Figure 2.

Overexpression of SIRT6 induces ferroptosis and inhibits glycolysis in pancreatic cancer cells. (A and B) After transfection with pcDNA-SIRT6, the expression of SIRT6 in PANC-1 cells was detected by (A) reverse transcription-quantitative PCR and (B) western blotting. (C) The viability of PANC-1 cells was determined by Cell Counting Kit-8 assay. (D) Fe²⁺ content was evaluated by Fe²⁺ detection kit. (E) PANC-1 cells transfected with pcDNA-SIRT6 were treated with erastin or ferrostatin-1, and then Fe²⁺ content was evaluated by Fe²⁺ detection kit. (F) ROS level was detected by DCFH-DA method. (G) GPX4 and SLC7A11 expression levels were assessed by western blot analysis. (H-J) Glucose consumption, lactic acid and ATP production were evaluated by the corresponding kits. (K) HK2 and LDHA expression levels were detected by western blotting. ^**P<0.01 vs. control and vector group and ^#P<0.05 vs. SIRT6 group. SIRT6, sirtuin6; ROS, reactive oxygen species; GPX4, glutathione peroxidase 4; HK, hexokinase; LDHA, lactate dehydrogenase A.