Fig 2.
Structural features observed by electron microscope (10000× and 5000×) in PC3, DU145, and LNCaP prostate cancer cells treated with KML001 for 24 and 48 h.
Fig 3.
Induction of apoptosis by KML001 in prostate cancer cells.
(A) FACS analysis of annexin V/PI staining. Results show early apoptosis, defined as annexin V-positive and PI-negative cells, and late apoptosis, defined as annexin V-positive and PI-positive cells. Results were expressed as means ± SD of three independent experiments. (B) Western blot analysis of the time- and dose-dependent cleavage of PARP and activation of procaspase-3.
Fig 4.
Induction of autophagy by KML001 in prostate cancer cells.
(A) Western blot analysis of the time- and dose-dependent conversion of LC3-I to-II. (B) Inhibition by 3-MA of KML001-induced conversion of LC3 in prostate cancer cells. (C) Cells were exposed to 10 μM (PC3), 5 μM (DU145), or 2 μM (LNCaP) KML001 in the presence or absence of 1 mM 3-MA for 72 h. Results were expressed as means ± SD of three independent experiments. * p < 0.05 by one-way ANOVA.
Fig 5.
Regulation of autophagy and apoptosis by ROS.
All 3 prostate cancer cells were treated with the indicated concentration of KML001 in the absence or presence of 5 mM NAC for 24 h. (A) KML001 induces dose-dependent ROS (blue) accumulation. Cells were stained with DCFH-DA and washed with PBS. More than three fields in each cell were observed by fluorescence microscope (200×), and representative images are shown. (B) NAC inhibition of KML001-induced conversion of LC and caspase activation in prostate cancer cells. (C) Cells were exposed to 10 μM (PC3), 5 μM (DU145), or 2 μM (LNCaP) KML001 in the presence or absence of 1 mM NAC for 72 h. Results were expressed as means ± SD of three independent experiments. * p < 0.05 by one-way ANOVA.