Hand Human In Vitro Institutes International Agency for Research on Cancer Label Laboratories Link Liquid Chromatography Lung Malignant Epithelial Cell Malignant neoplasm of lung Measurement Measures Mediating Metabolic Metabolic Activation Metabolism Methodology Methods Modeling Monitor Normal Cell Oxidation-Reduction Particulate Matter Pathway interactions Patients Pennsylvania Peroxidase Peroxidases Phenotype Polycyclic Hydrocarbons Predisposition Protein Isoforms Protocols documentation Quinones Radioisotope Dilution Technique Reaction Receptor Activation Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Route Small Interfering RNA Smoke Standards of Weights and Measures Sulforaphane Telomerase Testing Tetrachlorodibenzodioxin Time Tobacco smoke Toxic effect Universities World Health Organization adduct anticancer research base benzo(a)pyrene tetrol benzoquinone carbonyl reductase (NADPH)cell transformation exhaust immortalized cell in vivo inhibitor/antagonist knock-down oxidative DNA damage pollutant programs repair enzyme stable isotope tandem mass spectrometry

项目摘要

DESCRIPTION (provided by applicant): Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants that have been upgraded to human carcinogens by the WHO, International Agency for Research on Cancer (IARC). PAH are airborne pollutants found in smoke from fossil-fuel combustion, car-exhaust, fine-particulate matter (PM2.5), and first and second hand tobacco smoke. Inhalation toxicity likely results in cancer of the lung and airway. PAH require metabolic activation to exert their deleterious effects, however, the major pathway of PAH activation in the carcinogen target cell, normal human bronchial epithelial cells (NHBE) is uncertain. Three pathways have been proposed: the radical cation pathway (P450-peroxidase dependent) which produces depurinating-DNA adducts; the diol-epoxide pathway (P450-dependent) which produces stable-DMA adducts; and the formation of reactive and redox-active o-quinones (aldo-keto reductase (AKR) dependent), which can give rise to covalent DMA adducts and oxidative damage of DMA. Based on the consistent over-expression of AKR isoforms and the loss of one allele of hOGG1 (human oxoguanine glycosylase the base excision repair enzyme specific for removal 8-oxo-dGuo from DMA) in patients with lung cancer, we hypothesize that AKRs activate PAHs in human bronchial epithelial (HBE) cells and this results in increased oxidative DNA damage. Four aims are proposed to test this hypothesis. In Aim #1, the metabolism of benzo[a]pyrene (BP) will be measured in parental H358 cells (transformed HBE cells) in the presence and absence of (P450 and AKR) induction and the levels of radical cation metabolites (BP-1,6-, 3,6- and 6,12-diones), diol-epoxide metabolites (BP-tetrols) and o-quinone metabolites (BP-7,8-dione) will be quantified and identified by LC- MS. In Aim#2, a stable isotope dilution method using [13C]-BP-metabolites as internal standards to monitor the BP-metabolome will be established. In Aim#3, NHBE cells immortalized with cdk4 and human telomerase will be used to monitor the BP-metabolome by LC-MS stable isotope dilution methodology in the presence and absence of P450 and AKR inducers. In Aim#4, the ability of BP, BP-7,8-dihydrodiol (AKR substrate) and BP-7,8-dione (AKR product) to cause AKR-dependent 8-oxo-dGuo formation will be examined in NHBE cells. These studies will identify major BP-metabolites in NHBE cells which could be used to biomonitor human PAH exposure, and validate candidate genes for genetic predisposition studies.

描述（由申请人提供）：多环芳烃（PAHS）是环境污染物，已由国际癌症研究机构（IARC）升级为人类致癌物。 PAH是在化石燃料燃烧，避免，精细的物质（PM2.5）以及第一手和二手烟雾中发现的空中污染物。吸入毒性可能导致肺癌和气道癌。 PAH需要代谢激活才能发挥其有害作用，但是，正常的人支气管上皮细胞（NHBE）是致癌靶细胞中PAH激活的主要途径。已经提出了三种途径：产生偏移DNA加合物的自由基阳离子途径（P450-过氧酶依赖性）；产生稳定的DMA加合物的二醇环氧途径（依赖P450）；以及反应性和氧化还原活性的O- Quinones（Aldo-Keto还原酶（AKR）依赖性）的形成，这可以引起共价DMA加合物和DMA的氧化损伤。基于AKR同工型的一致过表达和HOGG1等位基因的丧失（人氧谁糖糖基化酶在肺癌患者中特异性的8-氧DGUO的基本切除修复酶）在肺癌患者中，我们假设AKR在人类bronchial Epithitial（Heble）中均增加了ost of the pahs spain nebne nebna s ne staim spaine nebne nebne nebne ne staignial（hbe）。提出了四个目标来检验这一假设。 In Aim #1, the metabolism of benzo[a]pyrene (BP) will be measured in parental H358 cells (transformed HBE cells) in the presence and absence of (P450 and AKR) induction and the levels of radical cation metabolites (BP-1,6-, 3,6- and 6,12-diones), diol-epoxide metabolites (BP-tetrols) and O-夸酮代谢产物（BP-7,8-二酮）将通过LC-MS量化和鉴定。在AIM＃2中，将建立使用[13C] -bp-企业作为监测BP-METABOLOME的内部标准的稳定同位素稀释方法。在AIM＃3中，在存在和不存在P450和AKR诱导剂的情况下，使用CDK4永生和人端粒酶永生的NHBE细胞将通过LC-MS稳定的同位素稀释方法来监测BP-Metabolome。在AIM＃4中，将在NHBE细胞中检查BP，BP-7,8-二氢二醇（AKR底物）和BP-7,8-二酮（AKR产物）引起AKR依赖性的8-oxo-Dguo形成的能力。这些研究将在NHBE细胞中确定可用于生物监测的人类PAH暴露的主要BP量代谢物，并验证候选基因进行遗传易感研究。