Antibiotics · January 27, 2022

(B) Flow cytometric analysis of untreated SKOV3 or SKOV3/DDP cells

(B) Flow cytometric analysis of untreated SKOV3 or SKOV3/DDP cells. of highly elevated ROS. Furthermore, the inhibition of Bcl-2 reduced the OXPHOS and sensitivity of SKOV3/DDP cells to cisplatin in a selective manner. Furthermore, when combined with 2-deoxyglucose (2-DG), the anticancer effect of the Bcl-2 inhibitor ABT737 was greatly potentiated and hypoxia-inducible factor 1 (HIF-1) appeared to be closely associated with Bcl-2 family members in the regulation of glucose metabolism. These results suggested that the special glucose metabolism in SKOV3/DDP cells might be selectively targeted by disrupting Bcl-2-dependent OXPHOS. (5). As expected, SKOV3/DDP cells exhibited considerable resistance to cisplatin, while SKOV3 cells also exhibited resistance to cisplatin as determined by the MTT assay following exposure to increasing concentrations of cisplatin for 24 h (Fig. 1A). As shown in Fig. 1B, SKOV3/DDP cells were preferentially enriched for G0/G1 quiescent cells and had a lower proliferation rate. The expression of genes associated with glucose metabolism was assessed by RT2 Human Glucose Metabolism Profiler PCR array. The obtained results indicated the upregulation of glycolysis, the tricarboxylic acid cycle (TCA) cycle and gluconeogenesis in D-Luciferin sodium salt SKOV3/DDP cells (Fig. 1C and Table II). Open in a separate window Figure 1 Glucose metabolism is altered in cisplatin-resistant cells. (A) The cells were EPHB2 subjected to various doses of cisplatin for 24 h prior to being evaluated by MTT assay. Data are presented as the mean standard deviation, n=3. (B) Flow cytometric analysis of untreated SKOV3 or SKOV3/DDP cells. The percentage of cells in the G0/G1, S, or G2/M phases of the cell cycle was indicated. (C) The expression of glucose metabolism-related genes (84 genes) was evaluated in D-Luciferin sodium salt cells using a human glucose metabolism polymerase chain reaction array. The changes in gene expression are indicated in the heat map. Red indicates upregulation (SKOV3/DDP vs. SKOV3), and green indicates downregulation. The names and positions of the genes name are listed in the table. DDP, cisplatin. Table II Functional grouping of gene expression. and as well as elevated glycogen levels (Fig. 2D). As glycogen is a branched polymer of glucose that acts as an intracellular glucose store, high glycogen levels may render the cells less sensitive to glucose deprivation (Fig. 2E). Notably, SKOV3/DDP cells exhibited reduced sensitivity to glucose deprivation compared with SKOV3 cells (Fig. 2F), while the combined treatment with 2-DG (glycolysis inhibitor) induced significant cell death compared with the glucose deprivation alone group (Fig. 2G). Open in a separate window Figure 2 Cisplatin-resistant cells exhibit a higher demand for glucose. (A) The D-Luciferin sodium salt glucose uptake of SKOV3 or SKOV3/DDP cells was determined using the glucose analogue 2-NBDG. **P 0.01 vs. SKOV3 cells. (B) Glucose consumption and (C) lactate production were measured in the culture media using glucose and lactate kit and normalized to the protein content. *P 0.05, **P 0.01 vs. SKOV3 cells. (D) Expression levels of glycolytic genes were determined using quantitative polymerase chain reaction. The genes were normalized to -actin. **P 0.01 vs. SKOV3 cells. (E) Glycogen levels were determined using a glycogen kit. **P 0.01 vs. SKOV3 cells. (F) The effects of glucose deprivation on cell viability were determined by MTT assay. The data are presented as the percentage of cell number compared with the control group and as the mean standard deviation (n=3). **P 0.01 vs. control. (G) The effects of glucose deprivation combine with 10 mM 2-DG on cell viability in two cell lines. **P 0.01 vs. SKOV3 cells. ##P 0.01 vs. glucose deprivation group. DDP, cisplatin; PFKL, liver phosphofructokinase; PDK1, pyruvate dehydrogenase kinase 1; LDHA, lactate dehydrogenase.