Effects Of Inhibition Of GLUT1 Expression On Glucose Metabolism And Biological Behavior In Neuroblastoma Cells

Objectives In this study,Neuroblastoma cell line SH-SY5 Y was utilized to investigate the effect of inhibiting the expression of glucose transporter 1(GLUT1)on glycolysis level and other biological behaviors such as proliferation,invasion and migration ability of tumor cells.Methods Human neuroblastoma SH-SY5 Y cells were treated with GLUT1 specific small molecule inhibitor WZB117.The expression of GLUT1 mRNA was detected by real-time fluorescent quantitative PCR.The protein expression of GLUT1,glycolysis-related enzyme and cell cyclin was detected by Western blotting.The proliferation of cells was detected by CCK-8.The content of ATP and LDH were detected by kits respectively.Cell cycle and apoptosis were detected by flow cytometry.The ability of cell invasion and migration were detected by transwell invasion and migration assay,respectively.Results 1.As the concentration of WZB117 increased,the proliferation of SH-SY5 Y cells presented a downward trend.Proliferation inhibition was most obvious at 10 ?mol/L,and the number of cell death was much more at 15 and 30 ?mol/L.Therefore,10 ?mol /L was chosen as the inhibitory concentration.2.After treating with 10 ?mol/L WZB117 for 24 h,the expression level of GLUT1 mRNA began to increase,but decreased at 72 h.From 24 h on,the expression level of GLUT1 protein was significantly decreased.3.SH-SY5 Y cells were treated with 10 ?mol/L WZB117 for 12,24,48 and 72 h respectively.The contents of ATP and LDH decreased with the extension of treatment time.4.The level of hexokinase(HK2),pyruvate kinase(PKM)and phosphofructokinase(PFKL)decreased after treating with 10 ?mol/L WZB117 for 24 h in SH-SY5 Y cells.5.After treatment of SH-SY5 Y cells with 10 ?mol/L WZB117 for 24 h,cell cycle G0-G1 phase arrest was observed,and SH-SY5 Y cells showed obvious necrosis rather than apoptosis.The expression levels of Cyclin E2,cyclin-dependent kinase(CDK2)and retinoblastoma gene pRb decreased,and the expression levels of tumor suppressor gene P53 increased.6.After treating with 10 ?mol/L WZB117 for 24 h on SH-SY5 Y cells,transwell invasion and migration experiments showed that the number of cell membrane penetration was significantly reduced.Conclusions WZB117-induced GLUT1 inhibition reduced glycolysis metabolites,suppressed tumor cell growth,induced cell cycle arrest,and inhibited cell invasion and migration in NB cells in vitro.This study suggested that GLUT1 can be used as a potential therapeutic target for NB.