Re-evaluation of the distinction between type I and type II cells: the necessary role of the mitochondria in both the extrinsic and intrinsic signaling pathways upon Fas receptor activation.
Cyclosporin A (CyA) and bongkrekic acid (BK) prevented Fas-induced apoptosis in two type I cell lines (H9 and SKW6.4) and two type II cell lines (Jurkat and CEM). CyA and BK inhibited the release of cytochrome c in all four cell lines. In type I cells and in CEM cells, CyA and BK did not prevent the translocation of Bax to the mitochondria. In these same cells, full-length Bid decreased in the mitochondria and cytosol. The cleavage product of Bid, tBid, appeared in the cytosol and to a lesser extent in the mitochondria. In Jurkat cells, Bid also decreased in the cytosol, but increased in the mitochondria. Similar to the other cells, tBid appeared in the mitochondria and cytosol. In the type I H9 and SKW6.4 cells and type II Jurkat cells, the caspase-8 inhibitor Z-Ile-Glu(OMe)-Thr-Asp(OMe)-CH2F (IETD) prevented the cell killing. In the type I cells, IETD prevented the translocation of Bax, the degradation of Bid and the accumulation of tBid. By contrast, IETD only marginally protected the type II CEM cells. In these cells in the presence of IETD, Bax translocated to the mitochondria, in the absence of any degradation of Bid or accumulation of tBid. In the type I H9 cells, IETD produced a depletion of ATP, an effect that did not occur in the type II CEM cells. It is concluded that in type I cells the extrinsic signaling pathway is mitochondrial dependent to the same extent as is the intrinsic pathway in type II cells.
Tafani
Marco
M
Department of Experimental Medicine and Pathology, La Sapienza University, Rome, Italy.
Karpinich
Natalie O
NO
Serroni
Ada
A
Russo
Matteo A
MA
Farber
John L
JL
D000911:Antibodies, Monoclonal; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001865:Bongkrekic Acid / Q000494:pharmacology; D053181:Caspase 8; D061945:Caspase Inhibitors; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival; D016572:Cyclosporine / Q000494:pharmacology; D045304:Cytochromes c / Q000037:antagonists & inhibitors; D003600:Cytosol / Q000187:drug effects / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology; D015398:Signal Transduction; D019014:fas Receptor / Q000187:drug effects / Q000502:physiology*
D000911:Antibodies, Monoclonal; D061945:Caspase Inhibitors; D009842:Oligopeptides; D019014:fas Receptor; C425644:isoleucyl-glutamyl-threonyl-aspartic acid fluoromethyl ketone; D001865:Bongkrekic Acid; D016572:Cyclosporine; D045304:Cytochromes c; C505530:CASP8 protein, human; D053181:Caspase 8