WX20120108, a novel IAP antagonist, induces tumor cell autophagy via activating ROS-FOXO pathway
WX20120108, a novel IAP antagonist, induces tumor cell autophagy via activating ROS-FOXO pathway
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新型IAP拮抗剂WX20120108通过激活ROS-FOXO通路诱导肿瘤细胞自噬
DOI：
10.1038/s41401-019-0253-5
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发表时间：
2019-11-01
影响因子：
8.2
通讯作者：
Wang, Li-li
中科院分区：
文献类型：
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作者：
Ding, Rui;Wang, Xin;Wang, Li-li
最近，发现凋亡蛋白（IAP）和一些IAP拮抗剂的抑制剂调节自噬，但潜在的机制尚不清楚。 WX20120108是GDC-0152（已知IAP拮抗剂）的类似物，在肿瘤细胞中显示出更有效的抗肿瘤和自噬调节活性，我们研究了WX20120108诱导的自噬的调节机制。使用分子对接和荧光极化各向异性（FPA）竞争测定法，我们首先证明，与XIAP-BIR3，XIAP BIR2-BIR3，CIAP1 BIR3和CIAP2 BIR3域的XIAP-BIR3，XIAP BIR2-BIR3，XIAP BIR2-BIR3，CIAP2 BIR3域结合使用的WX20120108。在六个癌细胞系中，WX20120108抑制了细胞的增殖，其效力比GDC-0152高两至十倍。在HELA和MDA-MB-231细胞中，WX20120108诱导caspase依赖性细胞凋亡和激活的TNFA依赖性外部凋亡。另一方面，WX20120108在剂量和时间依赖性的方式中诱导HELA和MDA-MB-231细胞自噬。我们透露，WX20120108在基因修饰细胞系和HELA细胞中的FoxO3核转运以及FOXO3靶向基因的上调表达（BNIP3，PIK3C3，ATG5和ATG4B）的上调表达中，有选择地激活的FOXO3，在自动启动中起着关键作用。当FoxO3基因沉默时，WX201208诱导的自噬会显着抑制。 WX20120108剂量依赖性地增加了HeLa细胞中活性氧（ROS）的产生，而WX20120108诱导的FOXO3激活在存在催化酶（已知的ROS清除剂）的情况下完全阻止。但是，WX201208诱导的ROS产生不受CIAP1/2或XIAP基因沉默的影响。总之，WX201208诱导的自噬依赖于激活与IAP无关的Ros-Foxo3途径。这一发现提供了对IAP拮抗剂介导的自噬调节机制的新见解。
Recently, inhibitor of apoptosis proteins (IAPs) and some IAP antagonists were found to regulate autophagy, but the underlying mechanisms remain unclear. WX20120108 is an analogue of GDC-0152 (a known IAP antagonist) and displays more potent anti-tumor and autophagy-regulating activity in tumor cells, we investigated the regulatory mechanisms underlying WX20120108-induced autophagy. Using molecular docking and fluorescence polarization anisotropy (FPA) competitive assay, we first demonstrated that WX20120108, acting as an IAP antagonist, bound to the XIAP-BIR3, XIAP BIR2-BIR3, cIAP1 BIR3, and cIAP2 BIR3 domains with high affinities. In six cancer cell lines, WX20120108 inhibited the cell proliferation with potencies two to ten-fold higher than that of GDC-0152. In HeLa and MDA-MB-231 cells, WX20120108 induced caspase-dependent apoptosis and activated TNFα-dependent extrinsic apoptosis. On the other hand, WX20120108 induced autophagy in HeLa and MDA-MB-231 cells in dose- and time-dependent manners. We revealed that WX20120108 selectively activated Foxo3, evidenced by Foxo3 nuclear translocation in both gene modified cell line and HeLa cells, as well as the upregulated expression of Foxo3-targeted genes (Bnip3,Pik3c3,Atg5, andAtg4b), which played a key role in autophagy initiation. WX20120108-induced autophagy was significantly suppressed whenFoxo3gene was silenced. WX20120108 dose-dependently increased the generation of reactive oxygen species (ROS) in HeLa cells, and WX20120108-induced Foxo3 activation was completely blocked in the presence of catalase, a known ROS scavenger. However, WX20120108-induced ROS generation was not affected bycIAP1/2orXIAPgene silencing. In conclusion, WX20120108-induced autophagy relies on activating ROS-Foxo3 pathway, which is independent of IAPs. This finding provides a new insight into the mechanism of IAP antagonist-mediated regulation of autophagy.