Figure 5

Chemosensitive ovarian cancer cells are more sensitive to glucose deprivation. (A) A2780 and (B) C200 cells were cultured in glucose free (0mM glucose) Roswell Park Memorial Institute 1640 media containing 5% dialyzed fetal bovine serum or supplemented with 10 mM glucose or 1 mM pyruvate. Proliferation of cells was measured using CYQUANT® NF cell Proliferation Assay by measuring fluorescence intensity at different time points as indicated. (C) Bar graph representation of the cell proliferation under various conditions at day 3. The experiment was performed in triplicates and repeated twice. ***p < 0.001, **p < 0.01 C200 compared to A2780. Under similar conditions, at 48 h, an equal number of cells were plated in multi-well Seahorse Biosciences XF96 plate (Boston, MA) for measurement of ECAR (D,E) and OCR (H,I). For ECAR, injections 1, 2, 3 represents glucose, oligomycin and 2DG respectively. For OCR, injections 1,2,3 represent oligomycin, FCCP and rotenone respectively. Basal ECAR (F), glycolytic capacity (G), basal OCR (J) and maximum respiration (K) were calculated as described in the Materials and Methods and are represented as bar graphs. The experiment was performed in triplicates and repeated twice. ***p < 0.001, **p < 0.01 C200 compared to A2780. (I) Under similar conditions ATP production was measured by a Biovision fluorescence assay kit. ***p < 0.001, **p < 0.01, C200 compared to A2780.