A Synthetic Pan-Aurora Kinase Inhibitor, 5-Methoxy-2-(2-methoxynaphthalen-1-yl)-4H-chromen-4-one, Triggers Reactive Oxygen Species-Mediated Apoptosis in HCT116 Colon Cancer Cells

Figure 7

Effect of antioxidants on the inhibition of DK1913-induced loss of mitochondria membrane potential (ΔΨm) and activation of the caspase cascade. (a) HCT116 cells were pretreated with 100 μM butylated hydroxyanisole (BHA) for 30 min before the addition of 5 μM DK1913. After 18 h, the ΔΨm-sensitive dye JC-1 was incubated for an additional 10 min. Red and green fluorescence was visualized by an EVOSf1® fluorescence microscope (scale bar, 100 μm). (b) HCT116 cells were treated with different concentrations of BHA (0, 100, and 200 μM) for 30 min, followed by treatment with 5 μM DK1913. After 24 h, cell lysates were prepared and immunoblotting was performed using antibodies against cleaved caspase-7, caspase-3, and PARP. Arrow: proform; arrowhead: cleaved form; GAPDH: internal control. Band intensities of each blot were measured using ImageJ software and expressed relative to the basal band intensity (ser to 1) after adjusting to GAPDH levels. The data are presented as means ± SD (n = 3). NS, not significant; ; ; ; compared with the 0-time control according to Dunnett’s multiple comparisons test.

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