Kinetics of Smac/DIABLO release from mitochondria during apoptosis of MCF-7 breast cancer cells

被引:21
作者
Gorka, M
Godlewski, MM
Gajkowska, B
Wojewodzka, U
Motyl, T
机构
[1] Agr Univ Warsaw, Fac Vet Med, Dept Physiol Sci, PL-02776 Warsaw, Poland
[2] Polish Acad Sci, Med Res Ctr, Lab Cell Ultrastruct, Warsaw, Poland
关键词
Smac/DIABLO; Bax; apoptosis; homeostatic confocal microscopy;
D O I
10.1016/j.cellbi.2004.07.003
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Smac/DIABLO, a pro-apoptotic protein released from mitochondrial intermembrane space during apoptosis, promotes caspase activation by IAPs neutralization. The kinetics and molecular mechanism of Smac/DIABLO release from mitochondria has remained obscure. Homeostatic confocal microscopy, for the first time, showed the precise kinetics of Smac/DIABLO release from mitochondria during CPT-induced apoptosis in living MCF-7 cells. The time pattern of Smac/DIABLO escape from mitochondria comprised two phases: the initial phase of gradual protein release. followed by the second phase of plateau, appearing, after 24 min of cell exposure to the drug. A similar pattern was observed during oxidative stress. The dynamics of Smac/DIABLO redistribution was confirmed by different methods: traditional confocal microscopy, immunoelectron microscopy and laser scanning cytometry. The inhibition of m-calpain prevented Smac/DIABLO release from mitochondria. which confirmed the involvement of Bax in the process. Acquired results indicate that CPT treatment triggers Bax-dependent release of Smac/DIABLO from mitochondria simultaneously with the efflux of cytochrome c. (C) 2004 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:741 / 754
页数:14
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