Early detection of staurosporine-induced apoptosis by comet and annexin V assays |
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Authors: | Thierry Godard Edwige Deslandes Pierre Lebailly Carole Vigreux François Sichel Jean-Michel Poul Pascal Gauduchon |
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Institution: | GRECAN and INSERM CJF 96–03, Université de Caen, F-14032 Caen Cedex, France, FR AFSSA (Agence Fran?aise de Sécurité Sanitaire des Aliments), Unité de Toxicologie, Javené, F-35133 Fougeres, France, FR
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Abstract: | Comet, TUNEL, and annexin V assays were used to identify DNA fragmentation and plasma membrane alterations occurring during
staurosporine-induced apoptosis in Chinese hamster ovary cells. TUNEL assay detected apoptotic cells after 6 h treatment.
The occurrence of annexin V immunofluorescence staining after 1 h treatment confirms that exposure of phosphatidylserine (PS)
residues is an early biochemical feature of apoptosis. According to intensity, three annexin staining patterns were distinguished,
related to different steps in the apoptotic process. The detection of highly damaged cells by the comet assay after 3 h treatment
occurred earlier than the detection of DNA modifications by the TUNEL assay, but later than the exposure of PS residues. However,
late apoptotic cells, otherwise characterized by plasma membrane disruption and high annexin V staining, were not detected
by the comet assay. In this case, comet assay modified by omitting electrophoresis (halo assay) was more sensitive for an
accurate quantification of the apoptotic fraction.
Accepted: 2 June 1999 |
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