Document Detail


Role of caspases-3 and -7 in Apaf-1 proteolytic cleavage and degradation events during cisplatin-induced apoptosis in melanoma cells.
MedLine Citation:
PMID:  14729468     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Apoptosis protease-activating factor-1 (Apaf-1), the central element in the mitochondrial pathway of apoptosis, is frequently absent or poorly expressed in metastatic melanomas, a tumor type showing a low degree of spontaneous apoptosis and a poor response to conventional therapies. In the present study, we used the Apaf-1-positive Me665/2/21 melanoma cell line to investigate the fate of Apaf-1 during cisplatin-induced apoptosis. As novel findings described for the first time in melanoma cells, we observed that Apaf-1 was markedly decreased during apoptosis, already at early stages of cell damage; concurrently, an immunoreactive N-terminal fragment of congruent with 26 kDa was evident. In spite of the remarkable decrease of Apaf-1 in apoptotic cells, caspase-9 was found to be processed and enzymatically active. Both Apaf-1 depletion and its proteolytic cleavage were markedly prevented in presence of the caspase-3/-7 inhibitor ac-DEVD-CHO. In presence of ac-DEVD-CHO, caspase-9 activity was also inhibited, along with a partially different pattern of caspase-9 processing forms. Unexpectedly, the inhibition afforded by ac-DEVD-CHO on several components, that is, caspase-3/-7 and caspase-9 activities, and Apaf-1 proteolytic degradation, did not abrogate the apoptotic morphology and cell detachment, nor the proteolytic degradation of crucial targets, such as poly(ADP-ribose) polymerase (PARP) and lamin B. Together, our results suggest that caspase-3 and -7, proved to be dispensable for the above apoptosis-associated events, play a role on Apaf-1 handling and possibly on apoptosome function.
Authors:
Barbara Del Bello; Marta A Valentini; Paola Mangiavacchi; Mario Comporti; Emilia Maellaro
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Experimental cell research     Volume:  293     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  2004 Feb 
Date Detail:
Created Date:  2004-01-19     Completed Date:  2004-03-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  302-10     Citation Subset:  IM    
Affiliation:
Department of Pathophysiology, Experimental Medicine and Public Health, University of Siena, via A. Moro, 53100 Siena, Italy.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis / drug effects*,  physiology
Apoptotic Protease-Activating Factor 1
Caspase 3
Caspase 7
Caspases / antagonists & inhibitors,  metabolism*
Cell Adhesion / drug effects,  physiology
Cell Line, Tumor
Cisplatin / pharmacology*
Cytochromes c / metabolism
Down-Regulation / drug effects,  physiology
Drug Resistance, Neoplasm / physiology
Enzyme Inhibitors / pharmacology
Humans
Lamin Type B / metabolism
Macromolecular Substances
Melanoma / drug therapy,  enzymology*
Mitochondria / drug effects,  enzymology
Peptide Fragments / metabolism
Peptide Hydrolases / drug effects,  metabolism
Poly(ADP-ribose) Polymerases
Proteins / drug effects,  metabolism*
Signal Transduction / drug effects,  physiology
Chemical
Reg. No./Substance:
0/APAF1 protein, human; 0/Apoptotic Protease-Activating Factor 1; 0/Enzyme Inhibitors; 0/Lamin Type B; 0/Macromolecular Substances; 0/Peptide Fragments; 0/Proteins; 15663-27-1/Cisplatin; 9007-43-6/Cytochromes c; EC 2.4.2.30/PARP1 protein, human; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.-/Peptide Hydrolases; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/CASP7 protein, human; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 7; EC 3.4.22.-/Caspases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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