Fig. 3. Combination of gefitinib and APAP induced accelerated ROS generation, resulting in a DNA damage response. (A-E) HL-7702 cells were treated with 10 μM gefitinib and/or 1.25 mM APAP for 24 h. (A) A ROS assay kit showed ROS generation in HL-7702 cells. (B) The expression levels of γH2AX and ACTB were determined by western blot. (C) Typical images of DNA damage after treatment detected by comet assay. Scale bar=50 μm. (D) Quantitative analysis was performed on the percentage of DNA tail, n=50 cells per group. (E) HL-7702 cells treated with 10 μM gefitinib and 1.25 mM APAP were combined with or without 1 mM NAC for 24 h. The expression levels of c-PARP and ACTB were determined by western blot. All experiments were performed in triplicate. For western blots, one of three similar experiments is shown, densitometric analysis was carried out. Data are presented as the mean ± SEM (n=3); n.s=no significance; **p<0.01; ***p<0.001. APAP, acetaminophen; GEFI, gefitinib; NAC, N-acetyl-L-cysteine.
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