Fig. 4
ICG-001 inhibits OSCC growth and metastasis in vivo. a HSC-3 cells (0.5 × 106 cells/40 μl DMEM) were lentivirally transduced with DsRed and injected into tongues of 2-month-old nude mice. Tumor formation and metastasis were visualized using IVIS fluorescent imaging at day 17; representative images for mice treated with ICG-001 (n = 8; bottom) or DMSO vehicle (n = 8; top) and for control mice (n = 2) are shown. Total radiant efficiency of cells in the primary tongue tumor and of cells that metastasized to other organs was significantly reduced upon ICG-001 treatment for both the primary tongue tumor and whole body metastases (shown as the mean ± S.D.). b Caliper measurements confirmed significantly reduced primary tumor volume in ICG-001- compared to DMSO-treated mice starting at day 13 with continued trend until sacrifice at day 20. c Harvested tumors were analyzed by H&E staining for differences in morphologies. ICG-001-treated tumors exhibited a capsular phenotype, in contrast to untreated tumors that showed streaks of invasive cells growing into the tongue stroma. Fraction of the total tissue taken up by invasive epithelial cells was quantified for vehicle- and ICG-001-treated tumors (n = 3 per treatment group). d Total tissue lysates were prepared from mouse tumors (n = 4 per treatment group) and analyzed by immunoblot for E-cadherin and β-catenin protein levels after normalization to GAPDH. Immunoblot and bar graphs displaying relative protein levels are shown. e Treatment with ICG-001 alters localization of key epithelial and mesenchymal markers. Immunofluorescence imaging of E-cadherin, β-catenin, and vimentin in tumors treated with either DMSO vehicle (top) or ICG-001 (bottom). ICG-001 enhanced the recruitment of E-cadherin and β-catenin to cell-cell contacts, while reducing the expression of vimentin. *P < 0.05; **P ≤ 0.005; unpaired t test