Targeting Non-Small Cell Lung Cancer Cells by Dual Inhibition of the Insulin Receptor and the Insulin-Like Growth Factor-1 Receptor (original) (raw)
Figure 3
Inhibition of the IGF1R and IR in combination does not inhibit growth of A549 cells more effectively than inhibition of IGF1R alone.
(A) Comparison of expression of IGF1Rβ and IRβ in Hcc193 and A549 cell lines was determined by western blotting using 15 µg protein extracted from each cell line. (B) A549 cells were cultured under anchorage–independent conditions in 0.1% serum in the presence of 3GF alone and in combination with αIR3 (2 µg/ml), IR47-9 (2 µg/ml), αIR3 (2 µg/ml) and IR47-9 (2 µg/ml), and AZ12253801 (50 nM), as indicated. After 5 days viable cells were estimated by the fluorescence of the metabolic reduction product of Alamar blue. Each bar represents the mean fluorescence of four replicate wells ± standard deviation from a single experiment. The results shown are representative of 3 independent experiments. ***, p<1×10−6; **, p<0.0001; *, p<0.05 (valid for all repeats); two-tailed unpaired t test. (C) A549 cells were treated with αIR3 (2 µg/ml), IR47-9 (2 µg/ml), αIR3 (2 µg/ml) and IR47-9 (2 µg/ml), and AZ12253801 (50 nM) as indicated for 2 h before 10 min incubation with vehicle or 3GF. Phosphorylation of Akt on S473 and expression of IGFRβ and IRβ was determined by western blotting. An anti-α-tubulin antibody was used as a control for protein loading. Scanned images of immunoblots are cropped to feature the prominent band.