Phospholipase D protects ECV304 cells against TNFalpha-induced apoptosis.
Tumor necrosis factor alpha (TNFalpha), a pleiotropic cytokine, activates both apoptotic and pro-survival signals depending on the cell model. Using ECV304 cells, which can be made TNFalpha-sensitive by cycloheximide (CHX) co-treatment, we evaluated the potential roles of ceramide and phospholipase D (PLD) in TNFalpha-induced apoptosis. TNFalpha/CHX induced a robust increase in ceramide levels after 16 h of treatment when cell death was maximal. PLD activity was increased at early time point (1h) whereas both PLD activity and PLD1 protein were strongly decreased after 24h. TNFalpha/CHX-induced cell death was significantly lowered by exogenous bacterial PLD and phoshatidic acid, and in cells overexpressing PLD1. Conversely, cells depleted in PLD proteins by small interference RNA (siRNA) treatment exhibited higher susceptibility to apoptosis. These results show that PLD exerts a protective role against TNFalpha-induced cell death.
Birbes
Helene
H
INSERM U585 and INSA-Lyon and IMBL, Bldg Louis Pasteur,11 avenue Jean Capelle, F-69621 Villeurbanne Cedex, France.
Zeiller
Caroline
C
Komati
Hiba
H
Némoz
Georges
G
Lagarde
Michel
M
Prigent
Annie-France
AF
D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D002518:Ceramides / Q000235:genetics; D003513:Cycloheximide / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D010739:Phospholipase D / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D020011:Protective Agents; D034741:RNA, Small Interfering / Q000494:pharmacology; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D002518:Ceramides; D020011:Protective Agents; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D003513:Cycloheximide; D010739:Phospholipase D; C106801:phospholipase D1