Cigarette smoke extract reduces FOXO3a promoting tumor progression and cell migration in lung cancer
Cigarette smoke extract reduces FOXO3a promoting tumor progression and cell migration in lung cancer
复制标题
香烟烟雾提取物降低FOXO3a促进肺癌的肿瘤进展和细胞迁移
DOI：
10.1016/j.tox.2021.152751
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发表时间：
2021-03-20
期刊：
影响因子：
4.5
通讯作者：
Pace, Elisabetta
中科院分区：
文献类型：
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作者：
Di Vincenzo, Serena;Sangiorgi, Claudia;Pace, Elisabetta
肺癌是世界范围内癌症死亡的主要原因，烟草烟雾中的致癌物质在其进展和转移中起着重要作用。相关的分子事件在很大程度上是未知的。FOXO3a是一种被认为是肿瘤抑制因子的转录因子。本研究旨在研究香烟烟雾对不同类型肺癌细胞(A549、Colo 699 N、SK-MES-1)线粒体状态和细胞代谢的影响，以及对参与肿瘤进展和细胞迁移的关键途径的影响，探讨FOXO3a在这些机制中的作用。采用流式细胞术、荧光分光光度法、免疫印迹分析、实时定量聚合酶链式反应和划痕试验等方法检测细胞上皮向间充质转化(EMT)标志物(E-cadherin、SNAIL-1)、基质金属蛋白酶-9(MMP9)的表达及细胞迁移情况。结果表明：(1)CSE可增加细胞内ROS、线粒体超氧化物歧化和乳酸释放，降低细胞内ATP含量；(2)减少FOXO3a，增加胞浆中Survivin和p21的表达；(3)减少E-钙粘素，增加SNAIL1和MMP9，促进细胞迁移，与转化生长因子-β1一样。这些作用可以通过下调FOXO3a的表达来部分解释，沉默实验证明了这一点。这些数据表明，香烟烟雾诱导氧化应激和线粒体损伤，导致代谢重新编程与糖酵解通量增加相关。这伴随着FOXO3a的下调，有助于EMT过程和细胞迁移，从而促进肿瘤进展。
Lung cancer is the leading cause of cancer death worldwide, and the carcinogens in tobacco smoke play a role in its progression and metastasis. The related molecular events are largely unknown. FOXO3a is a transcription factor considered a tumor suppressor. Its inhibition leads to cell transformation, tumor progression and metastasis.The aim of this study was to investigate, in different types of lung cancer cell lines (A549, COLO 699 N, SK-MES-1), the effects of cigarette smoke on mitochondrial status and cell metabolism and on key pathways involved in tumor progression and cell migration, looking at the role of FOXO3a in these mechanisms.The different lung cancer cells were exposed to cigarette smoke extract (CSE) and TGF-beta 1. Reactive oxygen species (ROS), mitochondrial superoxide, intracellular ATP, extracellular lactate, FOXO3a, p21, survivin, epithelial-to-mesenchymal transition (EMT) markers (E-cadherin, SNAIL1), MMP-9 and cellular migration were assessed by flow-cytometry, fluorimetry, western blot analysis, Real-Time PCR and scratch test.Our results showed that exposure to CSE: (i) increased ROS, mitochondrial superoxide, lactate release while reducing intracellular ATP; (ii) decreased FOXO3a and increased survivin and p21 in the cytoplasm; (iii) decreased E-cadherin, increased SNAIL1 and MMP-9 and promoted cell migration like TGF-beta 1 did. These effects could be partly explained by downregulation of FOXO3a, as demonstrated by silencing experiments.These data suggest that cigarette smoke induces oxidative stress and mitochondrial damage leading to metabolic reprogramming associated with increased glycolytic flux. This is accompanied with a downregulation of FOXO3a contributing to EMT processes and cell migration therefore promoting tumor progression.