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Titolo:
Stathmo-apoptosis: Arresting apoptosis by fluorochrome-labeled inhibitor of caspases
Autore:
Smolewski, P; Grabarek, J; Phelps, DJ; Darzynkiewicz, Z;
Indirizzi:
New York Med Coll, Brander Canc Res Inst, Valhalla, NY 10595 USA New York Med Coll Valhalla NY USA 10595 Res Inst, Valhalla, NY 10595 USA Med Univ Lodz, Dept Hematol, Lodz, Poland Med Univ Lodz Lodz PolandMed Univ Lodz, Dept Hematol, Lodz, Poland Pomerian Sch Med, Dept Pathol, Szczecin, Poland Pomerian Sch Med Szczecin Poland Sch Med, Dept Pathol, Szczecin, Poland Intergen Co, Purchase, NY USA Intergen Co Purchase NY USAIntergen Co, Purchase, NY USA
Titolo Testata:
INTERNATIONAL JOURNAL OF ONCOLOGY
fascicolo: 4, volume: 19, anno: 2001,
pagine: 657 - 663
SICI:
1019-6439(200110)19:4<657:SAABFI>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
MCF-7 CELLS; CAMPTOTHECIN; DEATH; PHASE;
Keywords:
stathmo-kinesis; apoptosis; caspases; inhibitors; cell death rate; cell necrobiology;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
18
Recensione:
Indirizzi per estratti:
Indirizzo: Darzynkiewicz, Z NYMC, Brander Canc Res Inst, 19 Bradhurst Ave,Suite 2400,Hawthorne, NY 10532 USA NYMC 19 Bradhurst Ave,Suite 2400 Hawthorne NY USA 10532 A
Citazione:
P. Smolewski et al., "Stathmo-apoptosis: Arresting apoptosis by fluorochrome-labeled inhibitor of caspases", INT J ONCOL, 19(4), 2001, pp. 657-663

Abstract

Apoptosis, like mitosis, is a kinetic event. The entire duration of apoptosis, from its onset to total disintegration of the cell, is often short andmay be of variable duration. The time-window through which individual apoptotic cells display their characteristic features that serve to identify them varies depending on: a) the assay that is used, b) the cell type, c) thenature of the inducer of apoptosis, and d) the environmental factors the cell is exposed to that may shorten or prolong apoptosis. Thus, because the apoptotic index (AI) does not accurately represent incidence of apoptosis it is desirable to estimate the rate of cell death in analogy to the cell birth rate which is assessed by the stathmo-kinetic approach by arresting cells in mitosis. In this study the fluorescent caspase inhibitor FAM-VAD-FMK was used for dual purposes: a) to arrest the process of apoptosis (stathmo-apoptosis), and b) to have the arrested cells labeled with fluorochrome. Apoptosis of HL-60 and MCF-7 cells was induced by DNA topoisomerase I inhibitor camptothecin (CPT) and FAM-VAD-FMK was added at the same time as the inducer. While the cells become progressively labeled with FAM-VAD-FMK, their disintegration, loss of the phase-contrast and loss of the capability to bind the inhibitor, and in the case of MCF-7 cells, detachment from the slides, all were prevented for up to 48 h. The percentage of FAM-VAD-FMK labeledHL-60 cells was plotted as a function of time after addition of CPT and the rate of cell entrance to apoptosis was estimated from the slopes of the stathmo-apoptotic plot at different time after administration of CPT. The plot revealed the presence of two distinct subpopulations: during the initial8 h of the treatment with CPT the cells of the first subpopulation, predominantly the S-phase cells, were entering apoptosis at a rate of about 7% ofcells per hour. The remaining cells were stochastically entering apoptosisbetween 8 and 48 h at a rate 1% of cells per hour. The present approach offers a unique capability to accurately estimate the kinetics of cell transition to apoptosis, revealing the unbiased cumulative apoptotic index over along time span after induction of apoptosis.

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Documento generato il 30/11/20 alle ore 10:07:04