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Involvement of CED-3/ICE proteases in the apoptosis of B-chronic
lymphocytic leukemia cells
B Bellosillo, M Dalmau, D Colomer and J Gil
Departament de Ciencies Fisiologiques Humanes i de la Nutricio, Universitat
de Barcelona, Spain.
B-chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation
of long-lived B lymphocytes that express high levels of Bcl-2. We examined
the involvement of CED-3/ICE-like proteases in the apoptosis of B-CLL
cells. One of the substrates of these proteases is poly(ADP [adenosine
5'-diphosphate]-ribose) polymerase (PARP). The effect of different factors
that induce the apoptosis of B-CLL cells on the proteolytic cleavage of
PARP has been studied. Treatment of B-CLL cells with different
concentrations of dexamethasone (1 to 1,000 micromol/L) induced in a
dose-dependent manner the cleavage of PARP. Dexamethasone induced PARP
cleavage after 12 hours of incubation, which was almost complete at 48
hours. PARP cleavage during apoptosis of B- CLL cells was studied in cells
from eight patients and a correlation was found between cell viability and
the degree of PARP cleavage. Incubation in vitro of B-CLL cells with
fludarabine for 48 hours induced PARP cleavage in all the cases studied.
Protein kinase C (PKC) activation with 100 nmol/L TPA
(12-O-tetradecanoylphorbol 13-acetate) or incubation with interleukin-4 (10
ng/mL) prevented either dexamethasone- or fludarabine-induced proteolysis
of PARP. Incubation of B-CLL cells with the CED-3/ICE-like protease
inhibitor Z-VAD.fmk inhibited spontaneous and dexamethasone-induced PARP
cleavage and DNA fragmentation in a dose-dependent manner. Furthermore,
Z-VAD.fmk prevented the cytotoxic effect of dexamethasone. These results
indicate that CED-3/ICE-like proteases play an important role in the
apoptosis of B-CLL cells.
Volume 89,
Issue 9,
pp. 3378-3384,
05/01/1997
Copyright © 1997 by The American Society of Hematology
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