Sorafenib activates CD95 and promotes autophagy and cell death via Src family kinases in gastrointestinal tumor cells.
Sorafenib and vorinostat interact in a synergistic fashion to kill carcinoma cells by activating CD95; the present studies have determined how sorafenib and vorinostat individually contribute to CD95 activation. Sorafenib (3-6 micromol/L) promoted a dose-dependent increase in Src Y416, ERBB1 Y845 and CD95 Y232/Y291 phosphorylation, and Src Y527 dephosphorylation. Low levels of sorafenib-induced (3 micromol/L) CD95 tyrosine phosphorylation did not promote surface localization whereas sorafenib (6 micromol/L), or sorafenib (3 micromol/L) and vorinostat (500 nmol/L) treatment promoted higher levels of CD95 phosphorylation which correlated with DISC formation, receptor surface localization, and autophagy. CD95 (Y232F, Y291F) was not tyrosine phosphorylated and was unable to localize plasma membrane or induce autophagy. Knockdown/knockout of Src family kinases abolished sorafenib-induced CD95 tyrosine phosphorylation, DISC formation, and the induction of cell death and autophagy. Knockdown of platelet-ived growth factor receptor-beta enhanced Src Y416 and CD95 tyrosine phosphorylation, which correlated with elevated CD95 plasma membrane levels and autophagy, and with a reduced ability of sorafenib to promote CD95 membrane localization. Vorinostat increased reactive oxygen species levels, and in a delayed NF kappa B-dependent fashion, those of FAS ligand and CD95. Neutralization of FAS-L did not alter the initial rapid drug-induced activation of CD95; however, neutralization of FAS-L reduced sorafenib + vorinostat toxicity by approximately 50%. Thus, sorafenib contributes to CD95 activation by promoting receptor tyrosine phosphorylation, whereas vorinostat contributes to CD95 activation via the initial facilitation of reactive oxygen species generation and subsequently of FAS-L expression.
Park
Margaret A
MA
Department of Biochemistry Virginia Commonwealth University, Richmond, VA 23298-0035, USA.
Reinehr
Roland
R
Häussinger
Dieter
D
Voelkel-Johnson
Christina
C
Ogretmen
Besim
B
Yacoub
Adly
A
Grant
Steven
S
Dent
Paul
P
D001343:Autophagy / Q000187:drug effects*; D001557:Benzenesulfonates / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D053222:Fas Ligand Protein / Q000378:metabolism; D005770:Gastrointestinal Neoplasms / Q000201:enzymology* / Q000473:pathology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D009536:Niacinamide / Q000031:analogs & derivatives; D010671:Phenylurea Compounds; D019000:Phosphotyrosine / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D000077157:Sorafenib; D000077337:Vorinostat; D019014:fas Receptor / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
D001557:Benzenesulfonates; D053222:Fas Ligand Protein; D006877:Hydroxamic Acids; D010671:Phenylurea Compounds; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D017382:Reactive Oxygen Species; D019014:fas Receptor; D019000:Phosphotyrosine; D009536:Niacinamide; D000077337:Vorinostat; D000077157:Sorafenib; D020797:Receptor, Platelet-Derived Growth Factor beta; D019061:src-Family Kinases