Inhibition of nitric-oxide-mediated apoptosis in Jurkat leukemia cells despite cytochrome c release.
We have recently shown that nitric-oxide (NO)-induced apoptosis in Jurkat human leukemia cells requires degradation of mitochondria phospholipid cardiolipin, cytochrome c release, and activation of caspase-9 and caspase-3. Moreover, an inhibitor of lipid peroxidation, Trolox, suppressed apoptosis in Jurkat cells induced by NO donor glycerol trinitrate. Here we demonstrate that this antiapoptotic effect of Trolox occurred despite massive release of the mitochondrial protein cytochrome c into the cytosol and mitochondrial damage. Incubation with Trolox caused a profound reduction of intracellular ATP concentration in Jurkat cells treated by NO. Trolox prevented cardiolipin degradation and caused its accumulation in Jurkat cells. Furthermore, Trolox markedly downregulated the NO-mediated activation of caspase-9 and caspase-3. Caspase-9 is known to be activated by released cytochrome c and together with caspase-3 is considered the most proximal to mitochondria. Our results suggest that the targets of the antiapoptotic effect of Trolox are located downstream of the mitochondria and that caspase activation and subsequent apoptosis could be blocked even in the presence of cytochrome c released from the mitochondria.
Umansky
V
V
Division of Cellular Immunology, German Cancer Research Center, D-69120, Heidelberg, Germany. V.Umansky@dfkz-heidelberg.de
Ratter
F
F
Lampel
S
S
Bucur
M
M
Schirrmacher
V
V
Ushmorov
A
A
D000255:Adenosine Triphosphate / Q000378:metabolism; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000502:physiology*; D002308:Cardiolipins / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D002469:Cell Separation; D002839:Chromans / Q000494:pharmacology*; D003574:Cytochrome c Group / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D014810:Vitamin E / Q000031:analogs & derivatives
D000975:Antioxidants; D002308:Cardiolipins; D002839:Chromans; D003574:Cytochrome c Group; D014810:Vitamin E; D009569:Nitric Oxide; D000255:Adenosine Triphosphate; D020169:Caspases; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid