Autophagic cell death induced by resveratrol depends on the Ca2+/AMPK/mTOR pathway in A549 cells
Autophagic cell death induced by resveratrol depends on the Ca2+/AMPK/mTOR pathway in A549 cells
复制标题
白藜芦醇诱导的自噬细胞死亡依赖于A549细胞中的Ca2/AMPK/mTOR通路
DOI：
10.1016/j.bcp.2013.05.003
复制
发表时间：
2013-07-15
影响因子：
5.8
通讯作者：
Li, Guanwu
中科院分区：
文献类型：
--
作者：
Zhang, Jun;Chiu, Jenfu;Li, Guanwu
白藜芦醇具有抗肿瘤、抗病毒、血管保护等多种生物学作用。最近的研究表明，白藜芦醇通过一种未知的机制诱导自噬发挥其抗肿瘤作用。在这项研究中，我们研究了自噬在白藜芦醇诱导的A549人肺腺癌细胞死亡中的作用及其可能的分子机制。用四甲基偶氮唑盐比色法和集落形成法检测白藜芦醇诱导的细胞生长抑制和细胞死亡。用单烷基硫代巴比妥钠、透射电子显微镜和自噬标记蛋白Lc3的表达来表征自噬的激活。Western印迹分析研究自噬死亡机制中涉及的细胞信号。FURA2-AM染色检测细胞内游离钙离子浓度。我们的结果表明，白藜芦醇诱导的A549细胞死亡是通过自噬介导的。自噬抑制剂3-甲基腺嘌呤抑制白藜芦醇诱导的自噬细胞死亡，并用siRNAs敲除自噬相关基因ATG5和Beclin-1逆转RSV诱导的细胞死亡。加入白藜芦醇后，细胞内游离钙即刻积累，导致磷酸化AMPK和磷酸化Raptor激活，磷酸化p70S6K减少。这些作用可被AMPK抑制剂化合物C和钙离子螯合剂EGTA逆转。综上所述，我们证明白藜芦醇诱导的A549细胞死亡是通过钙/AMPK-mTOR信号通路介导的自噬细胞死亡过程。(C)2013 Elsevier Inc.保留所有权利。
Resveratrol has many biological effects, including anti-tumor, antiviral activities, and vascular protection. Recent studies have suggested that resveratrol exert its antitumor effects through induction of autophagy by an unknown mechanism. In this study, we investigated the involvement of autophagy in resveratrol-induced cell death and its potential molecular mechanisms in A549 human lung adnocarcinoma cells. Resveratrol-induced growth inhibition and cell death was assessed by MTT and clonogenic assays. Activation of autophagy was characterized by monodansylcadaverine, transmission electron microscopy, and expression of autophaky marker protein LC3. Western blot analysis was used to study the cell signals involved in the mechanisms of autophagic death. Intracellular free calcium was detected with Fura2-AM staining. Our results indicated that resveratrol induced A549 cell death was mediated by autophagy. 3-methyladenine, an inhibitor of autophagy, suppressed resveratrol-induced autophagic cell death, and knockdown of autophagy-related genes Atg5 and Beclin-1 with siRNAs reversed RSV-induced cell death. Intracellular free calcium accumulated immediately following resveratrol addition, which led to the activation of phospho-AMPK and phospho-Raptor, and a reduction in the amount of phospho-p70S6K. These effects could be reversed by the AMPK inhibitor compound C, and the calcium ion-chelating agent EGTA. In conclusion, we demonstrate that resveratrol-induced A549 cell death was mediated by the process of autophagic cell death via Ca2+/AMPK-mTOR signaling pathway. (C) 2013 Elsevier Inc. All rights reserved.