Fig. 5. Knockdown of FGF2 in NPC tumor cells enhances AAD-mediated anti-tumor effects.

A–D Tumor growth (A, B) and tumor weights (C) were measured in shScrambled- and shFGF2-transfected NPC tumors receiving vehicle or anti-VEGF treatment. The tumor inhibition ratios were calculated (D) (n = 6 mice per group). E Representative micrographs of Ki67⁺ proliferative cells and cleaved caspase-3⁺ apoptotic cells in vehicle- or anti-VEGF-treated shScrambled- and shFGF2-transfected NPC tumors. Scale bar = 50 μm. Quantification of Ki67⁺, cleaved caspase-3⁺ signals, and PA index in vehicle- or anti-VEGF-treated shScrambled- and shFGF2-transfected NPC tumors (n = 8 random fields per group). F Representative micrographs of CD31⁺ microvessels and CA9⁺ hypoxic areas in vehicle- or anti-VEGF-treated shScrambled- and shFGF2-transfected NPC tumors. Scale bar in upper panel = 100 μm, scale bar in lower panel = 50 μm. Quantification of CD31⁺ tumor vessel parameters and CA9⁺ signals in vehicle- or anti-VEGF-treated shScrambled- and shFGF2-transfected NPC tumors (n = 8 random fields per group). G Blood samples from tumor-bearing mice were FACS analyzed for EGFP⁺ signals. Quantification of EGFP⁺ circulating tumor cells (n = 6 samples per group). H Quantification of clones after culturing blood samples from vehicle- or anti-VEGF-treated shScrambled- and shFGF2-transfected NPC-bearing mice for two weeks (n = 6 samples per group). I Representative graphs of lungs from vehicle- or anti-VEGF-treated shScrambled- and shFGF2-transfected NPC-bearing mice. Arrows indicate visible metastatic nodules. Scale bar = 0.5 cm. EGFP⁺ metastatic signals in the lung. Quantification of pulmonary metastasis proportion (n = 6 mice per group). **P < 0.01; ***P < 0.001. NS not significant. Data presented as mean ± SD.