Dichloroacetate induces protective autophagy in LoVo cells: involvement of cathepsin D/thioredoxin-like protein 1 and Akt-mTOR-mediated signaling.
Dichloroacetate induces protective autophagy in LoVo cells: involvement of cathepsin D/thioredoxin-like protein 1 and Akt-mTOR-mediated signaling.
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二氯乙酸诱导 LoVo 细胞中的保护性自噬：组织蛋白酶 D/硫氧还蛋白样蛋白 1 和 Akt-mTOR 介导的信号传导的参与
DOI：
10.1038/cddis.2013.438
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发表时间：
2013-11-07
影响因子：
9
通讯作者：
中科院分区：
文献类型：
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作者：
二氯乙酸（DCA）是丙酮酸脱氢酶激酶（PDK）的抑制剂，近年来已被证明是一种很有前途的无毒抗肿瘤药物。在这项研究中，我们证明了DCA可以诱导LoVo细胞的自噬，这是通过自噬体的形成，LC 3免疫反应性的点状图案的出现和自噬相关蛋白的激活来证实的。此外，通过3-甲基腺嘌呤（3-MA）或Atg 7 siRNA处理的自噬抑制可以显著增强DCA诱导的细胞凋亡。为了确定DCA诱导的自噬的潜在机制，使用药物亲和力响应性靶稳定性（DARTS）结合ESI-Q-TOF MS/MS分析的靶鉴定用于描绘对照和DCA处理的LoVo细胞之间差异表达的蛋白质。结果，与对照组相比，组织蛋白酶D（CTSD）和硫氧还蛋白样蛋白1（TXNL 1）发生了显着变化。进一步的研究表明，DCA处理显着促进异常活性氧（ROS）的产生。另一方面，活性氧抑制剂N-乙酰半胱氨酸（NAC）可以减弱DCA引发的自噬。最后，我们证明了Akt-mTOR信号通路，自噬的主要负调节因子，被DCA处理抑制。据我们所知，这是第一个研究表明DCA诱导LoVo细胞保护性自噬，其潜在机制涉及ROS失衡和Akt-mTOR信号通路抑制。
Dichloroacetate (DCA) is an inhibitor of pyruvate dehydrogenase kinase (PDK), and recently it has been shown as a promising nontoxic antineoplastic agent. In this study, we demonstrated that DCA could induce autophagy in LoVo cells, which were confirmed by the formation of autophagosomes, appearance of punctate patterns of LC3 immunoreactivity and activation of autophagy associated proteins. Moreover, autophagy inhibition by 3-methyladenine (3-MA) or Atg7 siRNA treatment can significantly enhance DCA-induced apoptosis. To determine the underlying mechanism of DCA-induced autophagy, target identification using drug affinity responsive target stability (DARTS) coupled with ESI-Q-TOF MS/MS analysis were utilized to profile differentially expressed proteins between control and DCA-treated LoVo cells. As a result, Cathepsin D (CTSD) and thioredoxin-like protein 1 (TXNL1) were identified with significant alterations compared with control. Further study indicated that DCA treatment significantly promoted abnormal reactive oxygen species (ROS) production. On the other hand, DCA-triggered autophagy could be attenuated by N-acetyl cysteine (NAC), a ROS inhibitor. Finally, we demonstrated that the Akt-mTOR signaling pathway, a major negative regulator of autophagy, was suppressed by DCA treatment. To our knowledge, it was the first study to show that DCA induced protective autophagy in LoVo cells, and the potential mechanisms were involved in ROS imbalance and Akt-mTOR signaling pathway suppression.
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影响因子：
8.8
作者：
Michelakis, E. D.;Webster, L.;Mackey, J. R.
通讯作者：
Mackey, J. R.
影响因子：
4.8
作者：
Haendeler, J;Popp, R;Dimmeler, S
通讯作者：
Dimmeler, S
影响因子：
4.7
作者：
Lin, Ji-Fan;Tsai, Te-Fu;Hwang, Thomas I-Sheng
通讯作者：
Hwang, Thomas I-Sheng
DOI：
10.1083/jcb.200412022
发表时间：
2005-05-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Komatsu M;Waguri S;Ueno T;Iwata J;Murata S;Tanida I;Ezaki J;Mizushima N;Ohsumi Y;Uchiyama Y;Kominami E;Tanaka K;Chiba T
通讯作者：
Chiba T
影响因子：
15.9
作者：
McMurtry, MS;Archer, SL;Michelakis, ED
通讯作者：
Michelakis, ED