Fig. 6: Administration of HK2 inhibitor remodels tumor vasculature to enhance blood flow, tumor perfusion, doxorubicin delivery, and efficacy against tumor growth.
a C57/BL6 mice were subcutaneously injected with 1 × 106 LLC cells and treated with placebo, 3-bromopyruvate (3-BP), doxorubicin (DOX), 3-BP, and DOX combination (n = 5 mice per group). Representative gross image of tumors from each treatment group is given. b Tumor growth curves showed that co-treatment of 3-BP and doxorubicin inhibited the growth of LLC tumors as compared with placebo, doxorubicin or 3-BP treated group. c, d Bart charts indicate the tumor weight and volume in each group. e–i Midline sections of LLC subcutaneous tumors were double immunostained for CD34 (green) and α-SMA/collagen IV (red) for blood vessel quantification, percentage of pericyte coverage, blood vessel diameter, and collagen IV intensity analysis. j LLC tumor bearing mice were intravenously injected with a FITC-PECAM antibody (green) to detect perfused blood vessels. k Bar charts show the relative area of Hoechst dye uptake into perivascular tumor cells to area of FITC-PECAM in each treatment group. l–n Ex vivo two photon microscopic imaging of FITC-PECAM antibody perfused LLC tumors after treated with DOX alone or 3-BP and DOX combination. Bar charts show the quantification of vessel diameter and doxorubicin in each group. Statistical tests were two-sided. o–p Doppler ultrasonography in live mice provides relative blood flow (peak enhancement, PE) and tumor perfusion (Wash-in perfusion index, WiPI) in subcutaneous LLC tumors. Representative ultrasound images. Bar charts, quantitation across the whole tumors, including tumor cores. Results are given as means ± SEM. b Two-way ANOVA. c, d, f–k, p One-way ANOVA. m, n Student’s t test. Scale bars in a, o represents 1 cm, e, j, k, l 50 μm, i, 20 μm.
