Figure 6
PI3K/HSP90/CHIP/ILK/ERK1/2/NF-κB signaling determines cell migration and controls susceptibility to the anticancer agents 5-FU and cisplatin. (A) Migration activity was tested in shLuc- or shCHIP-transfected AGS cells treated with LY294002 (25 μM) and 17-AAG (0.2 μM) for 12 h. shLuc, control siRNA, or DMSO were used as negative controls. The numbers of migrated cells in the observed field are shown. PI staining followed by flow cytometry analysis showed 5-FU- or cisplatin-induced apoptosis for 2 days in shILK- or siIQGAP1-transfected AGS cells (B), AGS cells pretreated with the indicated inhibitors for 0.5 h (C), and shLuc- or shCHIP-transfected AGS cells treated with LY294002 or 17-AAG (D). shLuc, control siRNA, or DMSO were used as negative controls. Plasmids based on pcDNA3.1-Flag-ILK containing full-length ILK (ILK1-452), fragment 171-452 (ILK171-452), or fragment 1-170 (ILK1-170) were transfected into MKN45 cells. Compared with pcDNA3.1-Flag (Flag) only, cell migration (E) and apoptosis induced by 5-FU (5 μM) (F) were detected in the absence or presence of PD98059 and CAPE. Data are mean ± SD from three independent experiments. *P <0.05, **P <0.01 and ***P <0.001 compared with control. #P <0.05, ##P <0.01, and ###P <0.001 compared with control. NS: not significant.