Nα-Tosyl-L-phenylalanine Chloromethyl Ketone Induces Caspase-dependent Apoptosis in Transformed Human B Cell Lines with Transcriptional Down-regulation of Anti-apoptotic HS1-associated Protein X-1
Nα-Tosyl-L-phenylalanine Chloromethyl Ketone Induces Caspase-dependent Apoptosis in Transformed Human B Cell Lines with Transcriptional Down-regulation of Anti-apoptotic HS1-associated Protein X-1
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DOI：
10.1074/jbc.m109.027912
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发表时间：
2009-10-09
影响因子：
4.8
通讯作者：
Fadeel, Bengt
中科院分区：
文献类型：
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作者：
Jitkaew, Siriporn;Trebinska, Alicja;Fadeel, Bengt
N-α-ToSyl-L-苯丙氨酸氯甲基酮（TPCK）已被广泛用于研究参与基因表达和细胞存活/细胞死亡的信号转导途径。然而，TPCK对细胞凋亡的相互矛盾的影响已被报道，导致TPCK诱导的促进或预防细胞凋亡的潜在信号事件尚未完全理解。在这里，我们表明，TPCK诱导半胱天冬酶依赖性细胞凋亡的EB病毒（EBV）转化的人B细胞系与释放促凋亡蛋白从线粒体。TPCK处理还导致抗凋亡蛋白cIAP 1、cIAP 2和HAX-1的下调，以及抗凋亡蛋白Bcl-2和XIAP的半胱天冬酶依赖性切割。定量PCR分析证实TPCK诱导的HAX-1下调发生在转录水平，并且使用特异性药理学抑制剂Bay 11-7082的实验表明HAX-1表达受转录因子的调节，来自HAX 1纯合突变患者的NF-κ B细胞系对TPCK-1更敏感。诱导凋亡，与正常供体细胞系相比。此外，N-乙酰半胱氨酸有效地阻断TPCK诱导的EBV转化的B细胞系的凋亡，并阻止抗凋亡蛋白的下调或切割。总之，我们的研究表明，TPCK诱导人B细胞系的凋亡，并发挥多种作用的促凋亡和抗凋亡因子。
N-alpha-ToSyl-L-phenylalanine chloromethylketone (TPCK) has been widely used to investigate signal transduction pathways that are involved in gene expression and cell survival/cell death. However, contradictory effects of TPCK on apoptosis have been reported, and the underlying signaling events leading to TPCK-induced promotion or prevention of apoptosis are not fully understood. Here, we show that TPCK induces caspase-dependent apoptosis in Epstein-Barr virus (EBV)-transformed human B cell lines with release of pro-apoptotic proteins from mitochondria. TPCK treatment also results in down-regulation of the anti-apoptotic proteins, cIAP1, cIAP2, and HAX-1, and caspase-dependent cleavage of the anti-apoptotic proteins, Bcl-2 and XIAP. Quantitative PCR analysis confirmed that the TPCK-induced down-regulation of HAX-1 occurred at the transcriptional level, and experiments using the specific pharmacological inhibitor, Bay 11-7082, suggested that HAX-1 expression is subject to regulation by the transcription factor, NF-kappa B cell lines derived from patients with homozygous HAX1 mutations were more sensitive to TPCK-induced apoptosis when compared with normal donor cell lines. Furthermore, N-acetylcysteine effectively blocked TPCK-induced apoptosis in EBV-transformed B cell lines and prevented the down-regulation or cleavage of anti-apoptotic proteins. Taken together, our studies demonstrate that TPCK induces apoptosis in human B cell lines and exerts multiple effects on pro- and anti-apoptotic factors.