Fig. 6

DKC1125 reduces the KITENIN-induced increase in aerobic glycolysis. A CaCo2/KITENIN cells were treated with the indicated concentrations of DKC1125 (0.2–5 µM) for 48 h. CaCo2/EV cells were used as controls. The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were measured using a Glycolytic Rate Assay Kit on a Seahorse XF96 extracellular flux analyzer. To assess the glycolytic rate, the assay utilizes both ECAR and OCR measurements to determine the glycolytic proton efflux rate (glycoPER). GlycoPER was measured at two time points, and then following the sequential injection of rotenone (1 μM)/antimycin A (1 μM) and 2-DG (50 mM). B Basal glycolysis and compensatory glycolysis determined using the glycoPER curves. C–D Invasion assay for CaCo/EV and CaCo2/KITENIN cells, using fibronectin as a chemoattractant. KITENIN-overexpressing cells showed greater invasiveness than empty vector-transfected cells. Cells were treated with 2.5 and 5 μM of DKC1125 for 24 h. The stained invading cells were counted and the numbers in each group are shown in a bar graph. E Mechanism whereby DKC1125 reduces the KITENIN-induced increase in glycolysis. Protein levels of c-Myc, GLUT1, HK2, PKM2, and LDHA in CaCo2/EV and CaCo2/KITENIN cells incubated for 48 h. F-G CaCo2/KITENIN cells were treated with the indicated concentrations of DKC-C14S (0.2–5 µM) for 48 h. CaCo2/EV cells were used as controls. Basal glycolysis and compensatory glycolysis determined using the glycoPER curves. H–I Cells were treated with 2.5 and 5 μM of DKC-C14S for 24 h. The stained invading cells were counted and the numbers in each group are shown in a bar graph. J Mechanism whereby DKC-C14S reduces the KITENIN-induced increase in glycolysis. Protein expression of c-Myc, GLUT1, HK2, PKM2, and LDHA in CaCo2/EV and CaCo2/KITENIN cells incubated for 48 h. Data are the mean ± standard deviation, n = 3. * p < 0.05; ** p < 0.01; *** p < 0.001, NS, no significant difference between groups