Fig. 4: BSO and DPI cooperatively induce significant death of primary human pancreatic cancer cells.
A Comparison of cell morphology and growth in Panc-1 cells after treatment with 0.1 μM DPI, 100 μM BSO, or the combined 100 μM BSO and 0.1 μM DPI for 72 h. DPI or BSO treatment alone had a limited impact on Panc-1 cells, while the combined treatment considerably reduced the number of living cells as observed under a light microscope. Scale bar, 200 μm. B Total cellular glutathione (GSX) levels were measured by spectrophotometry after 24 h of BSO treatment at 0, 100, or 200 μM in Panc-1 cells. Treatment was performed with three independent experiments. C Western blot analysis of p22phox protein levels in Panc-1 cells and oncogenic KRASG12V-transformed HPDE cells as compared to parental HPDE cells. KDa, kilodalton. D Concurrent DPI and BSO treatment induced significant Panc-1 cell death. Cells were incubated with 0.1 or 0.3 μM DPI, 100 μM BSO, or their combination as indicated for 72 h, and cell death was determined by Annexin V/PI double staining. Panc-1 cells without any treatment served as the control. E Panc-1 cells were treated with 0.3 μM DPI and 100 μM BSO combination in the presence or absence of 2 mM NAC for 72 h. Cell death rates were determined by Annexin V/PI double staining. F Statistical analysis of the percent death rates of Panc-1 cells treated in E. Data are mean ± SD of three independent experiments with Student’s t test. ****p < 0.0001.
