Fig. 9
PFKFB4-knockdown overcomes Sunitinib resistance in ccRCC in vivo. A) Xenograft murine models consisting of 8 male BALB/c nude mice per group with subcutaneous implanted Sun-sensitive A498 cells with or without PFKFB4-KD (shRNA#2) under right hind limb with tumor growth monitored over 60-day period and tumor size of < 2500 mm3 as endpoint, with representative tumor image at endpoint (bar = 1 cm) analyzed by two-way ANOVA and B) Kaplan-Meier curves of survival of mice, analyzed by Log-rank test; C) Representative immunohistochemical staining of factors in tumors from A), with intensity scored semi-quantitatively and statistically compared; D) Xenograft murine models consisting of 8 male BALB/c nude mice per group with subcutaneous implanted Sun-resistant 786O cells fed with 20 mg/kg of Sun and/or 120 mg/kg of 5MPN orally by gavage; tumor growth monitored over 60-day period, analyzed by two-way ANOVA and E) Kaplan-Meier curves of survival of mice, analyzed by Log-rank test; F) Representative immunohistochemical staining CD31 targeting micro-vessels in tumors (red arrows) from D), with relative micro-vessel density (MVD) statistically compared; G) Tail vein injection of Sun-resistant A498 cells in 9 mice per group with 5MPN, Sun or combo treatments (Tx); mice monitored for 4 weeks for photon detection, normalized to PreTx CON group; bar figures showing photon change before (PreTx) and after (PostTx) with representative luciferase image showing lung involvement at endpoint of each group H) Schematic cartoon of regulatory axis proposed by the current study. (All in vitro assays performed in triplicates and at least 3 biological replicates; ns = not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)