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. 2017 Jan 14;8(8):12941–12952. doi: 10.18632/oncotarget.14652

Figure 4. Effects on 93T449 and SW872 cells of combined treatment with CDK4 and IGF1R inhibitors.

Figure 4

(A and B) IGF1R amplification and expression in 93T449 and SW872 cells, as well as an IGF1R-amplified tumor (DDLPS_25T). Relative copy number was estimated by NGS and quantitative PCR (qPCR) (A). mRNA expression was estimated by quantitative RT-PCR (qRT-PCR) and normalized to GAPDH expression (B). Relative expression levels are expressed as ratios of the median expression in non-amplified tumor samples, as in Figure 2B. (C and D) Growth inhibitory effects of CDK4 and IGF1R inhibitors on 93T449 (C) and SW872 (D) cells. Palbociclib (CDK4 inhibitor) and NVP-AEW541 (IGF1R inhibitor) were added at various concentrations, and cell metabolic activities were assayed after 6 days of culture. In this assay, synergism of these inhibitors was evaluated using CompuSyn (http://www.combosyn.com) [41]. Their effects were synergistic in 93T449 cells (average combination index score = 0.42 ± 0.19), but not in SW872 cells (average combination index score = 6.28 ± 3.22). (E) Effect of IGF1R inhibitor on IGF1R phosphorylation in 93T449 and SW872 cells. Cells were treated with NVP-AEW541 (1 μM) for 12 or 24 h and harvested. Expression and Y1135 phosphorylation of IGF1R were evaluated by western blotting analysis.