Figure 3

Quantitative analyses of metastasis, vascular density, apoptosis, cell proliferation in mammary carcinomas. A. Multiplicity of lymph node metastasis was significantly decreased in the pesVEGFR-2 and pEndo groups (**P < 0.01 as compared to the pVec group; †P < 0.01 as compared to the pesVEGFR-2 group). B. Multiplicity of lung metastatic foci > 250 μm was significantly reduced in the pesVEGFR-2 and pEndo groups (**P < 0.01 as compared to the pVec group; †P < 0.01 as compared to the pesVEGFR-2 group). C. Quantitation of blood and lymphatic microvessels were conducted using CD34 and Lyve-1 immunohistochemistry. The number of CD34⁺/Lyve-1^- blood vessels was significantly decreased in the pEndo group, but not in the pesVEGFR-2 group, as compared to the pVec group. The number of CD34^-/Lyve-1⁺ lymphatic microvessels was significantly decreased in the pesVEGFR-2 and pEndo groups. D. The mammary tumors were immunohistochemically stained for another lymphatic endothelial marker podoplanin. The number of dilated lymphatic microvessels containing intraluminal tumor cells was significantly lower in pesVEGFR-2 and pEndo groups than those observed in the control pVec group. E. Apoptotic cell death, assessed by TUNEL assay, was significantly increased in the pEndo group. F. Cell proliferation, inferred by BrdU labeling indices, was significantly decreased in the pesVEGFR-2 and pEndo groups. *P < 0.05 and **P < 0.01 as compared to the values of the pVec group. Data presented are means ± SD values.