Study On Detection Of Tumor Cell Metabolites And Antitumor Properties Of β-glucan

Glioblastoma(GBM)cells have abnormally elevated levels of glycolysis,suggesting that the glycolytic pathway may be a potential therapeutic target.The PI3K-mTOR signaling pathway,locating downstream of the epidermal growth factor receptor(EGFR),not only participates in the regulation of cell survival and proliferation,but also plays an important role in the metabolism of substances and energy in cells.At present,there is no single cell level research for the glycolysis of GBM cells and the antitumor mechanism of EGFR inhibitors.In addition,the anti-tumor research of polysaccharides has been a hot spot for anti-tumor research of natural active substances.However,there are few studies on β-glucan antitumor in vivo.In this paper,an animal model will be established to analyze the anti-tumor effects and possible mechanisms of β-glucan in vivo.In this study,single cell barcode chip(SCBC)technology was used to detect the glucose uptake in GBM cells at the single cell level.The Hill equation can be used to fit the relationship between fluorescence intensity and glucose-biotin(Gluc-Bio);By using a redox reaction,which Resazurin(non-fluorescent)was restored to Resorufin(red fluorescent)by lactic acid,lactate dehydrogenase and diaphorase catalyze,the quantitative detection of lactic acid was achieved.Hill equation can also be used to fit the relationship between fluorescence intensity of Resorufin and the amount of lactic acid metabolism.In addition,after erlotinib treatment,glucose uptake and lactic acid production in GBM39 cells were significantly reduced(P<0.001);at the same time,the phosphorylation levels of p70S6 K,EGFR,mTOR,ERK,NDRG1,Src,4EBP1,Ki67,and Akt were also significantly decreased(P<0.001),indicating that erlotinib inhibit the glycolysis process of GBM cells,and then the phosphorylation levels of Src-Ras-Raf-MEK-ERK and PI3K-mTOR pathway related proteins downstream of EGFR were decreased.In this paper,we also used S180 ascites tumor mice as an animal model to analyze the anti-tumor activity and possible mechanism of yeast β-glucan in vivo.The results showed that the tumor volume and quality of mice with S180 ascitic fluid tumor significantly decreased,and the tumor inhibition rate significantly increased after treatment with yeast β-glucan.The further analysis of anti-tumor mechanism revealed that S180 ascites tumor mice spleen index,serum CD4 and CD8 levels increased,CD4/CD8 ratio decreased,while serum TNF-α,IL-2 and IL-6 levels also increased after treatment with yeast β-glucan.FITC-PI double staining combined with flow cytometry and Western blotting analysis showed that yeast β-glucan treatment can induce apoptosis of tumor cells in mice with S180 ascitic fluid tumor,and the ratio of Bax/Bcl-2 in apoptosis cells was increased.The above results indicate that the anti-tumor mechanism of yeast β-glucan may be related to the enhancement of host immune function and induction of tumor cell apoptosis in vivo.