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.

Abstract

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
• Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (J Histochem Cytochem) Vol. 57 Issue 4 Pg. 289-300 (Apr 2009) ISSN: 0022-1554 [Print] United States
• PMID: 19029405 (Publication Type: Journal Article)

Chemical References

• Antineoplastic Agents
• Mesoporphyrins
• Triazenes
• 1-phenyl-3,3-dimethyltriazene
• PARP1 protein, human
• Poly (ADP-Ribose) Polymerase-1
• Poly(ADP-ribose) Polymerases
• Caspase 3
• Caspase 7
• temoporfin
• Paclitaxel

Topics

• 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) Polymerase-1
• Poly(ADP-ribose) Polymerases (metabolism)
• Spheroids, Cellular (metabolism)
• Transplantation, Heterologous
• Triazenes (pharmacology)