Figure 4.

IGF1R inhibition synergizes with isiPI3K in inhibiting the viability of HNC^HPV+ cells. (A) UM-SCC47 and UT-SCC60A sensitive and acquired resistance cells (Res2) were cultured with DMSO, GDC0032 (500 nM) or BYL719 (2 μM), AEW541 (1 μM), or a combination of AEW541 with GDC0032 or BYL719. Four days post-treatment, cell viability was determined using crystal violet staining. The proliferation experiment was repeated 3 times, and representative results from one experiment are presented. Biological replicates from one experiment are shown as grey dots. (B) ZIP model synergy test was performed in both UM-SCC47 and UT-SCC60A cell lines. Cells were cultured with increasing concentrations of GDC0032 (0 up to 2 μM) or BYL719 (0 up to 10 μM) and AEW541 (0 up to 10 μM). Four days post-treatment cell viability was determined using crystal violet stanning. The results are presented as a ZIP synergy score with 3-D surface plots displaying synergy regions (left panel) along with a dose-response matrix (right panel). The synergy experiment was repeated 3 times, and representative results from one experiment are presented. (C) UM-SCC104, UT-SCC102, and UPCI-SCC90 sensitive cells were cultured with DMSO, BYL719 (2 μM), AEW541 (1 μM), or a combination of the two drugs. Four days post-treatment cell viability was determined using crystal violet staining. The proliferation experiment was repeated 3 times, and the results of one representative experiment are presented as means ± SEM. Biological replicates from one experiment are shown as grey dots. Statistical significance was calculated by one-way ANOVA, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.