Effect of high dose vitamin C on proliferation and glucose uptake in lung cancer A549 cells
Objective To investigate the effects of high-dose vitamin C on proliferation and glucose uptake in lung cancer A549 cells.Method The IC50 value of A549 cells treated with vitamin C was analyzed by CCK8 method.Different concentrations of vitamin C(the final concentration was 0-8 mmol/L)were applied to A549 cells for 24 hours,and glucose concentrations were set at 5 mmol/L and 25 mmol/L,respectively.Cell proliferation was detected by CCK8 method.A549 cells were interfered with 8 mmol/L vitamin C,and intracellular vitamin C concentration was detected by FRASC Ascorbic acid kit Ⅱ.Human bronchial epithelioid cells(HBE cell)were used as a control to analyze the time change of intracellular vitamin C concentration for 4 h.A549 cells were then divided into four groups,and interfered with 5 mmol/L glucose,5 mmol/L glucose+8 mmol/L vitamin C,25 mmol/L glucose or 25 mmol/L glucose+8 mmol/L vitamin C.Intracellular vitamin C concentration was detected,and intracellular glucose intake was detected by using fluorescence-labeled 2-deoxyglucose as a probe.The expression levels of glucose transporters 1 and 3(GLUT1 and GLUT3)were detected by qPCR and Western Blot.Result The IC50 concentration of A549 cells treated with vitamin C for 24 h was 5.192 mmol/L.The inhibitory effect of vitamin C on A549 cell proliferation was enhanced with the increase of concentration(P＜0.05),and the cell viability of A549 cells after 5 mmol/L glucose intervention was lower than 25 mmol/L glucose when the concentration of vitamin C was 4 or 8 mmol/L(P＜0.05).Vitamin C concentration in A549 cells increased with time,and was significantly higher than that in HBE cells at 0.5,1,2 and 4 h(P＜0.05).After 8 mmol/L vitamin C intervention for 24 h,the intracellular glucose intake of A549 cells was decreased(P＜0.05),and after 25 mmol/L glucose intervention,the intracellular vitamin C concentration was lower than 5 mmol/L glucose(P＜0.05).When glucose concentration was 5 mmol/L,the expression levels of RNA and protein of GLUT1 and GLUT3 were decreased after 8 mmol/L vitamin C intervention(P＜0.05),while the RNA level of GLUT3 and the protein level of GLUT1 were decreased when glucose concentration was 25 mmol/L(P＜0.05).Conclusion At low glucose concentration,high-dose vitamin C down-regulated the expression of GLUT1 and GLUT3,decreased glucose uptake in A549 cells,and inhibited the proliferation of A549 cells.
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