BIMEL is a key effector molecule in oxidative stress-mediated apoptosis in acute myeloid leukemia cells when combined with arsenic trioxide and buthionine sulfoximine.
BIMEL is a key effector molecule in oxidative stress-mediated apoptosis in acute myeloid leukemia cells when combined with arsenic trioxide and buthionine sulfoximine.
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DOI：
10.1186/1471-2407-14-27
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发表时间：
2014-01-15
期刊：
影响因子：
3.8
通讯作者：
Fujii Y
中科院分区：
文献类型：
--
作者：
Tanaka Y;Komatsu T;Shigemi H;Yamauchi T;Fujii Y
三氧化二砷（ATO）通过诱导细胞凋亡而成为治疗急性早幼粒细胞白血病（APL）的有效药物。氧化应激途径调节剂丁硫丙啶亚砜亚胺（BSO）被认为是治疗ATO不敏感性白血病的潜在联合用药。然而，BSO介导的ATO诱导的细胞凋亡增强的确切机制尚未完全了解。本研究比较了ATO/BSO和ATO单独作用于HL 60白血病细胞后细胞死亡的差异，并探讨了BSO增强ATO诱导的细胞死亡的分子机制。HL 60 APL细胞用于研究。用免疫沉淀和免疫印迹分析一系列信号分子的激活和表达。凋亡细胞死亡检测半胱天冬酶和聚（ADP-核糖）聚合酶激活。使用氧化还原敏感染料测定细胞内活性氧（ROS）的产生。线粒体外膜透化观察与共聚焦显微镜使用近红外染料和细胞色素c的释放与免疫印迹法测定。小干扰（si）RNA用于抑制基因表达。在BSO存在下，HL 60细胞对ATO变得更敏感。ATO/BSO诱导的线粒体损伤伴随着线粒体外膜透性降低、细胞色素c释放和caspase激活。抗氧化剂可抑制ATO/BSO诱导的线粒体损伤。在处理的细胞中，添加BSO诱导促凋亡BCL 2蛋白BIMEL和抗凋亡BCL 2蛋白MCL 1的磷酸化。磷酸化的BIMEL从MCL 1上解离下来，与BAX相互作用，随后BAX发生构象变化。此外，用小干扰RNA敲低BIMEL抑制BSO对ATO诱导的凋亡的增强。BSO对ATO诱导的细胞死亡的增强作用在分子水平上进行了表征，以供临床使用。加入BSO诱导线粒体损伤介导的凋亡通过磷酸化的BIMEL和MCL 1，导致其解离，并增加BIMEL和BAX之间的相互作用。
Arsenic trioxide (ATO) is reported to be an effective therapeutic agent in acute promyelocytic leukemia (APL) through inducing apoptotic cell death. Buthionine sulfoximine (BSO), an oxidative stress pathway modulator, is suggested as a potential combination therapy for ATO-insensitive leukemia. However, the precise mechanism of BSO-mediated augmentation of ATO-induced apoptosis is not fully understood. In this study we compared the difference in cell death of HL60 leukemia cells treated with ATO/BSO and ATO alone, and investigated the detailed molecular mechanism of BSO-mediated augmentation of ATO-induced cell death. HL60 APL cells were used for the study. The activation and expression of a series of signal molecules were analyzed with immunoprecipitation and immunoblotting. Apoptotic cell death was detected with caspases and poly (ADP-ribose) polymerase activation. Generation of intracellular reactive oxygen species (ROS) was determined using a redox-sensitive dye. Mitochondrial outer membrane permeabilization was observed with a confocal microscopy using NIR dye and cytochrome c release was determined with immunoblotting. Small interfering (si) RNA was used for inhibition of gene expression. HL60 cells became more susceptible to ATO in the presence of BSO. ATO/BSO-induced mitochondrial injury was accompanied by reduced mitochondrial outer membrane permeabilization, cytochrome c release and caspase activation. ATO/BSO-induced mitochondrial injury was inhibited by antioxidants. Addition of BSO induced phosphorylation of the pro-apoptotic BCL2 protein, BIMEL, and anti-apoptotic BCL2 protein, MCL1, in treated cells. Phosphorylated BIMEL was dissociated from MCL1 and interacted with BAX, followed by conformational change of BAX. Furthermore, the knockdown of BIMEL with small interfering RNA inhibited the augmentation of ATO-induced apoptosis by BSO. The enhancing effect of BSO on ATO-induced cell death was characterized at the molecular level for clinical use. Addition of BSO induced mitochondrial injury-mediated apoptosis via the phosphorylation of BIMEL and MCL1, resulting in their dissociation and increased the interaction between BIMEL and BAX.
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影响因子：
16.2
作者：
Putcha, GV;Le, SY;Johnson, EM
通讯作者：
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影响因子：
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