Mechanism of epigallocatechin gallate combined with cholecystokinin receptor inhibitor to inhibit the growth of pancreatic cancer cells
HU Li-juan,ZHANG Ai-min,GE Li-xiu
Abstract:
Objective To investigate the inhibitory effect and molecular mechanisms of epigallocatechin gallate (EGCG) on pancreatic cancer cells alone or in combination with cholecystokinin receptor (CCKBR) inhibitors. Methods The control group, EGCG group, YF476 group, and the combination group of EGCG and YF476 (E+Y group) were respectively treated with the control medium, 50 μmol/L EGCG, 0.1 nmol/L YF476, and 50 μmol/L EGCG and 0.1 nmol/L YF476 for 24 h and 48 h on pancreatic cancer MiaPaCa-2 cells. The proliferation ability of cells in each group was detected by CCK-8 method. The changes in glycolysis of cells in each group were detected by glucose-lactate kit. The expressions of glycolytic enzymes and proteins in the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway and CCKBR pathway in each group of cells were detected by Western blot method. Results The results of CCK-8 showed that EGCG or YF476 alone could inhibit the proliferation of MiaPaCa-2 cells, and the combined effect of the two was more significant than that alone (P <0.05). Glucose consumption and lactate production experiments showed that either EGCG or YF476 alone could inhibit glycolysis in MiaPaCa-2 cells, and the combined effect of the two was more significant than that alone (P <0.05). The results of Western blot showed that EGCG could inhibit the expression of glycolytic enzyme and phosphorylated PI3K and AKT, and the inhibitory effect of EGCG combined with YF476 was more significant than that alone (P <0.05), and the expression of glycolytic enzyme, CCKBR and phosphorylated PLC was inhibited by YF476 alone, and the inhibitory effect of EGCG combined with YF476 on the expression of glycolytic enzyme, CCKBR and their pathway proteins was more obvious (P <0.05). YF476 alone could inhibit the expression of glycolytic enzymes, CCKBR and phosphorylated PLC, and the inhibitory effect of EGCG combined with YF476 was more significant than that alone (P <0.05). Conclusion Both EGCG and cholecystokinin receptor inhibitor can inhibit the proliferation and glycolysis of human pancreatic cancer cells MiaPaCa-2, the combined effect of them is more significant than that of alone, and the mechanism may be related to the inhibition of PI3K/AKT and CCKBR pathway coupling.
