Cross-talk between ceramide and PKC activity in the control of apoptosis in WEHI-231.
WEHI-231, a murine B-cell lymphoma, readily undergoes programmed cell death following surface immunoglobulin (Ig) cross-linking [1]. Ceramide has been shown to induce apoptosis in WEHI-231 following its exposure to anti-lg antibodies, dexamethasone, and irradiation [2]. Recently, Haimovitz-Friedman et al. have demonstrated in endothelial cells that PMA not only prevented ceramide mediated apoptosis, but inhibited the generation of ceramide following irradiation [3]. In this paper we use highly specific PKC inhibitors to explore the connection between PKC activity, ceramide signaling and apoptosis. Both chelerythrine chloride and calphostin C triggered rapid apoptosis in WEHI-231 and acted in synergy with exogenous ceramide to induce apoptosis. Detailed studies of chelerythrine's mechanism of action revealed that 30 minutes following addition of 10 microM chelerythrine, sphingomyelin and phosphatidylcholine (PC) mass decreased confirming our previous findings of neutral, but not acidic, sphingomyelinase activation following treatment with PKC inhibitors [4]. The novel observation that inhibition of PKC isoforms present in WEHI-231 leads to a rapid rise in cellular ceramide as a results of sphingomyelin hydrolysis further suggests an antagonistic relationship between PKC activity and ceramide in the signaling events preceding apoptosis.
Department of Pathology, University of Chicago, Illinois, USA.
S J
SJ
Chmura
E
E
Nodzenski
M A
MA
Crane
S
S
Virudachalam
D E
DE
Hallahan
R R
RR
Weichselbaum
J
J
Quintans
D000470:Alkaloids; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053119:Benzophenanthridines; D004357:Drug Synergism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016393:Lymphoma, B-Cell / Q000473:pathology; D051379:Mice; D009281:Naphthalenes / Q000494:pharmacology; D010617:Phenanthridines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D015398:Signal Transduction / Q000502:physiology*; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000470:Alkaloids; D053119:Benzophenanthridines; D004791:Enzyme Inhibitors; C064769:N-acetylsphingosine; D009281:Naphthalenes; D010617:Phenanthridines; D013109:Sphingomyelins; C061465:calphostin complex; C016299:chelerythrine; D011493:Protein Kinase C; D013108:Sphingomyelin Phosphodiesterase; D013110:Sphingosine