Vorinostat and sorafenib increase ER stress, autophagy and apoptosis via ceramide-dependent CD95 and PERK activation.
We recently noted that low doses of sorafenib and vorinostat interact in a synergistic fashion to kill carcinoma cells by activating CD95, and this drug combination is entering phase I trials. The present studies mechanistically extended our initial observations. Low doses of sorafenib and vorinostat, but not the individual agents, caused an acidic sphingomyelinase and fumonisin B1-dependent increase in CD95 surface levels and CD95 association with caspase 8. Knock down of CD95 or FADD expression reduced sorafenib/vorinostat lethality. Signaling by CD95 caused PERK activation that was responsible for both promoting caspase 8 association with CD95 and for increased eIF2alpha phosphorylation; suppression of eIF2alpha function abolished drug combination lethality. Cell killing was paralleled by PERK-and eIF2alpha-dependent lowering of c-FLIP-s protein levels and overexpression of c-FLIP-s maintained cell viability. In a CD95-, FADD- and PERK-dependent fashion, sorafenib and vorinostat increased expression of ATG5 that was responsible for enhanced autophagy. Expression of PDGFRbeta and FLT3 were essential for high dose single agent sorafenib treatment to promote autophagy. Suppression of PERK function reduced sorafenib and vorinostat lethality whereas suppression of ATG5 levels elevated sorafenib and vorinostat lethality. Overexpression of c-FLIP-s blocked apoptosis and enhanced drug-induced autophagy. Thus sorafenib and vorinostat promote ceramide-dependent CD95 activation followed by induction of multiple downstream survival regulatory signals: ceramide-CD95-PERK-FADD-pro-caspase 8 (death); ceramide-CD95-PERK-eIF2alpha- downward arrowc-FLIP-s (death); ceramide-CD95-PERK-ATG5-autophagy (survival).
Park
Margaret A
MA
Department of Biochemistry, Virginia Commonwealth University, Richmond, VA 23298-0035, USA.
Zhang
Guo
G
Martin
Aditi Pandya
AP
Hamed
Hossein
H
Mitchell
Clint
C
Hylemon
Philip B
PB
Graf
Martin
M
Rahmani
Mohamed
M
Ryan
Kevin
K
Liu
Xiang
X
Spiegel
Sarah
S
Norris
James
J
Fisher
Paul B
PB
Grant
Steven
S
Dent
Paul
P
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001343:Autophagy; D001557:Benzenesulfonates / Q000494:pharmacology*; D053181:Caspase 8 / Q000378:metabolism; D002470:Cell Survival; D002518:Ceramides / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008954:Models, Biological; D009154:Mutation; D009536:Niacinamide / Q000031:analogs & derivatives; D010671:Phenylurea Compounds; D011725:Pyridines / Q000494:pharmacology*; D000077157:Sorafenib; D016896:Treatment Outcome; D000077337:Vorinostat; D019892:eIF-2 Kinase / Q000096:biosynthesis*; D019014:fas Receptor / Q000096:biosynthesis*
D000970:Antineoplastic Agents; D001557:Benzenesulfonates; D053181:Caspase 8; D002518:Ceramides; D015852:Eukaryotic Initiation Factor-2; D006877:Hydroxamic Acids; D009536:Niacinamide; D010671:Phenylurea Compounds; D011725:Pyridines; D000077157:Sorafenib; D000077337:Vorinostat; D019892:eIF-2 Kinase; D019014:fas Receptor
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