Maruyama W, Irie S, Sato T A
Laboratory of Molecular Oncology, Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
Histochem J. 2000 Aug;32(8):495-503. doi: 10.1023/a:1004104619154.
To investigate the early event of apoptosis, we monitored the morphological changes in the early stage of Fas-induced apoptosis in the human T-cell lymphoma cell line Jurkat, using confocal microscopy. Morphological changes in the nuclei were observed from 30 min after stimulation, and preceded the changes in the cytoskeleton. This kind of change was enhanced in the presence of EGTA but decreased in the presence of dihydrocytochalasin B, without any changes in caspase-3 activation. During the changes in shape of the cells, the actin cytoskeleton collapsed and shrank in the center. Even though nuclei also changed their shapes in apoptotic cells, they were partially TUNEL-negative, suggesting that they were not yet damaged at the DNA level. Our results suggest that, in the process of apoptosis in Jurkat cells, cell nuclei and cytoskeleton are changed first, then membrane blebbing and caspase-3 activation occur, and fragmentation of chromosomal DNA is last.
为了研究凋亡的早期事件,我们使用共聚焦显微镜监测了人T细胞淋巴瘤细胞系Jurkat中Fas诱导凋亡早期的形态变化。刺激后30分钟观察到细胞核的形态变化,且早于细胞骨架的变化。在存在乙二醇双四乙酸(EGTA)的情况下这种变化增强,而在存在二氢细胞松弛素B的情况下则减弱,同时半胱天冬酶-3的激活没有任何变化。在细胞形状变化过程中,肌动蛋白细胞骨架在中心塌陷并收缩。尽管凋亡细胞中的细胞核也改变了形状,但它们部分为TUNEL阴性,这表明它们在DNA水平上尚未受损。我们的结果表明,在Jurkat细胞凋亡过程中,细胞核和细胞骨架首先发生变化,然后出现膜泡化和半胱天冬酶-3激活,最后是染色体DNA片段化。