Figure 5 from Tumor Vessel Normalization via PFKFB3 Inhibition Alleviates Hypoxia and Increases Tumor Necrosis in Rectal Cancer upon Radiotherapy

3PO administration in combination with radiotherapy increases tumor necrosis and DNA damage in RC cells in vivo. A, Relative body weight curves of mice in different therapy groups. B, Relative growth curve of rectal PDX tumors upon treatment with vehicle (control; n = 5), 3PO (25 mg/kg; 3×/week; n = 4), RT (1.8 Gy single dose; 5×/week; n = 4), and dual therapy 3PO + RT (25 mg/kg; 3×/week; 1.8 Gy single dose; 5×/week; n = 5). Arrows indicate administration of 3PO and RT. Data are displayed as means ± SEM, ∗, P < 0.05; one-way ANOVA. ∗, P value refers to RT or RT + 3PO compared with control. C, PDX tumor weight. Data are displayed as means ± SD. ∗, P < 0.05; one-way ANOVA. D, Representative H&E staining images of PDX tumors upon treatment with vehicle (control), 3PO, RT, and 3PO + RT. Scale bars, 500 and 100 µm. Arrow heads: representative necrotic areas; asterisks: viable tumor areas. E, Viable tumor area quantification (% of total tumor area). Data are displayed as means SD. ∗, P < 0.05; ∗∗∗∗, P < 0.0001; Student t test. F, Quantification of apoptosis by cleaved caspase-3 staining. Data are displayed as means SD. ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; one-way ANOVA. G, Quantification of DNA fragmentation using TUNEL staining. Data are displayed as means SD. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗∗, P < 0.0001; one-way ANOVA. H, Representative images from TUNEL staining in PDX tumors upon treatment with vehicle (control), 3PO, RT, and 3PO + RT. Scale bars, 25 µm. I, Quantification of proliferation using bromodeoxyuridine (BrdU) staining. Data are displayed as means ± SD. ∗, P < 0.05; ∗∗∗∗, P < 0.0001; one-way ANOVA. J, Quantification of PFKFB3 expression in tumor tissue. Data are displayed as means SD. ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; one-way ANOVA. K, Representative images from PFKFB3 staining in PDX tumors upon treatment with vehicle (control), 3PO, RT, and 3PO + RT. Scale bars, 200 µm.