Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

J. Cell Sci. 115 (23): 4671-4683

Research Article

Caspase-dependent initiation of apoptosis and necrosis by the Fas receptor in lymphoid cells: onset of necrosis is associated with delayed ceramide increase

Claudio A. Hetz1, Martin Hunn2, Patricio Rojas1, Vicente Torres1, Lisette Leyton1, and Andrew F. G. Quest1,*

1 Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile
2 Institute of Biochemistry, University of Lausanne, Switzerland

* Author for correspondence (e-mail:aquest{at}

Accepted for publication 4 September 2002.

Abstract: Engagement of the Fas receptor promotes apoptosis by activation of caspases. In addition, alterations in plasma membrane lipid orientation and intracellular ceramide levels are often observed. In A20 B-lymphoma cells, FasL-induced cell death and phosphatidylserine (PS) externalization were completely prevented by the generic caspase inhibitor z-VAD-fmk. By contrast, the caspase-3 inhibitor Ac-DEVD-cho only partially restored cell viability and had no effect on surface exposure of PS. Flow cytometric analysis after FasL treatment identified two populations of dead cells. In one, death was dependent on caspase-3 and paralleled by DNA fragmentation and cell shrinkage. In the second, death occurred in the absence of caspase-3 activity and apoptotic features but was also blocked by zVAD-fmk. By morphological criteria these were identified as apoptotic and necrotic cells, respectively. Using fluorescent substrates, caspase-3 activity was detected only in the apoptotic cell population, whereas caspase-8 activity was detected in both. Both forms of caspase-8-dependent cell death were also detected downstream of Fas in Jurkat T-cells, where Fas-dependent PS externalization and delayed ceramide production, which is similar to results shown here in A20 cells, have been reported. However, for Raji B-cells, lacking lipid scrambling and ceramide production in response to Fas activation, only apoptosis was detected. Short-chain C2- or C6-ceramides, but not the respective inactive dihydro compounds or treatment with bacterial sphingomyelinase, induced predominantly necrotic rather than apoptotic cell death in A20 B-, Raji B- and Jurkat T-cells. Thus, delayed elevation of ceramide is proposed to promote necrosis in those Fas-stimulated cells where caspase-8 activation was insufficient to trigger caspase-3-dependent apoptosis.

Key Words: Lymphoid cells • Fas • Apoptosis • Necrosis • Caspases • Ceramide

Ceramide and mitochondrial function in aging oocytes: joggling a new hypothesis and old players.
L. L. Kujjo and G. I. Perez (2012)
Reproduction 143, 1-10
   Abstract »    Full Text »    PDF »
Age-Dependent Increases in Apoptosis/Necrosis Ratios in Human Lymphocytes Exposed to Oxidative Stress.
M. I. Behrens, M. Silva, A. Schmied, F. Salech, H. Manzur, R. Rebolledo, R. Bull, V. Torres, M. Henriquez, and A. F. G. Quest (2011)
J Gerontol A Biol Sci Med Sci 66A, 732-740
   Abstract »    Full Text »    PDF »
Helicobacter pylori--Induced Loss of the Inhibitor-of-Apoptosis Protein Survivin Is Linked to Gastritis and Death of Human Gastric Cells.
M. Valenzuela, G. Perez-Perez, A. H. Corvalan, G. Carrasco, H. Urra, D. Bravo, H. Toledo, and A. F. G. Quest (2010)
The Journal of Infectious Disease 202, 1021-1030
   Abstract »    Full Text »    PDF »
Biological Roles of Acid and Neutral Sphingomyelinases and Their Regulation by Nitric Oxide.
C. Perrotta and E. Clementi (2010)
Physiology 25, 64-71
   Abstract »    Full Text »    PDF »
A. J. Zullo, M. Michaud, W. Zhang, and M. J. Grusby (2009)
J. Biol. Chem. 284, 12504-12511
   Abstract »    Full Text »    PDF »
Bioactive sphingolipids: metabolism and function.
N. Bartke and Y. A. Hannun (2009)
J. Lipid Res. 50, S91-S96
   Abstract »    Full Text »    PDF »
Changes in mitochondrial dynamics during ceramide-induced cardiomyocyte early apoptosis.
V. Parra, V. Eisner, M. Chiong, A. Criollo, F. Moraga, A. Garcia, S. Hartel, E. Jaimovich, A. Zorzano, C. Hidalgo, et al. (2008)
Cardiovasc Res 77, 387-397
   Abstract »    Full Text »    PDF »
E-Cadherin Is Required for Caveolin-1-Mediated Down-Regulation of the Inhibitor of Apoptosis Protein Survivin via Reduced {beta}-Catenin-Tcf/Lef-Dependent Transcription.
V. A. Torres, J. C. Tapia, D. A. Rodriguez, A. Lladser, C. Arredondo, L. Leyton, and A. F. G. Quest (2007)
Mol. Cell. Biol. 27, 7703-7717
   Abstract »    Full Text »    PDF »
Casein kinase 2 (CK2) increases survivin expression via enhanced beta-catenin-T cell factor/lymphoid enhancer binding factor-dependent transcription.
J. C. Tapia, V. A. Torres, D. A. Rodriguez, L. Leyton, and A. F. G. Quest (2006)
PNAS 103, 15079-15084
   Abstract »    Full Text »    PDF »
Caveolin-1 controls cell proliferation and cell death by suppressing expression of the inhibitor of apoptosis protein survivin.
V. A. Torres, J. C. Tapia, D. A. Rodriguez, M. Parraga, P. Lisboa, M. Montoya, L. Leyton, and A. F. G. Quest (2006)
J. Cell Sci. 119, 1812-1823
   Abstract »    Full Text »    PDF »
Role of membrane sphingomyelin and ceramide in platform formation for Fas-mediated apoptosis.
M. Miyaji, Z.-X. Jin, S. Yamaoka, R. Amakawa, S. Fukuhara, S. B. Sato, T. Kobayashi, N. Domae, T. Mimori, E. T. Bloom, et al. (2005)
J. Exp. Med. 202, 249-259
   Abstract »    Full Text »    PDF »
Increased peripheral platelet destruction and caspase-3-independent programmed cell death of bone marrow megakaryocytes in myelodysplastic patients.
E. J. Houwerzijl, N. R. Blom, J. J. L. van der Want, H. Louwes, M. T. Esselink, J. W. Smit, E. Vellenga, and J. Th. M. de Wolf (2005)
Blood 105, 3472-3479
   Abstract »    Full Text »    PDF »
Endoplasmic reticulum stress-induced programmed cell death in soybean cells.
A. Zuppini, L. Navazio, and P. Mariani (2004)
J. Cell Sci. 117, 2591-2598
   Abstract »    Full Text »    PDF »
Caspase-12 and endoplasmic reticulum stress mediate neurotoxicity of pathological prion protein.
C. Hetz, M. Russelakis-Carneiro, K. Maundrell, J. Castilla, and C. Soto (2003)
EMBO J. 22, 5435-5445
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882