Table 3. Micrometastatic B16LS9 VEGF and PEDF mRNA levels after supplemental angiostatin

In vivo the level of VEGF and PEDF mRNA (mean±SEM) in hepatic micrometastases was determined by real time RT-PCR. When B16LS9 cells were grown in vitro in the presence of the supplemental angiostatin prior to implantation, the level VEGF and PEDF mRNA and ratio of VEGF to PEDF mRNA level (mean±standard deviation) in hepatic micrometastases was less than when they were grown in media alone. VEGF and PEDF mRNA values are normalized to β-actin according to Pfaffl's mathematical model for relative quantification in real time PCR [23].

  Groups        VEGF/PEDF     VEGF/β-actin    PEDF/β-actin
-----------   -------------   -------------   -------------
Angiostatin   0.0274±0.0070   0.0593±0.0172   2.1492±0.1292
PBS           0.1726±0.0313   0.0516±0.0135   0.3000±0.0573
-----------   -------------   -------------   -------------
p value       0.0014          0.5782          0.00002