Compound C sensitizes Caki renal cancer cells to TRAIL-induced apoptosis through reactive oxygen species-mediated down-regulation of c-FLIPL and Mcl-1
Compound C sensitizes Caki renal cancer cells to TRAIL-induced apoptosis through reactive oxygen species-mediated down-regulation of c-FLIPL and Mcl-1
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DOI：
10.1016/j.yexcr.2010.04.028
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发表时间：
2010-08-01
影响因子：
3.7
通讯作者：
Kwon, Taeg Kyu
中科院分区：
文献类型：
--
作者：
Jang, Ji Hoon;Lee, Tae Jin;Kwon, Taeg Kyu
肿瘤坏死因子相关凋亡诱导配体（TRAIL）无论是单独使用还是与其他抗癌药物联合使用，都被认为是抗癌治疗的新策略。化合物 C 是一种细胞渗透性吡唑并嘧啶衍生物，可作为 AMP 激活蛋白激酶 (AMPK) 的有效、选择性、可逆 ATP 竞争性抑制剂。在这项研究中，我们发现化合物 C 使 Caki 人肾癌细胞对 TRAIL 介导的细胞凋亡敏感，但对正常人皮肤成纤维细胞 (HSF) 和人系膜细胞不敏感。然而，AMPK siRNA 未能影响 Caki 细胞中 TRAIL 介导的细胞凋亡，显性失活 AMPK 的转导反而减弱了 TRAIL 诱导的细胞凋亡，表明化合物 C 对 TRAIL 诱导的细胞凋亡的敏化作用与 AMPK 活性无关。有趣的是，我们发现 c-FLIPL 和 Mcl-1 的下调有助于化合物 C 增强 TRAIL 诱导的细胞凋亡。在化合物 C 处理的细胞中，c-FLIPL 和 Mcl-1 表达的降低是由 c-FLIPL 和 Mcl-1 的蛋白质稳定性降低引起的，而不是由它们的转录控制引起的。 N-乙酰-L-半胱氨酸（NAC）预处理显着抑制了化合物C和TRAIL联合治疗诱导的细胞死亡，并恢复了化合物C加TRAIL联合治疗下调的c-FLIPL和Mcl-1的表达水平，表明化合物C刺激的TRAIL诱导的细胞凋亡似乎依赖于活性氧的产生来下调c-FLIPL和 Mcl-1。综上所述，本研究表明，化合物 C 通过 ROS 介导的 c-FLIPL 和 Mcl-1 下调，增强 TRAIL 诱导的人肾癌细胞凋亡。 (C) 2010 Elsevier Inc. 保留所有权利。
The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), either alone or in combination with other anticancer drugs, is considered as a new strategy for anticancer therapy. Compound C, a cell-permeable pyrrazolopyrimidine derivative, acts as a potent, selective, reversible ATP-competitive inhibitor of AMP-activated protein kinase (AMPK). In this study, we show that compound C sensitizes Caki human renal cancer cells, but not normal human skin fibroblast cells (HSF) and human mesangial cells, to TRAIL-mediated apoptosis. However, AMPK siRNA failed to affect TRAIL-mediated apoptosis in Caki cells and transduction of dominant negative AMPK rather attenuated TRAIL-induced apoptosis, indicating that the effect of compound C on sensitization of TRAIL-induced apoptosis is independent of AMPK activity. Interestingly, we found that down-regulation of c-FLIPL and Mcl-1 contributes to compound C-enhanced TRAIL-induced apoptosis. Reduced expression of c-FLIPL and Mcl-1 were caused by the decreased protein stability of c-FLIPL and Mcl-1, but not by their transcriptional control, in compound C-treated cells. Pretreatment with N-acetyl-L-cysteine (NAC) significantly inhibited the cell death induced by the combined treatment with compound C and TRAIL as well as recovered the expression levels of c-FLIPL and Mcl-1 down-regulated by the combinatory treatment with compound C plus TRAIL, suggesting that compound C-stimulated TRAIL-induced apoptosis appears to be dependent on the generation of reactive oxygen species for down-regulation of c-FLIPL and Mcl-1. Taken together, the present study demonstrates that compound C enhances TRAIL-induced apoptosis in human renal cancer cells by ROS-mediated c-FLIPL and Mcl-1 down-regulation. (C) 2010 Elsevier Inc. All rights reserved.