Fig. 5

High expression of BID associates with sensitivity to SAC abrogation in cancer cell lines of diverse origins. A Dose-response plots to AZD2811 (upper panels) and BAY1217389 (lower panels) of cancer cell lines of lung, prostate, breast, pancreas, ovarian, colon and other origins. Values shown are means ± SD of ≥2 independent experiments. In each experiment, every concentration of drug was tested in six wells (n = 6). B BID mRNA levels in cancer cell lines of diverse origins. Cells classified according to response to SAC abrogation in S, sensitive (n=5) and R, resistant (n=14) to both AURKBi and TTKi. Each point represents a cell line, lines indicate mean ± SD. ****p<0.0001 (Student’s t test). C Western blotting analysis of BID, MDM2 and CASP-3 protein levels in cancer cell lines of diverse origins. The seven cell lines sensitive to AURKBi and/or TTKi, together with eight resistant cell lines, were selected for the analysis. Cells sensitive and resistant to both AURKBi and TTKi are indicated in green and blue, respectively. The two cell lines sensitive exclusively to one of the inhibitors are indicated in grey. D BID, MDM2 and CASP3 protein levels in cancer cell lines of diverse origins, quantified from the Western blotting image presented in (C). The intensity of the bands was normalized using β-tubulin. Cells are classified according to response to SAC abrogation in S, sensitive (n=5) and R, resistant (n=14) to both AURKBi and TTKi. Each point represents a cell line, lines indicate mean ± SD. **p<0.001; ns=not significant (Student’s t test). E Cell cycle analysis of the NCI-H1819, MDA-MB-468, SK-MES-1 and WM793 cultures. Cells were allowed to attach, serum starved for 24 h, FBS (10%) and AZD2811 (150 nM) or BAY1217389 (50 nM) were added and cultures submitted to PI staining at the indicated times. Lines sensitive (S) and resistant (R) to SAC abrogation are indicated in green and blue, respectively. F Percentage of apoptotic/necrotic cells in NCI-H1819, MDA-MB-468, SK-MES-1 and WM793 cultures. Cells were treated with AZD2811 (150 nM) or BAY1217389 (50 nM) and cultures submitted to annexin V/PI staining at the indicated times. Bars indicate mean ± SD of two independent experiments. *p<0.05 (Student’s t test). Lines sensitive and resistant to SAC abrogation are indicated in green and blue, respectively