Effect Of G6PD On Glucose Metabolism,Growth And Migration Of Colorectal Cancer Cells And Its Correlation With HKⅡ

Objective This study was to investigate the effect of G6PD on glucose consumption,growth and migration of colorectal cancer HCT116 and SW480 cells,and the expression of G6 PD and HKⅡ in colorectal cancer and its adjacent noncancerous tissues,and their correlation.To provide an experimental basis for exploring the mechanism and early clinical diagnosis of colorectal cancer.Methods In vitro cultured colorectal cancer HCT116 and SW480 cells were subjected to G6 PD overexpression and interference treatment.Overexpression experiments were performed by Flag and Flag-G6 PD plasmids for transfection of colorectal cancer cells into Flag empty vector transfection group and Flag-G6 PD transfection group;Interference experiments were performed by transfecting cells with siRNA,and were divided into negative control group and G6PD-Homo-318,G6PD-Homo-873,and G6PD-Homo-1504 interference groups.Western blot analysis of protein expression levels of G6 PD and HKⅡ in cells;Flow cytometry analysis of cell cycle progression;Determination of glucose in culture medium by glucose oxidase method;Scratch test and Transwell test to detect cell migration and invasion ability;Immunohistochemistry was used to detect the expression levels of G6 PD and HKⅡ protein in colorectal cancer and adjacent tissues,and to analyze the correlation between them.Results 1.After overexpression of G6 PD,the expression level of G6 PD protein in Flag-G6 PD transfection group was significantly higher than that in Flag transfection group(P<0.05).After siRNA interference with G6 PD,the G6PD-Homo-318 interference group had no significant change compared with the negative control group(P>0.05),and the G6PD-Homo-873 and G6PD-Homo-1504 interference group had significantly lower G6 PD protein expression level than the negative control group.The G6PD-Homo-1504 interference is more significant.2.After overexpression of G6 PD,there was no significant change in residual glucose concentration in the two cell culture media of Flag-G6 PD transfection group compared with Flag transfection group(P>0.05).After siRNA interference with G6 PD,the residual glucose concentration in the two cell culture media of G6PD-Homo-1504 interference group was significantly higher than that of the negative control group(P<0.05).3.After overexpression of G6 PD,there was no significant change in the proportion of S cells in the Flag transfection group and the Flag-G6 PD transfection group(P>0.05).After siRNA interference with G6 PD,the proportion of S phase in the G6PD-Homo-1504 interference group was significantly lower than that in the negative control group(P<0.05).4.After overexpression of G6 PD,there was no significant difference between the two cells in the Flag transfection group and the Flag-G6 PD transfection group(P>0.05).After siRNA interference with G6 PD,the migration abilities of the two cells in the G6PD-Homo-1504 interference group were significantly lower than those in the control group(P<0.05).5.After overexpression of G6 PD,the protein expression level of HKⅡ in Flag-G6 PD transfection group was significantly higher than that in Flag transfection group(P<0.05).After siRNA interference with G6 PD,the protein expression level of HKⅡ in G6PD-Homo-1504 interference group was significantly down-regulated compared with the control group(P<0.05).6.The results of immunohistochemistry showed that the expression of G6 PD and HKⅡ in colorectal cancer tissues was significantly higher than that in adjacent tissues(P<0.05).There was a positive correlation between the expression of G6 PD and HKⅡ in colorectal cancer tissues(P<0.05).Conclusion G6PD accelerates glucose consumption in colorectal cancer cells and promotes cell proliferation and migration.Interference with G6 PD reduces the protein expression of HKⅡ,and overexpression of G6 PD promotes protein expression of HKⅡ.Compared with adjacent tissues,the expression levels of G6 PD and HKⅡ protein in colorectal cancer tissues were significantly increased,and their expression levels in colorectal cancer tissues were positively correlated.