Park Margaret A, Zhang Guo, Martin Aditi Pandya, Hamed Hossein, Mitchell Clint, Hylemon Philip B, Graf Martin, Rahmani Mohamed, Ryan Kevin, Liu Xiang, Spiegel Sarah, Norris James, Fisher Paul B, Grant Steven, Dent Paul
Department of Biochemistry, Virginia Commonwealth University, Richmond, VA 23298-0035, USA.
Cancer Biol Ther. 2008 Oct;7(10):1648-62. doi: 10.4161/cbt.7.10.6623. Epub 2008 Oct 12.
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).
我们最近注意到,低剂量的索拉非尼和伏立诺他以协同方式相互作用,通过激活CD95来杀死癌细胞,并且这种药物组合已进入I期试验。目前的研究从机制上扩展了我们最初的观察结果。低剂量的索拉非尼和伏立诺他,而非单独使用其中一种药物,会导致酸性鞘磷脂酶和伏马菌素B1依赖性的CD95表面水平升高以及CD95与半胱天冬酶8的结合增加。敲低CD95或FADD的表达会降低索拉非尼/伏立诺他的致死性。CD95发出的信号导致PERK激活,这既促进了半胱天冬酶8与CD95的结合,也导致eIF2α磷酸化增加;抑制eIF2α功能可消除药物组合的致死性。细胞杀伤伴随着PERK和eIF2α依赖性的c-FLIP-s蛋白水平降低,而c-FLIP-s的过表达维持了细胞活力。索拉非尼和伏立诺他以CD95、FADD和PERK依赖性方式增加ATG5的表达,这导致自噬增强。PDGFRβ和FLT3的表达对于高剂量单药索拉非尼治疗促进自噬至关重要。抑制PERK功能会降低索拉非尼和伏立诺他的致死性,而抑制ATG5水平则会提高索拉非尼和伏立诺他的致死性。c-FLIP-s的过表达阻断了细胞凋亡并增强了药物诱导的自噬。因此,索拉非尼和伏立诺他促进神经酰胺依赖性的CD95激活,随后诱导多个下游生存调节信号:神经酰胺-CD95-PERK-FADD-前半胱天冬酶8(死亡);神经酰胺-CD95-PERK-eIF2α-↓c-FLIP-s(死亡);神经酰胺-CD95-PERK-ATG5-自噬(存活)。