Fig. 6.

Effects of NS-398 on subcutaneous SK-N-AS tumor development. Three days after sc inoculation of SK-N-AS cells (2.5×10⁶ cells), mice received daily injections of NS-398 (15 mg/kg, ip) or vehicle (control) for the following 5 weeks (n=5 for each group). (A): Development of sc SK-N-AS tumors. Data are expressed as mean±SE (n=5). *Significantly different from control (p<0.05). (B) Tumor wet weight at 5 weeks after sc inoculation. Treatment with NS-398 significantly inhibited tumor wet weight. Data are expressed as mean±SE (n=5). *Significantly different from control (p<0.05). (C) Immunohistochemical examination of subcutaneous SK-N-AS tumors. Tumor sections were stained with anti-factor VIII antibody and anti-VEGF-A antibody as described in Materials and Methods. Tumors from NS-398-treated mice showed a decrease in microvessel density (factor VIII staining) and reduced intensity of VEGF-A staining compared to control. Negative control: no primary antibody. Original magnification, x50 (factor VIII); x200 (VEGF-A). (D) Effects of NS-398 on VEGF-A mRNA expression in SK-N-AS tumor in vivo. One µg of total RNA from sc tumors was subjected to RT-PCR analysis in duplicate. NS-398 down-regulated hVEGF-A₁₂₁ mRNA.