SUPPLEMENTAL RESULTS Long-term treatment of HCT116.hCG.Luc orthotopic tumors: in vivo monitoring and survival To confirm our previous results in a second colon cancer cell line, thirty-one mice

Long-term treatment of HCT116.hCG.Luc orthotopic tumors: in vivo monitoring and survival To confirm our previous results in a second colon cancer cell line, thirty-one mice were intracecally implanted with HCT116.hCG.Luc. Two weeks later, mice were randomized according to bioluminescence signal and treatments with (1) control vehicle, (2) FOLFOX-like, (3) topotecan, (4) topotecan+pazopanib, (5) FOLFOX-like+topotecan, (6) FOLFOX-like+pazopanib, (7) FOLFOX-like+topotecan+pazopanib (n=4-5) were initiated. As pazopanib monotherapy did not show any effect in our previous results, this group was not included in this experiment. Furthermore, UFT was replaced by a combination of UFT+Folinic acid+oxaliplatin, in order to closely mimic the clinical standard therapy of FOLFOX. This experiment lasted 28 weeks (supplemental Figures 4 and 5). At this endpoint, eight mice (2 mice on topotecan, 4 mice on topotecan+pazopanib, and 2 mice on FOLFOX-like+topotecan+pazopanib) were still alive with very limited tumor burden. FOLFOX-like, as well as the combination of FOLFOX-like+pazopanib did not significantly delay the increase in bioluminescence signal compared to controls (supplemental Figure 4), but did reduce the absolute strength of bioluminescence signal (supplemental Figure 5A). In contrast, all groups treated with a regimen containing oral topotecan showed a delayed increase and reduced maximum level in bioluminescence. Animals treated with topotecan alone, FOLFOX-like+topotecan, topotecan+pazopanib and FOLFOX-like+topotecan+pazopanib showed a significantly reduced signal compared to control animals (P<0.05). Among these groups, no significant difference in bioluminescence was observed. These results were confirmed by quantification of secreted β-hCG (supplemental Figure 5B). β-hCG-levels were lower in animals treated with