Figure 5

Inhibition of Aurora-A reduces cellular resistance to cisplatin. (A) A549/DDP and H460/DDP cells were transfected with negative control (NC) or Aurora-A siRNA for 48 h and subjected to western blotting (left panel). RNAi-treated cells were incubated with indicated concentrations of DDP for 24 h, and were harvested for MTT assays (right panel). (B) A549/DDP cells expressing Aurora-A shRNA or GFP shRNA (control) under Tet-inducible system were treated cells with doxycycline (Dox, 1 μg/ml) for 48 h and harvested for western blotting analysis. Cells were subjected to colony formation assay in the presence of Dox (1 μg/ml) and DDP (0, 2, 5 μM) (n = 3), **p < 0.01, two-tailed Student’s t-tests. (C) A549/DDP cells treated with increasing doses of VX-680, MLN8237or DMSO (control) for 24 h were lysed and subjected to western blotting. (D) A549/DDP cells were treated with DDP or VX-680, or DDP in combination with VX-680 at the indicated concentrations for 24 h, and cellular viability was assessed by MTT assay (upper panel). A549/DDP cells were treated with DDP or MLN8237, or DDP in combination with MLN8237 for 24 h, and then incubated in fresh medium for another 24 h and subjected to MTT assay (lower panel). (E) A549/DDP cells with indicated treatment for 24 h were subjected to annexin V-FITC and PI staining. Data were mean ± SD (n = 3).