Cardiac glycoside sensitized hepatocellular carcinoma cells to TRAIL via ROS generation, p38MAPK, mitochondrial transition, and autophagy mediation
Cardiac glycoside sensitized hepatocellular carcinoma cells to TRAIL via ROS generation, p38MAPK, mitochondrial transition, and autophagy mediation
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DOI：
10.1002/mc.23096
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发表时间：
2019-08-08
影响因子：
4.6
通讯作者：
Park, Sang-Youel
中科院分区：
文献类型：
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作者：
Rasheduzzaman, Mohammad;Yin, Honghua;Park, Sang-Youel
肿瘤坏死因子相关凋亡诱导配体（TRAIL）在肿瘤中的临床应用的主要问题是耐药性的发展。因此，可以潜在地恢复TRAIL敏感性的药剂是癌症治疗的重要治疗靶标。在此，我们评估了毛花苷c和地高辛，这两者都是广泛使用的强心苷（CG），其通过TRAIL诱导的凋亡对人肝细胞癌细胞（Huh-7和HepG 2）的敏化能力。CG使TRAIL功能化，如其对细胞内活性氧（ROS）产生的影响所示，这损害线粒体完整性，从而在组合治疗期间赋予内在的凋亡半胱天冬酶级联。半胱天冬酶激活依赖于ROS，如CG产生ROS的能力和ROS-N-乙酰半胱氨酸（NAC）关系所示，其通过阻止半胱天冬酶-8和-3的形成来抑制共处理期间的细胞凋亡。此外，CG触发p38 MAPK磷酸化，NAC预暴露阻断p38 MAPK磷酸化，这表明p38 MAPK依赖于ROS的产生。此外，发现CG是AMPK介导的保护性自噬的有效诱导剂，因为药理学和遗传性自噬抑制达到TRAIL介导的细胞凋亡的较高阈值。最后，CG下调抗凋亡蛋白Bcl-2的表达，并增加促凋亡蛋白细胞色素c的易位，从而诱导细胞凋亡。总的来说，这些结果表明，CG通过ROS生成、p38 MAPK磷酸化、细胞存活蛋白下调和保护性自噬抑制增强对TRAIL的细胞毒性能力。
A major concern in the clinical application of tumor necrosis factor related apoptosis-inducing ligand (TRAIL) in tumors is the development of resistance. Therefore, agents that can potentially restore TRAIL sensitivity are important therapeutic targets for cancer treatment. Herein, we evaluated lanatoside c and digoxin, both of which are widely used cardiac glycosides (CGs), for their ability to sensitize human hepatocellular carcinoma cells (Huh-7 and HepG2) through TRAIL-induced apoptosis. CGs functionalize TRAIL as shown by its effect on intracellular reactive oxygen species (ROS) generation, which damages mitochondrial integrity and thereby confers intrinsic apoptotic caspase cascade during combined treatment. Caspase activation is dependent on ROS as shown by the ability of CGs to generate ROS and the ROS-N-acetylcysteine (NAC) relationship, which inhibits apoptosis during cotreatment by preventing the formation of caspase-8 and -3. Furthermore, CGs triggered p38MAPK phosphorylation and NAC pre-exposure blocked p38MAPK phosphorylation, which demonstrated that p38MAPK was dependent upon ROS generation. Additionally, CGs were found to be potent inducers of AMPK-mediated protective autophagy as pharmacological and genetic autophagy inhibition reached the higher threshold of TRAIL-mediated apoptosis. Finally, CGs downregulated the expression of the antiapoptotic protein Bcl-2 and increased the translocation of proapoptotic protein cytochrome c, thereby inducing apoptosis. Collectively, these results indicate that CGs potentiate the enhanced cytotoxic capacity to TRAIL through ROS generation, p38MAPK phosphorylation, cell survival protein downregulation, and protective autophagy inhibition.