Effects of interfering Twist gene on glycolysis in pancreatic cancer PANC1 cells and its mechanism / 中华胰腺病杂志

ABSTRACT

Objective To observe the effects of interfering Twist expression on glycolysis in pancreatic cancer PANC1 cells and explore the potential mechanism.Methods Twist targeting siRNA (siTWIST) was designed and synthesized,and pancreatic cancer PANC1 cells were transfected with siTWIST using liposome,with nonspecific siRNA (siNC) transfected cells as negative control group and untransfected cells as blank control group.RT-PCR and Western blot were used to detect the expression levels of Twist mRNA and protein,and Akt and p-Akt protein expression in different groups;cell proliferation was detected by MTT;and glycolysis key enzyme (pyruvate kinase and hexokinase) activity and the content of lactic acid in the supernatant of culture fluid were detected.Results Twist mRNA level in control group,siRNA group and siTwist group expression was 1.00 ± 0.09,1.01 ± 0.08 and 0.36 ± 0.02;Twist protein level was 0.41 ± 0.05,0.42 ± 0.04 and 0.12 ± 0.06;the mRNA and protein expression in siTWIST group was obviously lower than those in control group and the difference was statistically significant (P ＜ 0.05).The cell survival rate was (100.02± 9.36)％,(100.01 ± 10.25)％ and (59.32± 7.26)％,which in siTWIST group was obviously lower than that in control group;pyruvate kinase activity was (0.73 ± 0.05)U/mg,(0.72 ± 0.08) U/mg and (0.43 ± 0.05) U/mg;the activities of hexokinase were (4.98 ± 0.48) U/mg,(4.96 ± 0.52) U/mg and (2.54 ± 0.21) U/mg;the content of lactic acid was (20.36 ± 2.61) mmol/L,(20.64 ± 3.05) mmol/L and (9.48 ± 1.09) mmol/L;the ratio of p-Akt/Akt was 0.65 ± 0.04,0.63 ± 0.07,and 0.25 ± 0.04,which in siTWIST group was obviously lower than that in control group.Conclusions Interference with Twist expression could inhibit the proliferation and glycolysis pathway in pancreatic cancer cells,and the mechanism may be related to the Akt signaling pathway.