Figure 4.
In vivo angiogenesis assay characterizing antiangiogenic effects of various doses of paclitaxel. Matrigel was mixed with heparin and basic fibroblast growth factor, and was injected subcutaneously into the abdominal midline of C57BL/6 or BALB/c nude mice (a total 0.5 mg/mouse) on day 0. The mice were given 25 mg/kg of paclitaxel on days 0, 3, and 7, or 3 mg/kg or 6 mg/kg of paclitaxel every day intraperitoneally from days 0 to 10 and/or CTGF/E7 DNA vaccine on days 0 and 7. They were killed on day 11. The matrigel plugs were retrieved, resected, fixed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin to determine microvessel density (MVD). The remaining half of the matrigel plugs were assayed for hemoglobin content as described in the Materials and Methods section. (a) Representative matrigel plugs: A1, negative; A2, PBS only; A3, 3 mg/kg paclitaxel; A4, 6 mg/kg paclitaxel; and A5, 25 mg/kg paclitaxel. (b) Bar graph shows the MVDs of matrigel plugs in C57BL/6 immunocompetent mice treated with various doses of paclitaxel. The MVDs of matrigel implants in 6-mg/kg paclitaxel-treated mice were significantly lower than those in the other paclitaxel and PBS-treated mice. (c) Bar graph shows matrigel hemoglobin content for C57BL/6 immunocompetent mice treated with various doses of paclitaxel. The hemoglobin concentrations of matrigel implants in 6-mg/kg paclitaxel-treated mice were lowest among the paclitaxel-treated groups. (d) Bar graph shows matrigel hemoglobin content for BALB/c immunocompromised nude mice treated with various doses of paclitaxel: D1, negative; D2, PBS only; D3, 3 mg/kg paclitaxel only; D4, 6 mg/kg paclitaxel only; D5, 25 mg/kg paclitaxel only; and D6, 6 mg/kg paclitaxel and CTGF/E7 DNA vaccine. The hemoglobin contents of matrigel implants in the 6 mg/kg paclitaxel-treated (D4 and D6) groups were significantly lower than those in the other (D2, D3, and D5) groups in BALB/c nude mice. CTGF, connective tissue growth factor.