BCL2 inhibitor ABT-199 and BCL2L1 inhibitor WEHI-539 coordinately promote NOXA-mediated degradation of MCL1 in human leukemia cells
BCL2 inhibitor ABT-199 and BCL2L1 inhibitor WEHI-539 coordinately promote NOXA-mediated degradation of MCL1 in human leukemia cells
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DOI：
10.1016/j.cbi.2022.109978
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发表时间：
2022-05-17
影响因子：
5.1
通讯作者：
Chang, Long-Sen
中科院分区：
文献类型：
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作者：
Chiou, Jing-Ting;Lee, Yuan-Chin;Chang, Long-Sen
持续用对苯二酚（HQ）处理的人白血病U937细胞转化为MCL1和BCL2L1表达增加的U937/HQ细胞。与亲本细胞相比，U937/HQ细胞对ABT-263 （BCL2/BCL2L1抑制剂）/ABT-199 （BCL2抑制剂）的细胞毒性敏感性较低。WEHI-539 （BCL2L1抑制剂）与ABT-199或ABT-263联合使用对U937和U937/HQ细胞具有协同杀伤作用。因此，我们进一步研究了WEHI-539与ABT-199联合作用诱导细胞毒性的机制。WEHI-539和ABT-199联合处理诱导NOX4/ROS/p38 MAPK轴介导的自噬，从而加速β - trcp mRNA的转换。下调β - trcp增加Sp1表达，从而促进Sp1介导的NOXA转录，进而诱导NOXA依赖性的MCL1降解。强化表达MCL1可减轻WEHI-539 + ABT-199的细胞毒性，诱导线粒体膜电位和细胞活力丧失。单独WEHI-539诱导Sp1/NOXA轴介导的MCL1下调，而ABT-199显著降低WEHI-539的剂量，分别诱导亲本细胞和hq选择细胞中MCL1抑制约350倍和50倍。此外，WEHI-539通过诱导noxa介导的MCL1降解，使抗ABT-199的U937细胞对ABT-199细胞毒性增敏。综上所述，本研究的数据表明ABT-199和WEHI-539共同诱导了noxa依赖性的MCL1降解，MCL1的抑制主要解释了它们在亲本、红旗选择和ABT-199抗性U937细胞中的联合细胞毒性。
Human leukemia U937 cells that were continuously treated with hydroquinone (HQ) were transformed into U937/HQ cells with increased MCL1 and BCL2L1 expression. Compared with their parental cells, U937/HQ cells were less sensitive to ABT-263 (BCL2/BCL2L1 inhibitor)/ABT-199 (BCL2 inhibitor) cytotoxicity. The combination of WEHI-539 (BCL2L1 inhibitor) with either ABT-199 or ABT-263 showed synergistic cytotoxicity to U937 and U937/HQ cells. Therefore, we further investigated the cytotoxic mechanism induced by the combination of WEHI-539 and ABT-199. The combined treatment of WEHI-539 and ABT-199 induced NOX4/ROS/p38 MAPK axis-mediated autophagy, which in turn accelerated beta-TrCP mRNA turnover. Downregulation of beta-TrCP increased Sp1 expression, thereby promoting Sp1-mediated NOXA transcription, which in turn induced NOXA-dependent MCL1 degradation. Enforced expression of MCL1 alleviated the cytotoxicity of WEHI-539 plus ABT-199 to induce the loss of mitochondrial membrane potential and cell viability. WEHI-539 alone induced Sp1/NOXA axis mediated MCL1 downregulation, while ABT-199 significantly decreased the dose of WEHI-539 by approximately 350-and 50-fold to induce MCL1 suppression in parental and HQ-selected cells, respectively. Furthermore, WEHI-539 sensitized ABT-199-resistant U937 cells to ABT-199 cytotoxicity by inducing NOXA-mediated degradation of MCL1. Collectively, the data in this study indicate that ABT-199 and WEHI-539 cooperatively induce NOXA-dependent MCL1 degradation, and the inhibition of MCL1 mainly explains their combined cytotoxicity in parental, HQ-selected, and ABT-199-resistant U937 cells.