Fig. 4: GNE-493 induces oxidative stress and programmed necrosis 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 designated time, ROS contents (by examining CellROX intensity, A), lipid peroxidation (by examining TBAR intensity, B), DNA break (by examining ssDNA contents, C) and mitochondrial depolarization (tested by green JC-1 monomers accumulation, D) were tested. Mitochondrial immunoprecipitation (“Mito-IP”) assays were employed to examine the association of p53-CyPD-ANT1 (in priCa-1 cells, E); expression of p53-CyPD-ANT in mitochondrial lysates was tested by Western blotting assays (E, “Inputs”). Cell necrosis was tested by measuring medium LDH release (F). The primary human prostate cancer cells, priCa-1 or priCa-2, were pretreated for 30 min with N-acetylcysteine (NAC, 400 μM) or the CyPD inhibitor cyclosporin A (CsA, 10 μM), or stably infected with CyPD shRNA lentivirus (sh-CyPD), followed by GNE-493 (250 nM) treatment and cultivated for 72 h, cell viability and cell death were then separately examined by CCK-8 (G) and the Trypan blue staining (H) assays. *P < 0.05 versus “Veh” group. #P < 0.05 versus “GNE-493” only treatment. Scale bar = 100 μm (A and D).
