ZD55-AChE induced gastric cancer cell apoptosis through mitochondrial pathway. (A) Observation of the nucleic fragmentation in adenovirus-infected AGS cells by Hoechst333258 staining assay. Arrows indicated fragmented nucleic. Scale bar: 200 μm. (B) Measurement of the cell cycle distributions of adenovirus-infected AGS cells by PI staining. Numbers are the proportion of cells in sub-G1 phase. (C) Alteration of the proportion of Annexin V positive cells by adenoviruses. Numbers in the first and fourth quadrants are the proportion of apoptotic cells. (D) ZD55-AChE changed mitochondrial membrane potential (Δψm) of AGS cells. Numbers in the trapezoid region indicate the ratio of cells with disrupted Δψm. (E) Western blot of apoptosis-associated proteins. GAPDH was used as an internal control. (F) Caspase inhibitor Z-VAD-FMK alleviated the cytotoxicity of ZD55-AChE. The results are expressed as mean ± SD (n = 3). ***P < 0.001, NS: non-significant. AGS cells were infected with the indicated adenovirus at a MOI of 2 for 48 hours (A-F) or 72 hours (E).