伊马替尼通过激活Caspase降解慢性髓细胞白血病细胞中的锌指蛋白ZFX
首发时间:2018-12-21
摘要:目的:检测伊马替尼(IM)处理对慢性髓细胞白血病(CML)细胞中锌指蛋白ZFX表达的影响并初探其机制。方法:Q-RT-PCR和western blot分析IM处理后K562细胞中ZFX的mRNA和蛋白表达;免疫荧光法检测IM处理后CML患者CD34+细胞中ZFX蛋白的表达;钙蛋白酶活性试剂盒检测IM处理后K562细胞内钙蛋白酶的活性;IM处理对照和自噬相关基因ATG7敲除的K562细胞,而后检测ZFX蛋白表达;比较经IM单独和联合蛋白酶体抑制剂MG132处理的K562细胞中ZFX的蛋白表达;检测经IM单独或联合泛Caspase抑制剂Z-VAD-FMK处理后的K562细胞的凋亡、Caspase活化状态和ZFX蛋白的表达。结果:IM处理K562细胞后,ZFX的mRNA水平不变,但蛋白表达降低;IM处理CML患者CD34+细胞后,ZFX表达呈剂量依赖性降低;IM处理细胞的钙蛋白酶活性不变;干扰自噬或抑制蛋白酶体功能不影响IM对ZFX蛋白表达的影响;IM诱导K562细胞发生凋亡并伴随Caspase活性增强,Z-VAD-FMK处理抑制上述效应并使得ZFX蛋白表达有所恢复。结论:IM通过诱发CML细胞中Caspase活化降解锌指蛋白ZFX。
关键词: 伊马替尼 锌指蛋白ZFX 细胞凋亡 Caspase 慢性髓细胞白血病
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Imatinib mesylate activates Caspase to degrade zinc finger protein X-linked in chronic myeloid leukemia cells
Abstract:Objective: To study the effect of imatinib mesylate (IM) on the expression of zinc finger protein X-linked (ZFX) in chronic myeloid leukemia (CML) cells and its underlying mechanism. Methods:Q-RT-PCR and western blot were used to analyze the expression of ZFX in K562 cells with and without IM treatment; immunofluorescence was used to analyze ZFX expression upon IM treatment in CD34+ cells from CML patients;calpain activity was compared between cells treated with and without IM; ZFX protein expression was analyzed in cells treated only with IM and those treated with IM and MG132, a proteasome inhibitor; ZFX protein expression was compared between K562 and ATG7-null K562 cells upon IM treatment; cell apoptosis, caspase activation and ZFX expression were analyzed in K562 cells treated with IM and those combined with Z-VAD-FMK (a caspase inhibitor). Results: IM treatment caused adose-dependent decrease of ZFX expression on protein level rather than on mRNA level in K562 cells.IM treatment led to dose-dependent decrease of ZFX in CML CD34+ cells. IM treatment didn\'t change calpain activity of K562 cells, impaired proteasome or autophagy didn\'t "rescue" the decrease of ZFX protein upon IM treatment. IM treatment induced apoptosis of K562 cells, while additional Z-VAD-FMK treatment inhibited the cellular apoptosis and "rescued" ZFX protein expression. Conclusion: Imatinib induced apoptosis of CML cells and activation of caspase, which was responsible for the degradation of ZFX protein.
Keywords: Imatinib ZFX Apoptosis Caspase Chronic myeloid leukemia
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伊马替尼通过激活Caspase降解慢性髓细胞白血病细胞中的锌指蛋白ZFX
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