Fig. 5: GNE-493 downregulates SphK1 protein and induces ceramide accumulation in prostate cancer cells.
From: GNE-493 inhibits prostate cancer cell growth via Akt-mTOR-dependent and -independent mechanisms
The primary prostate cancer cells, priCa-1 or priCa-2, were stimulated with GNE-493 (250 nM) or the vehicle control (0.25% DMSO, “Veh”); cells were further cultivated for an additional 12 h, the SphK1 protein expression (A), the ceramide contents (B) and the SphK1 mRNA expression (C) were tested. priCa-1 cells were infected with SphK1-expressing lentiviral particles (LV-SphK1, for 72 h) or pretreated with sphingosine 1-phosphate (S1P, 10 μM, 2 h), followed by GNE-493 (250 nM) treatment, control cells were treated with vehicle control. Cells were further cultivated for 72 h, cell viability (by testing the CCK-8 OD, E), cell death (by measuring the Trypan blue-positive cell percentage, F) and apoptosis (by examining the TUNEL-positive nuclei percentage, G) were tested. priCa-1 cells were infected with Akt1/2 shRNA lentiviral particles (Akt1/2 shRNA, for 72 h) or treated with LY294002 (1 μM), the control cells were treated with the control shRNA lentiviral particles (“shC”) plus DMSO (0.1%); Cells were cultivated for additional 12 h, levels of SphK1 protein (H) and ceramide contents (I) were tested. *P < 0.05 versus “Veh” group. #P < 0.05 versus “GNE-493” only treatment. “n.s.” stands for non-statistical difference.
