Cigarette smoke and asbestos activate poly-ADP-ribose polymerase in alveolar epithelial cells
Cigarette smoke and asbestos activate poly-ADP-ribose polymerase in alveolar epithelial cells
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DOI：
10.2310/6650.2001.34092
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发表时间：
2001-01-01
影响因子：
2.6
通讯作者：
Weitzman, SA
中科院分区：
文献类型：
--
作者：
Kamp, DW;Srinivasan, M;Weitzman, SA
背景：香烟烟雾以协同方式增强石棉诱发的支气管肺癌，其机制尚未确定。一个重要的机制可能涉及氧化剂诱导的DNA损伤导致的肺泡上皮细胞（AEC）损伤，随后激活聚（ADP-核糖）聚合酶（PARP），一种参与DNA修复的酶，可以耗尽细胞能量储存。我们以前表明，整个水香烟烟雾提取物（CSE）增加铁石棉诱导的DNA损伤和细胞毒性培养AEC的一部分，通过产生铁诱导的自由基。我们假设，CSE增加石棉诱导的AEC损伤触发PARP激活导致的DNA损伤引起的铁诱导的freedia.Methods：水CSE新鲜制备的每一个实验的当天。通过在4小时内摄取标记的NAD来评估WI-26（I型样细胞系）和A549（II型样细胞系）细胞中的PARP活性，并基于PARP抑制剂3-氨基苯甲酰胺（3-阿坝）存在下PARP水平的降低来确认。结果：过氧化氢（H2 O2; 1-250 μ M）、CSE（0.4-10 vol%）和铁石棉（5-250 mug/cm（2））均能激活WI-26和A549细胞中的PARP。石棉（5 μ g/cm（2））和CSE（0.04- 10%）的组合诱导WI-26和A549细胞PARP活化，没有协同作用的证据。3-阿坝显著减弱了H2 O2、CSE和石棉对WI-26和A549细胞PARP活性的影响，植酸、铁螯合剂、过氧化氢酶和超氧化物歧化酶均能降低石棉和CSE对WI-26细胞PARP的激活作用。CSE和石棉以非协同方式诱导培养AEC中的PARP活化，这些数据进一步支持石棉和香烟烟雾对相关肺靶细胞具有遗传毒性，铁诱导的自由基部分导致这些效应。
Background: Cigarette smoke augments asbestos-induced bronchogenic carcinoma in a synergistic manner by mechanisms that are not established. One important mechanism may involve alveolar epithelial cell (AEC) injury resulting from oxidant-induced DNA damage that subsequently activates poly (ADP-ribose) polymerase (PARP), an enzyme involved in DNA repair that can deplete cellular energy stores. We previously showed that whole aqueous cigarette smoke extracts (CSE) augment amosite asbestos-induced DNA damage and cytotoxicity to cultured AEC in part by generating iron-induced free radicals. We hypothesized that CSE increase asbestos-induced AEC injury by triggering PARP activation resulting from DNA damage caused by iron-induced free radicals.Methods: Aqueous CSE were prepared fresh on the day of each experiment. PARP activity in WI-26 (a type I-like cell line) and A549 (a type II-like cell line) cells was assessed by the uptake of labeled NAD over 4 hours and confirmed on the basis of the reduction of PARP levels in the presence of a PARP inhibitor, 3-aminobenzamide (3-ABA). Cell survival was assessed by trypan blue dye exclusion.Results: Hydrogen peroxide (H2O2; 1-250 muM), CSE (0.4-10 vol%), and amosite asbestos (5-250 mug/cm(2)) each caused PARP activation in WI-26 and A549 cells. The combination of asbestos (5 mug/cm(2)) and CSE (0.04-10 %) induced WI-26 and A549 cell PARP activation without evidence of synergism. 3-ABA significantlyattenuatedWI-26 and A549 cell PARP activity and cell death after exposure to H2O2, CSE, and asbestos, Phytic acid, an iron chelator, catalase, and superoxide dismutase each decreased WI-26 cell PARP activation caused by asbestos and CSE.Conclusions: CSE and asbestos induce PARP activation in cultured AEC in a nonsynergistic manner, These data provide further support that asbestos and cigarette smoke are genotoxic to relevant lung target cells and that iron-induced free radicals in part cause these effects.