Antioncogenic Effects of Transient Receptor Potential Vanilloid 1 in the Progression of Transitional Urothelial Cancer of Human Bladder
Figure 3
Capsaicin stimulates invasiveness of 5637 UCCs. Total and active MMP9 levels were evaluated in UCC lysates after capsaicin treatment by MMP9 activity assay. Statistical analysis was performed by comparing capsaicin with untreated cells, normalized for the relative vehicle, * (a). MMP9 levels were evaluated in UCCs treated with capsaicin alone or in combination with different doses of pAB. Statistical analysis was performed by comparing capsaicin-treated UCCs with capsaicin plus pAB-treated UCCs, * (b). Cell invasion was evaluated in UCCs treated with capsaicin in combination with pAB, by Matrigel invasion assay. Statistical analysis was performed by comparing capsaicin plus pAB-treated UCCs with capsaicin-treated UCCs, * (c). GZMA expression was evaluated in untreated and capasicin-treated UCCs by using a FITC-conjugated anti-human GZMA mAb and cytofluorimetric analysis. Numbers in the corner represent the mean fluorescence intensity. Gray areas represent the negative control (d). GZMA release was evaluated in the supernatant of untreated and capsaicin-treated UCCs by ELISA. Statistical analysis was performed by comparing capsaicin-treated with untreated UCCs, normalized with the relative vehicle treatments, * (e). Microtubule disassembly was evaluated by confocal microscopy using anti-human α-tubulin and FITC-conjugated goat Abs in vehicle or capsaicin-treated UCCs. Colchicine treatment was performed as positive control. Bar = l0 μM (f). Total protein lysates from UCCs treated with capsaicin or vehicle were separated by electrophoresis and probed with anti-human α-tubulin followed by horseradish peroxidase-conjugated goat anti-mouse Abs (g).
