BAX and BAK proteins are required for cyclin-dependent kinase inhibitory drugs to cause apoptosis
Identificadores
URI: http://hdl.handle.net/20.500.12020/1186ISSN: 1538-8514
DOI: 10.1158/1535-7163.MCT-08-0655
Autor/es
Garrofé-Ochoa, Xènia; Melero Fernandez de Mera, Raquel Maria; Fernandez Gomez, Francisco Jose; Ribas, Judit; Jordan, Joaquin; [et al.]Fecha
2008-12-03Tipo de documento
articleÁrea/s de conocimiento
Ciencias BiomédicasResumen
In previous reports, we have shown in SH-SY5 cells that olomoucine and roscovitine, two inhibitory drugs of cyclin-dependent kinases, caused apoptosis independent of the extrinsic pathway. In this experimental paradigm, apoptosis was refractory to the protective effects of either Bcl-2 or Bcl-XL overexpression. We are now reporting that the failure of Bcl-XL to prevent dell death was consistent with no effect on the kinetics of caspase activation and cytochrome c release. To further characterize this issue, we have discarded a direct effect of either olomoucine or roscovitine on mitochondrial permeability transition. Moreover, we have evidence that an intrinsic pathway took place in SH-SY5Y cells by showing the mitochondrial translocation of a GFP-Bax construct on transfection and treatment with cyclin-dependent kinase inhibitory drugs. Finally, we tested the effect of olomoucine and roscovitine on wild-type, bax-/-, bak-/-, and double bax-/-bak -/- mouse embryonic fibroblasts (MEF). In wild-type MEFs, both drugs induced cell death by apoptosis in a dose-dependent manner. In bax -/-, bak-/-, and, particularly, double bax -/-bak-/- MEFs, we observed the inhibition of apoptosis. In conclusion, olomoucine and roscovitine caused apoptosis through an intrinsic pathway, with Bax and Bak proteins being involved.