Activation of JNK and IRE1 is critically involved in tanshinone I-induced p62 dependent autophagy in malignant pleural mesothelioma cells: implication of p62 UBA domain.
Activation of JNK and IRE1 is critically involved in tanshinone I-induced p62 dependent autophagy in malignant pleural mesothelioma cells: implication of p62 UBA domain.
复制标题
DOI：
10.18632/oncotarget.15336
复制
发表时间：
2017-04-11
期刊：
影响因子：
--
通讯作者：
Kim SH
中科院分区：
文献类型：
--
作者：
Lee J;Sohn EJ;Yoon S;Won G;Kim CG;Jung JH;Kim SH
本研究的目的是阐明丹参酮 I (Tan I) 在 H28 和 H2452 间皮瘤细胞中的自噬机制。在此，Tan I 通过自噬蛋白 5 (ATG5)/微管相关蛋白 1A/1B-轻链 3II (LC3 II) 激活、p62/sequestosome 1 (SQSTM1) 积累以及 LC3II 点、吖啶橙染色细胞和自噬空泡数量增加的自噬特征发挥细胞毒性。然而，3-甲基腺嘌呤 (3MA) 和 NH4Cl 增加了 Tan I 处理的 H28 细胞的细胞毒性。此外，在用 RFP-GFP-LC3 构建体转染的 Tan I 处理的 H28 细胞中，自噬流增强，并且 GFP-LC3 点与 LAMP1 或 Lysotracker 共定位。有趣的是，C 端 UBA 结构域是 Tan 1 诱导 H28 细胞中 p62 聚集所必需的。值得注意的是，Tan I 上调了 CCAAT 增强子结合蛋白同源蛋白 (CHOP)、肌醇需要蛋白 1 (IRE1) 和 p-c-Jun N 末端激酶 (p-JNK)，但 IRE1 或 p62 和 JNK 抑制剂 SP600125 的沉默阻止了 Tan I 处理的 H28 细胞中 LC3II 的积累。总的来说，这些发现表明 Tan I 通过在恶性间皮瘤细胞中激活 JNK 和 IRE 1，在细胞凋亡和 p62/SQSTM1 依赖性自噬之间进行折衷，从而发挥抗肿瘤活性。
The aim of present study is to elucidate autophagic mechanism of tanshinone I (Tan I) in H28 and H2452 mesothelioma cells. Herein, Tan I exerted cytotoxicity with autophagic features of autophagy protein 5 (ATG5)/ microtubule-associated protein 1A/1B-light chain 3II (LC3 II) activation, p62/sequestosome 1 (SQSTM1) accumulation and increased number of LC3II punctae, acridine orange-stained cells and autophagic vacuoles. However, 3-methyladenine (3MA) and NH4Cl increased cytotoxicity in Tan I treated H28 cells. Furthermore, autophagy flux was enhanced in Tan I-treated H28 cells transfected by RFP-GFP-LC3 constructs, with colocalization of GFP-LC3 punctae with LAMP1 or Lysotracker. Interestingly, C-terminal UBA domain is required for Tan 1 induced aggregation of p62 in H28 cells. Notably, Tan I upregulated CCAAT-enhancer-binding protein homologous protein (CHOP), inositol-requiring protein-1 (IRE1) and p-c-Jun N-terminal kinase (p-JNK), but silencing of IRE1 or p62 and JNK inhibitor SP600125 blocked the LC3II accumulation in Tan I-treated H28 cells. Overall, these findings demonstrate that Tan I exerts antitumor activity through a compromise between apoptosis and p62/SQSTM1-dependent autophagy via activation of JNK and IRE 1 in malignant mesothelioma cells.