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Vol. 61, Issue 1, 142-149, January 2002
The Pediatric Center for Neuroscience, Children's Hospital of
Pittsburgh, Pittsburgh, Pennsylvania
Previous studies from our laboratory have demonstrated that Bcl-2 has a
proapoptotic effect on neocarzinostatin (NCS)-treated PC12
pheochromocytoma cells. In the present study, we examine the mechanisms
of this effect and demonstrate its relevance for the in vivo situation.
Four hours after NCS treatment, a 23-kDa cleavage product of Bcl-2 was
detected in whole cell lysates of bcl-2-transfected
PC12 cells. In contrast, bcl-2 transfection protected
PC12 cells from cisplatin-induced apoptosis, and cisplatin treatment
did not result in Bcl-2 cleavage. Similarly, Bcl-2 cleavage did not
occur and Bcl-2-mediated protection from, rather than potentiation of
apoptosis was observed after NCS treatment of MCF-7 breast cancer
cells. The caspase 3-specific inhibitor Ac-DEVD-CHO prevented Bcl-2
cleavage and attenuated NCS-induced apoptosis in
bcl-2-transfected PC12 cells, whereas it had no effect
on NCS-induced apoptosis in mock-transfected PC12 cells. Furthermore,
MCF-7 cells do not express caspase 3, a finding in concert with the
lack of Bcl-2 cleavage in this line. In in vivo experiments, xenografts of bcl-2-transfected PC12 cells were more susceptible
to NCS toxicity than were xenografts of mock-transfected PC12 cells.
Caspase 3-mediated Bcl-2 cleavage therefore plays an important role in
the potentiation by Bcl-2 of NCS-induced apoptosis.
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