Fig. 5
The pharmacological treatment with BODIPY-THIF reduces angiogenesis in colorectal cancer in vitro and in vivo. A Correlations between STAT1 and angiogenesis-related markers in patients with colorectal adenocarcinoma using TCGA database. B BODIPY-THIF inhibits angiogenesis in vivo. HCT116 cells were treated with vehicle (control) or BODIPY-THIF (50 μM or 100 μM) for 48 h, and CM was collected for the CAM assay. Representative images show the appearance of blood vessels in CAM models (left panel). Images were quantitatively analyzed with WimCAM image analysis software (right panel). *p < 0.05; **p < 0.01. C STAT1 WT and STAT1 KO HCT116 cells were cultured for 48 h, and CM was collected for DIVAA. Paired angioreactors (N = 5) were recovered 14 days after implantation. Images of DIVAA angioreactors are shown (upper panel). Quantification of directed in vivo angiogenesis was assessed by FITC-dextran injection (lower panel). ***p < 0.001. D Effect of BODIPY-THIF on angiogenesis in CRC cells. HCT116 cells were treated with 5 μM Δ9-THC or 5 μM Δ9-THC plus 50 μM BODIPY-THIF for 48 h. The mRNA expression levels of the five angiogenesis-related genes were quantified by qPCR and normalized to that of GAPDH. ***p < 0.001 versus vehicle; ##p < 0.01 versus Δ9-THC; ###p < 0.001 versus Δ9-THC. E BODIPY-THIF reduces the release of VEGF from CRC cells after Δ9-THC treatment. HCT116 cells were treated with 5 μM Δ9-THC or 5 μM Δ9-THC plus 50 μM BODIPY-THIF for 48 h, and the VEGF concentration in CM was measured by ELISA. ***p < 0.001 versus vehicle; ###p < 0.001 versus Δ9-THC. F Effect of BODIPY-THIF on the binding of STAT1 to the promoters of angiogenesis-related growth factors in CRC cells. HCT116 cells were treated with 5 μM Δ9-THC or 5 μM Δ9-THC plus 50 μM BODIPY-THIF for 6 h, and the anti-STAT1 antibody was used for ChIP-qPCR to measure the binding of STAT1 to the promoters. Data were analyzed and are plotted as percent (%) of input DNA. *p < 0.05 versus vehicle; **p < 0.01 versus vehicle; #p < 0.05 versus Δ9-THC; ##p < 0.01 versus Δ9-THC