The Effect Of Tumor-associated Macrophage On The Proliferation,Migration And Glycolysis Of Gastric Cancer Cells

Objective: Gastric cancer is the most common malignant tumor in the digestive tract with a high fatality rate.In recent years,more and more studies have found that the tumor microenvironment is closely related to the occurrence and development of tumors.As an important part of the tumor microenvironment,tumor-associated macrophage(TAM)can promote the proliferation of tumor cells,and promote tumor cell migration and invasion by inducing epithelial-mesenchymal transition.However,the specific mechanism of TAM interacting with gastric cancer cells to regulate the growth of cancer cells is still unclear.Therefore,in this study,we mainly explore the effects of TAM on the biological characteristics of gastric cancer cells and the possible regulatory mechanism,which may provide new ideas and research directions for improving clinical treatment.Methods:1.THP-1 cells were induced by Phorbol myristate acetate(PMA)and IL-4/IL-13.We detected the expression of CD206,FN1 and CCL22 by Real-time qPCR.Cellular immunofluorescence was used to detect the expression of CD163 and CD206 on the TAM surface.2.TAM supernatant culture medium was collected to make conditioned medium to treat HGC-27,NCI-N87 cells,and the proliferation and migration abilities were measured by MTT assay and wound healing assay.Glucose assay kit and lactic acid(LD)assay kit were used to detect changes in glycolysis levels of HGC-27 and NCI-N87 cells after treated with TAM supernatant.The biological behavior related protein expression and signaling pathway protein expression were measured by Western blot analysis.3.HGC-27 and NCI-N87 cells were blocked PI3K/mTOR or ERK pathway,and then treated with TAM supernatant.The proliferation,cell migration and glycolysis were measured.Results:1.Differentiation of TAMReal-time qPCR results showed that the expression of M2 macrophage markers CD206,CCL22 and FN1 were higher than those of the control group(P<0.05).The results of cell immunofluorescence showed that the expression of CD163 and CD206 on the surface of THP-1 cells induced with PMA and IL-4/IL-13 was significantly increased compared with the only PMA treated group.2.The effect of TAM on the proliferation,migration,glycolysis and signal pathway of gastric cancer cells.The proliferation ability of HGC-27 and NCI-N87 cells treated with TAM supernatant significantly increased(P<0.05).The cell wound healing assay data showed that the migration rate of HGC-27 and NCI-N87 cells treated by TAM supernatant was significantly up-regulated(P<0.05).The glucose consumption and lactate production of HGC-27 and NCI-N87 cells increased significantly after treated by TAM supernatant(P<0.05).Western blot results showed that the expression of p-ERK,p-AKT and p-mTOR in HGC-27/NCI-N87 cells increased after treated with TAM supernatant conditioned medium treated(P<0.05),suggesting that TAM can activate the ERK and PI3K/mTOR signal pathway.3.ERK and PI3K/mTOR signal pathways regulate the effects of TAM on the biological behavior of gastric cancer cells.After pretreatment with ERK pathway blocker(PD98059),the cell proliferation and migration rate of HGC-27 and NCI-N87 cells significantly decreased after TAM supernatant treatment.Moreover,the PI3K/mTOR dual blocker(BEZ235)can also reduce the promotion of TAM on the proliferation,migration and glycolysis of the gastric cancer cells(P<0.05).Conclusion:1.TAM can promote the proliferation,migration and glycolysis of gastric cancer cells.2.TAM can affect the proliferation and migration by regulating the PI3K/mTOR and ERK signaling pathway in gastric cancer cells.3.TAM can enhance the glycolysis of gastric cancer cells by up-regulating the PI3K/mTOR signal pathway.