Document Detail


Assessment of apoptosis by immunohistochemistry to active caspase-3, active caspase-7, or cleaved PARP in monolayer cells and spheroid and subcutaneous xenografts of human carcinoma.
MedLine Citation:
PMID:  19029405     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Immunohistochemistry to active caspase-3, recently recommended for apoptosis detection, is inappropriate to detect apoptosis involving caspase-7. Cleavage of poly-ADP-ribose polymerase 1 (PARP-1), a major substrate of both caspases, is a valuable marker of apoptosis. Apoptosis evaluation induced in vitro either by paclitaxel or by photodynamic treatment (PDT) with Foscan in HT29 or KB monolayer cells and HT29 spheroids yielded a close percentage of labeled cells whatever the antibody used, whereas in control specimens, cleaved PARP (c-PARP) immunostaining failed to detect apoptosis as efficiently as active caspase-3 or -7 immunostaining. Studies in MDA-MB231 monolayer cells and HT29 xenografts either subjected or not subjected to Foscan-PDT resulted in a significant higher number of active caspase-3-labeled cells, although immunofluorescence analysis showed c-PARP and active caspase-3 perfectly colocalized in tumors. A restricted expression of c-PARP was obvious in the greater part of caspase-3 expressing cells from control tumor, whereas photosensitized tumors showed a higher number of cells expressing large fluorescent spots from both active caspase-3 and c-PARP. These results support the assumption that c-PARP expression was dependent on treatment-induced apoptosis. The absence of caspase-7 activation in some caspase-3-expressing cells undergoing Foscan-PDT shows the relevance of using antibodies that can discriminate caspase-dependent apoptotic pathways.
Authors:
Aude Bressenot; Sophie Marchal; Lina Bezdetnaya; Julie Garrier; François Guillemin; François Plénat
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Publication Detail:
Type:  Journal Article     Date:  2008-11-24
Journal Detail:
Title:  The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society     Volume:  57     ISSN:  0022-1554     ISO Abbreviation:  J. Histochem. Cytochem.     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-03-17     Completed Date:  2009-04-08     Revised Date:  2010-09-23    
Medline Journal Info:
Nlm Unique ID:  9815334     Medline TA:  J Histochem Cytochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  289-300     Citation Subset:  IM    
Affiliation:
Service d'Anatomie et de Cytologie Pathologiques, Hôpital de Brabois, Centre Hospitalier Régional et Universitaire Nancy, Vandoeuvre-lès-Nancy, France.
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MeSH Terms
Descriptor/Qualifier:
Adenocarcinoma / metabolism,  pathology*
Animals
Antineoplastic Agents / pharmacology
Apoptosis*
Caspase 3 / metabolism*
Caspase 7 / metabolism*
Colonic Neoplasms / metabolism,  pathology*
Humans
Immunohistochemistry
Mesoporphyrins / pharmacology
Mice
Mice, Nude
Neoplasm Transplantation
Paclitaxel / pharmacology
Poly(ADP-ribose) Polymerases / metabolism*
Spheroids, Cellular / metabolism*
Transplantation, Heterologous
Triazenes / pharmacology
Chemical
Reg. No./Substance:
0/Antineoplastic Agents; 0/Mesoporphyrins; 0/Triazenes; 0/temoporfin; 33069-62-4/Paclitaxel; 7227-91-0/1-phenyl-3,3-dimethyltriazene; EC 2.4.2.30/PARP1 protein, human; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 7
Comments/Corrections

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


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