Figure 6
Cells expressing KRASG12V were sensitive for glycolytic inhibition in vitro and in vivo. (A,B) CaCO2-control, CaCO2-BRAFV600E, CaCO2-KRASG12V, HT29 and SW480 cells were grown in physiological (1.0 g/L) glucose concentrations and treated with 0.25 mM 3-Bromopyruvate (BrPy), 1 mM L-glyceraldehyde (L-GA) or PBS (−) for 24 h. Viable cells compared to PBS are depicted. Shown are standard deviation of (in minimum) n = 3 replicates. p < 0.05 was indicated with asterisk (unpaired two-tailed t Test). (C) Phosphorylation of signaling molecules were analyzed with ELISA bead-based phosphoproteomics technology (Luminex). Shown are log2 fold changes (fc) to PBS. (D) CaCO2-BRAFV600E and CaCO2-KRASG12V treated with 0.25 mM BrPy (+) or PBS (−) for 24 h were analyzed with antibodies against phospho-AMPK (Thr172). Vinculin served as loading control. Representative images are shown. Full-length blots are shown in the Supplementary Information. (E) CaCO2-KRASG12V cells were subcutaneously injected into the right flank of mice receiving 200 µL PBS or BrPy (8 mg/kg) in the presence of Doxycycline treatment every 2 days (starting from day 8) by intraperitoneal injection. Mean of tumor volume is shown for indicated mice per group up to 15 d. Shown are standard deviation of n = 5 (control group) and n = 10 (BrPy group) replicates. p < 0.05 was indicated with asterisk (unpaired two-tailed t Test).
