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Am J Physiol Lung Cell Mol Physiol (February 7, 2003). doi:10.1152/ajplung.00172.2002
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Submitted on June 4, 2002
Accepted on February 6, 2003

Ceramide accumulation precedes caspase-3 activation during apoptosis of A549 human lung adenocarcinoma cells

Tommer Ravid1, Adili Tsaba1, Peter Gee1, Reuven Rasooly1, Edward A. Medina1, and Tzipora Goldkorn1*

1 Department of Internal Medicine, University of California, Davis, Davis, California, USA

* To whom correspondence should be addressed. E-mail: ttgoldkorn{at}ucdavis.edu.

Ceramide, the basic structural unit of sphingolipids, controls the balance between cell growth and death by inducing apoptosis. We have previously shown that accumulation of ceramide, triggered by hydrogen peroxide (H2O2) or by short-chain ceramide analogs, induces apoptosis of lung epithelial cells. Here we elucidate the link between caspase-3 activation, at the execution phase, and ceramide accumulation, at the commitment phase of apoptosis in A549 human lung adenocarcinoma cells. The induction of ceramide accumulation by various triggers of ceramide generation such as H2O2, C6-ceramide or UDP-glucose-ceramide glucosyltransferase inhibitor, PDMP, triggered the activation of caspase-3. This ceramide elevation also induced the cleavage of the death substrate poly (ADP-ribose) polymerase and was followed by apoptotic cell death. Ceramide-mediated apoptosis was blocked by a general caspase inhibitor Boc-D-FMK, and by overexpression of the anti-apoptotic protein Bcl-2. Notably, overexpression of Bcl-2 reduced the basal cellular levels of ceramide and prevented the induction of ceramide generation by C6-ceramide, which implies ceramide generation as a possible target for the anti-apoptotic effects of Bcl-2.




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