Induction of Apoptosis in Human Leukemia Cells by the CDK1 Inhibitor CGP74514A
Induction of Apoptosis in Human Leukemia Cells by the CDK1 Inhibitor CGP74514A
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DOI：
10.4161/cc.1.2.116
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发表时间：
2002-03-01
期刊：
影响因子：
4.3
通讯作者：
Grant, Steven
中科院分区：
文献类型：
--
作者：
Dai, Yun;Dent, Paul;Grant, Steven
我们研究了CDK 1抑制剂CGP 74514 A对人白血病细胞中细胞周期和凋亡相关事件的影响。暴露于5 μ MCGP 74514 A 18小时诱导线粒体损伤（即，Δ psi（m）的损失）和多种人白血病细胞系中的细胞凋亡（例如，U937、HL-60、KG-1、CCRF-CEM、Raji和THP;范围30-95%）。在U937细胞中，CGP 74514 A诱导的细胞凋亡（5 μ M）在4小时内变得明显，并接近100%的24小时。泛半胱天冬酶抑制剂Boc-fastin和半胱天冬酶-8抑制剂IETD-fastin反对CGP 74514 A诱导的半胱天冬酶-9激活和PARP降解，但不是细胞色素c或Smac/DIABLO释放。CGP 74514 A介导的凋亡基本上被全长Bcl-2、环缺失突变体Bcl-2和Bcl-xL的异位表达阻断。CGP 74514 A处理（5 μ M; 18小时）导致p21 CIP 1表达增加、p27 KIP 1降解、E2 F1表达减少和p34 cdc 2去磷酸化。它还诱导早期（即，在2小时内）抑制CDK 1活性和pRb的去磷酸化，随后是pRb降解，但不阻断在CDK 2和CDK 4特异性位点的pRb磷酸化。这些发现表明，选择性CDK 1抑制剂，CGP 74514 A，诱导复杂的变化，在人类白血病细胞的细胞周期相关蛋白伴随着广泛的线粒体损伤，半胱天冬酶激活，和凋亡。
We have examined the effects of the CDK1 inhibitor CGP74514A on cell cycle-and apoptosis-related events in human leukemia cells. An 18-hr exposure to 5 mu M CGP74514A induced mitochondrial damage (i.e., loss of Delta psi(m)) and apoptosis in multiple human leukemia cell lines (e.g., U937, HL-60, KG-1, CCRF-CEM, Raji, and THP; range 30-95%). In U937 cells, CGP74514A-induced apoptosis (5 mu M) became apparent within 4 hr and approached 100% by 24 hr. The pan-caspase inhibitor Boc-fmk and the caspase-8 inhibitor IETD-fmk opposed CGP74514A-induced caspase-9 activation and PARP degradation, but not cytochrome c or Smac/DIABLO release. CGP74514A-mediated apoptosis was substantially blocked by ectopic expression of full-length Bcl-2, a loop-deleted mutant Bcl-2, and Bcl-xL. CGP74514A treatment (5 mu M; 18 hr) resulted in increased p21CIP1 expression, p27KIP1 degradation, diminished E2F1 expression, and dephosphorylation of p34cdc2. It also induced early (i.e., within 2 hr) inhibition of CDK1 activity and dephosphorylation of pRb, followed by pRb degradation, but did not block pRb phosphorylation at CDK2- and CDK4-specific sites. These findings indicate that the selective CDK1 inhibitor, CGP74514A, induces complex changes in cell cycle-related proteins in human leukemia cells accompanied by extensive mitochondrial damage, caspase activation, and apoptosis.