Induction of Lysosomal Membrane Permeabilization Is a Major Event of FTY720-Mediated Non-Apoptotic Cell Death in Human Glioma Cells

SIMPLE SUMMARY: FTY720, a sphingosine-1-phosphate (S1P) analog, is a potent immunosuppressant for the treatment of multiple sclerosis. In addition to being an immune modulator, FTY720 also features antitumor activity in several cancer models, but the molecular mechanisms are unclear. Here, we extended our research to analyze the signaling pathways mediating FTY720-induced cell death. FTY720 did not induce apoptotic cell death, autophagy, paraptosis, or necroptosis in glioma cells. Interestingly, FTY720 accumulated in lysosomes, resulting in the induction of lysosomal membrane permeabilization (LMP). Inhibition of LMP by overexpression of HSP70 and cathepsin inhibitors blocked FTY720-induced cell death. These data suggest that FTY720 induces cell death induced by LMP in glioma cells. ABSTRACT: FTY720, a sphingosine-1-phosphate (S1P) receptor modulator, is a synthetic compound produced by the modification of a metabolite from I. sinclairii. Here, we found that FTY720 induced non-apoptotic cell death in human glioma cells (U251MG, U87MG, and U118MG). FTY720 (10 µM) dramatically induced cytoplasmic vacuolation in glioma cells. However, FTY720-mediated vacuolation and cell death are not associated with autophagy. Genetic or pharmacological inhibition of autophagy did not inhibit FTY720-induced cell death. Herein, we detected that FTY720-induced cytoplasmic vacuoles were stained with lysotracker red, and FTY720 induced lysosomal membrane permeabilization (LMP). Interestingly, cathepsin inhibitors (E64D and pepstatin A) and ectopic expression of heat shock protein 70 (HSP70), which is an endogenous inhibitor of LMP, markedly inhibited FTY720-induced cell death. Our results demonstrated that FTY720 induced non-apoptotic cell death via the induction of LMP in human glioma cells.