Figure 4 from Tumor Vessel Normalization via PFKFB3 Inhibition Alleviates Hypoxia and Increases Tumor Necrosis in Rectal Cancer upon Radiotherapy

3PO potentializes irradiation effects in colorectal cancer cells. A, Colony formation assay performed with HCT-116, HT-29, SW-1463, and SW-837 cancer cells upon 3PO treatment. Cells treated with DMSO or 2.5 µmol/L 3PO for 4 hours and exposed to indicated doses of RT. Six days (HCT-116), 7 days (HT-29), and 14 days (SW-1463 and SW-837) later, colonies were stained and counted. Data are displayed as means ± SD, n = 3. ∗, P < 0.05; one-way ANOVA. B, Colony formation assays performed with HCT-116, HT-29, SW-1463, and SW-837 cells treated with different concentrations of 3PO for 4 hours, exposed to either RT (6 Gy) or not irradiated and analyzed after 6 days (HCT-116), 7 days (HT-29), 14 days (SW-1463), and 14 days (SW-837) by counting colonies. C, Invasion assay performed with HCT-116, HT-29, SW-1463, and SW-837 cancer cells. Cells were plated in a Boyden multi-well chamber and treated with 10 µmol/L 3PO for 4 hours, subjected to RT (6Gy), and incubated for a period of 96 hours. Data are displayed as means SD, n = 3. ∗∗, P < 0.01; ∗∗∗∗, P < 0.0001; one-way ANOVA. D–F Cell migration assay quantification of HCT-116, HT-29, and SW-837 cancer cells upon combinatory treatment. Cells were treated with 3PO for 4 hours and subjected to RT (6 Gy). Data are displayed as means ± SD, n = 3. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; one-way ANOVA.