Comparative effects of Bcl-2 over-expression and ZVAD.FMK treatment on dexamethasone and VP16-induced apoptosis in CEM cells.
It is becoming apparent that caspases are essential mediators of the execution phase of apoptosis. A decrease in mitochondrial membrane potential (PsiM) is also thought to be an early event in apoptosis. In this study, we compare the effects of Bcl-2 over-expression against N-benzyloxycabonyl-Val-Ala-Asp-fluoromethylketone (ZVAD. FMK)-sensitive caspase blockade on dexamethasone (DEX) and etoposide (VP16)-induced apoptosis in CEM T lymphoid cells. We assessed changes in nuclear chromatin, cell size, fragmentation, cell membrane permeability and PsiM. We found Bcl-2 over-expression and ZVAD.FMK-sensitive caspase inhibition were able to prevent chromatin condensation and cellular fragmentation. However, ZVAD.FMK was neither able to prevent loss of plasma membrane integrity nor PsiM depolarization which occur in both VP16 and DEX-induced apoptosis. In VP16-induced apoptosis, the increase in cell membrane permeability was actually potentiated by caspase inhibition. Interestingly, ZVAD.FMK did prevent VP16-induced but not DEX-induced cell shrinkage. These results suggest that not all the actions of Bcl-2 can be explained by its ability to prevent caspase activation. Rather Bcl-2 must have other targets of action which include functions associated with mitochondria.
Benson
R S
RS
School of Biological Sciences, The University of Manchester, Oxford Road, Manchester, UK.
Dive
C
C
Watson
A J
AJ
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D061945:Caspase Inhibitors; D002463:Cell Membrane Permeability / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002843:Chromatin / Q000378:metabolism; D003907:Dexamethasone; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005047:Etoposide; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D013601:T-Lymphocytes / Q000187:drug effects*; D014407:Tumor Cells, Cultured
D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D002843:Chromatin; D004791:Enzyme Inhibitors; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D005047:Etoposide; D003907:Dexamethasone