The Effect And Machanism Of Cytoplasmic Macrophage Cololy-stimulating Factor(M-CSF) On Glucose Metabolism In Human Breast Cancer MCF-7 Cells

Objective:To explore the effect and mechanisms of cytoplasmic M-CSF on glucose metabolism in human breast cancer MCF-7 cells.Methods:MCF-7 cells stable expressing cytoplasmic M-CSF were constructed in the laboratory. Relative ATP content was measured by ATP assay kit. Glucose was measured using Glucose assay kit,and Lactate was measured using Lactate Acid assay kit.The expression of HK2,PKM2 and GLUT-1 was detected by Western bloting,The effect of LY294002 or API-2 on expression of GLUT-1. The sensitivity of MCF-7 and MCF7-M cells to 5-FU before and after ATP depletion by 3-Br PA pretreatment was observed by MTT assay.Results:MCF-7-M cells displayed significantly higher intracellular ATP levels compared with MCF-7 cells(p<0.05),treatment of MCF-7 cells with the hexokinase inhibitor 2-DG led to a drop in ATP(p<0.05), as well as treatment of MCF-7-M cells with 2-DG led to a prominent drop in ATP(p<0.01),whereas the mitochondrial ATPase inhibitor oligomycin A had limitedeffects(p>0.05). MCF-7-M cells consumed glucose or produced lactate at a higher rate than MCF-7 cells. Glucose and lactate is decreased with API-2 treatment in MCF-7 cells and MCF-7-M cells(p<0.01).The results from western blot showed that the expression level of HK2,PKM2,GLUT-1 are higher in MCF-7-M cells compared with control cells and LY294002 or API-2 significantly inhibits the expression of GLUT-1 in MCF-7-M cells(p<0.05).From MTT assays show pretreatment of 3-Br PA sensitized MCF-7 and MCF-7-M cells to 5-FU(p<0.01).Conclusion:Cytoplsmic M-CSF activates aerobic glycolysis by induce GLUT-1 、 Hexokinase2 and Pyruvate Kinase M2 protein expression in MCF-7 cells, PI3K/Akt signaling involves that glycolysis was activated by Cytoplsmic M-CSF in MCF-7 cells.