Alterations of the actin polymerization status as an apoptotic morphological effector in HL-60 cells
Alterations of the actin polymerization status as an apoptotic morphological effector in HL-60 cells
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DOI：
10.1002/(sici)1097-4644(19991215)75:4
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发表时间：
1999-12-15
影响因子：
4
通讯作者：
Hemstreet, GP
中科院分区：
文献类型：
--
作者：
Rao, JY;Jin, YS;Hemstreet, GP
细胞骨架肌动蛋白网络的改变已被牵连作为细胞凋亡的形态效应。然而，研究直接联系肌动蛋白的变化：细胞凋亡的形态学事件是缺乏的。本研究定量研究了肌动蛋白改变对喜树碱（CPT）诱导HL-60细胞凋亡过程的影响。肌动蛋白改变由两种不同类型的试剂诱导：聚合刺激剂Jasplakinase（Jas）和聚合阻断剂细胞松弛素B（CB）。肌动蛋白聚合状态的测量通过两种互补的方法：基于细胞团的DNA酶I抑制法，和基于单个细胞的定量荧光图像分析（QFIA）测定。las诱导的肌动蛋白聚合直接导致细胞凋亡。相比之下，CB，肌动蛋白聚合阻断剂，部分抑制CPT诱导的细胞凋亡。用更特异的肌动蛋白解聚剂细胞松弛素E观察到CPT诱导的凋亡反应的类似抑制。在细胞凋亡过程中，肌动蛋白聚合状态的改变分为三个连续的步骤：首先聚合，然后解聚，最后降解。然而，与CPT诱导的细胞凋亡相比，茉莉花诱导的细胞凋亡的特征在于明显的肌动蛋白聚合，对应于突出的膜起泡的形态学，但较少的凋亡小体形成。此外，DNA酶I的活性，这是通常抑制G-肌动蛋白，特别是激光处理的细胞中检测到。这些结果表明，肌动蛋白聚合的调节是一个重要的凋亡形态效应，而化学物质的肌动蛋白聚合状态的改变不仅对改变凋亡细胞的形态，但在HL-60细胞的凋亡诱导以及有深远的影响。(C)1999 Wiley-Liss，Inc.
The alterations of the cytoskeletal actin network have been implicated as a morphological effector in apoptosis. However, studies directly linking actin change: to the morphological events in apoptosis are lacking. This study quantitatively examined the effect of actin alteration on the camptothecin (CPT)-induced apoptotic process in HL-60 cells. Actin alteration was induced by two distinctive types of agent: the polymerization-stimulating agent, Jasplakinolide (Jas), and the polymerization-blocking agent, cytochalasin B (CB). The actin polymerization status was measured by two complementary methods: the cell pellet-based DNase I inhibition method, and the individual cell-based quantitative fluorescence image analysis (QFIA) assay. Actin polymerization induced by las caused apoptosis directly. By contrast, CB, an actin polymerization-blocking agent, partially inhibited CPT-induced apoptosis. A similar inhibition of the CPT-induced apoptosis response was observed with a more specific actin depolymerization agent, cytochalasin E. The alterations of the actin polymerization status occurred in three sequential steps during the apoptotic process: first polymerization, followed by depolymerization, and finally degradation. However, compared with CPT-induced apoptosis, Jas-induced apoptosis was characterized by pronounced actin polymerization that corresponded morphologically with prominent membrane blebbing, but less apoptotic body formation. Furthermore, DNase I activity, which is normally inhibited by G-actin, was specifically detected in las-treated cells. These results show that the regulation of actin polymerization is an important apoptotic morphological effector, whereas the alterations of the actin polymerization status by chemicals have profound effects not only on altering the morphology of apoptotic cells, but on apoptosis induction in HL-60 cells as well. (C) 1999 Wiley-Liss, Inc.