(A-D) Cells were treated with 5 μM CEH and 5 μM SP600125 alone or in combination, in addition to cDDP for 48 h; induction of apoptosis in Eca109 cells (A, B) and Eca109/CDDP (C, D) evaluated by Annexin-V-FITC/PI staining. (E-H) Cells were treated with 5 μM CEH and 0.5 μM PFTα alone or in combination, in addition to cDDP for 48 h; induction of apoptosis in Eca109 (E, F) and Eca109/CDDP (G, H) cells evaluated by Annexin-V-FITC/PI staining. (I and J) Cells were treated with 5 μM CEH, 5 μM SP600125 (I) or 0.5 μM PFTα (J) alone or in combination, in addition to cDDP for 48 h; representative images of Eca109 (upper panel) and Eca109/CDDP (lower panel) clone formation are shown. (K-N) Western blotting detection of PARP, P-gp, phosphorylated JNK (p-JNK), phosphorylated c-Jun (p-c-Jun) and Bcl-2 protein expression in Eca109 (K, M) and Eca109/CDDP (L, N) cells treated with 5 μM CEH, 5 μM SP600125 or 0.5 μM PFTα alone or in combination, in addition to cDDP for 48 h. β-Actin was used as an internal control. Data represent the mean ± SD of at least three independent experiments. Data are presented as mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001.