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Angiopoietin-2 Overexpression in Morris Hepatoma Results in Increased Tumor Perfusion and Induction of Critical Angiogenesis-Promoting Genes

¹ Department of Anatomy and Cell Biology III, University of Heidelberg, Heidelberg, Germany; ² Clinical Cooperation Unit Nuclear Medicine, DKFZ, Heidelberg, Germany; ³ Department of Nuclear Medicine, University of Heidelberg, Heidelberg, Germany; ⁴ Department of Immunology, University of Rostock, Rostock, Germany; ⁵ Department of Radiopharmaceutical Chemistry, DKFZ, Heidelberg, Germany; and ⁶ Department of Internal Medicine III, University of Heidelberg, Heidelberg, Germany

View larger version (11K): [in a new window]   FIGURE 1.  Proliferation of HUVECs with or without (w/o) bFGF (1 ng/mL) cocultured with WT-MH3924A or Ang-2-MH3924A. Stimulation with low concentration of bFGF is not sufficient to induce proliferation of HUVECs. Note that Ang-2-MH3924A only induces proliferation in HUVEC in presence of bFGF. Values are mean ± SEM (n = 3 in duplicate; P = 0.003, Ang-2-MH3924A with vs. without bFGF; P = 0.004 vs. WT-MH3924A with bFGF).

View larger version (10K): [in a new window]   FIGURE 2.  Tumor growth in RNU rats after transplantation of WT-MH3924A or Ang-2-MH3924A until 38 d after inoculation. Values are mean ± SEM (n = 8 for each group).

View larger version (39K): [in a new window]   FIGURE 3.  H215O PET images of rats with WT- (A) and angiopoietin-2–expressing hepatoma. T = tumor.

View larger version (12K): [in a new window]   FIGURE 4.  H215O PET. Tumor tissue perfusion (K1, k2, DV, and VB values) measured in WT-MH3924A (n = 6) and Ang-2-MH3924A (n = 6) tumors of ACI rats. Ang-2–overexpressing tumors demonstrate increased K1 (A), which was not significant with P = 0.06, whereas k2 was not changed in Ang-2-MH3924A (B). Fractional volume of distribution (DV) significantly increased in Ang-2-MH3924A (C), whereas vascular fraction (VB) remained unchanged (D).

View larger version (13K): [in a new window]   FIGURE 5.  Time–activity curves for WT- and angiopoietin-2–expressing hepatomas (A). Blood pool within tumors, measured after 68Ga-DOTA-albumin application (representing permeability of vessels for albumin) showed no difference (B).

View larger version (83K): [in a new window]   FIGURE 6.  Micro- and macrovascularization of Morris hepatoma identified by CD31 or -actin staining. Representative pictures are shown. Note that Ang-2-MH3924A tumors revealed significant increase in microvessel density (as indicated by CD31 staining) compared with WT-MH3924A tumors, both in center and periphery of tumors as well as in macrovascularization (as indicated by -actin staining). (x100)