The Mechanism Of IL-37-mediated Glycolysis Of Lung Adenocarcinoma By Inhibiting GLUT1 Expression

Objective: Adenocarcinoma of lung is one of the most common causes of human death.The treatment of advanced lung adenocarcinoma is limited and the therapeutic effect is not satisfactory.Recent studies have shown that glycolysis plays an important role in the development of lung adenocarcinoma.Previous studies have found that Interleukin 37(IL-37)can inhibit glycolysis in A549 cells.However,the mechanism of IL-37 affecting lung adenocarcinoma glycolysis has not been clarified.The purpose of this study was to investigate whether IL-37 affects glycolysis of lung adenocarcinoma A549 cells by inhibiting the expression of glucose transporter 1(GLUT1)and to elucidate the mechanism.Methods: Quantitative real-time Polymerase Chain Reaction(qRT-PCR)and Western blot were used to detect the expression of IL-37 in lung adenocarcinoma A549 cells(A549 cells)and normal lung epithelial BEAS-2B cells(BEAS-2B cells).q RT-PCR was used to detect the m RNA expression of GLUT1,Pyruvate kinase M(PKM)in A549 cells and BEAS-2B cells;Western blot was used to detect the protein expression of GLUT1 and PKM2 in A549 cells and BEAS-2B cells.A549 cells were treated with recombinant Human IL-37(rh IL-37)at concentration 100ng/m L,q RT-PCR and Western blot were used to detect the expression of GLUT1 and PKM2 in A549 cells.Recombinant lentivirus was used to overexpress IL-37 in A549 cells,q RT-PCR and Western-blot were used to detect GLUT1 expression before and after IL-37 overexpression in A549 cells.A549 cells were treated with BAY-876 at different concentrations(0μM,2 μM,4 μM,6 μM,8 μM,10 μM),Western blot was used to detect the expression of GLUT1 in A549 cells.A549 cells were treated with BAY-876 at 6 μM concentration,further glucose and lactic acid content were detected in A549 cell culture fluid.Research data was analyzed using the statistical software in Graph Pad Prism 9.Differences with P < 0.05 were considered statistically significant.Results: qRT-PCR and Western blot results showed that IL-37 expression was significantly lower in A549 cells than BEAS-2B cells(P<0.01).q RT-PCR results showed that GLUT1 and PKM m RNA expression was significantly higher in A549 cells than BEAS-2B cells(P<0.01).Western blot results showed that GLUT1 protein expression were higher in A549 cells than BEAS-2B cells(P<0.05),PKM2 protein expression were significantly higher(P<0.01).q RT-PCR results showed that the GLUT1 m RNA expression in A549 cells was lower than A549 cells treated with rh IL-37(P<0.05).Western blot results showed that the GLUT1 protein expression in A549 cells was lower than A549 cells treated with rh IL-37(P<0.05).There was no significant difference in PKM2 m RNA expression in A549 cells before and after rh IL-37 treatment.q RT-PCR and Western blot results showed that GLUT1 m RNA and protein expression had no significant difference before and after IL-37 had overexpressed in A549 cells.A549 cells were treated with BAY-876 at different concentrations(0μM,2μM,4μM,6μM,8μM and10μM)for 24 h.Western blot results showed that GLUT1 expression was significantly decreased in A549 cells with 6μM concentration BAY-876(P<0.01).Compared with A549 cells without BAY-876 treatment,glucose content in cell culture fluid of A549 cells treated with 6u M BAY-876 was significantly increased,while lactic acid content was significantly decreased(P<0.01),which indicates that glucose uptake and lactic acid production significantly decreased.Conclusion: Endogenous overexpression of IL-37 could not inhibit the expression of GLUT1 in A549 cells.Exogenous administration of IL-37 can inhibit glucose uptake and lactic acid production by inhibiting GLUT1 expression,thus affecting glycolysis of lung adenocarcinoma A549 cells and playing a role in glucose metabolism of lung adenocarcinoma cells.This study provides new ideas for the study of lung adenocarcinoma and glucose metabolism,and provides new targets for the treatment of lung adenocarcinoma.