Regulation Of Coleractal Cancer Cells Expressed High Level Of MTA1 To Tumor Associated Macrophages In Tumor Microenvironment Of Coleractal Cancer

Regulation of coleractal cancer cells expressed high levels of MTA1 totumor associated macrophages in tumor microenvironment of Coleractal cancerObjective Metastasis associated protein 1(MTA1)is upregulated in a variety of human tumor cells.As a subunit of ATP-dependent chromatin remodeling complex,MTA1 can extensively affect downstream gene expression.Therefore,it has a regulatory effect on cytokines,chemokines and tumor microenvironment secreted by tumor cells.However,the regulation of MTA1 overexpression in tumor cells on the immune state of the local microenvironment is still.unknown.The aim of this study was to elucidate the role of MTA1 overexpression in the regulation of macrophage infiltration and functional activation in the tumor microenvironment and the anti-tumor killing effect of CTL based on colon cancer species.Methods TCGA colorectal cancer cohort transcriptome data and TIMER immune infiltration database were analyzed to evaluate the relationship between MTA1 expression and chemokine expression and immune cell infiltration,and the regulation of MTA1 high expression on chemokine expression was detected using cytokine antibody microarray.The expression levels of inflammation-related molecules in macrophages were detected by qPCR.The polarization surface marker molecules of macrophages,autophagy level,expression of molecules related to functional activation and functional depletion of lymphocytes,apoptosis level of lymphocytes,autophagy level,apoptosis level of tumor cells,interaction between tumor cells,macrophages and lymphocytes were detected by flow cytometry.The expression levels of CD8,CD4,CD20,CD206,CD163,CD68,CD86,PD-L1 and MTA1 in colorectal cancer tissues were detected by multiple fluorescence immunohistochemistry assay.Inform2.2.4 was used for image analysis,R package ISAT was used for distance analysis,Flow Jo software was used for Flow cytometry test results analysis,and Graphpad8.3 was used for all data statistical analysis.Results In the assessment of the expression level of MTA1 in tumor cells and its chemotactic efficacy,the high expression of MTA1 in colon cancer cells significantly down-regulated the expression level of chemokines related to monocyte recruitment and up-regulated the cytokines related to T cell recruitment.In terms of the induction of macrophage polarization,the polarization level of M2 macrophages characterized by CD206 positive was increased,and the polarization level of M2 macrophages characterized by CD 163 positive was decreased,but the induction of M1 macrophages was not significantly down-regulated.In the co-culture system with high expression level of MTA1,the anti-inflammatory cytokine IL-10 and pro-inflammatory cytokines iNOS,TNF-α and IL-6 secreted by macrophages were significantly increased compared with the negative control.In the co-culture system with high expression of MTA1,the secretion of pro-inflammatory cytokine IL-6 from macrophages was significantly lower than that in the two groups with low expression of MTA1.In the co-culture system with high expression of MTA1,the activation,depletion and apoptosis levels of lymphocytes were significantly higher than those in the control group and MTA1 knockdown group.However,the interaction between tumor cells and lymphocytes in the MTA1 high expression group wassignificantly lower than that in the control group and MTA1 knockdown group.The process of increased autophagy level after TCR activation was also significantly lower than that of the control group and MTA1 knockdown group.In the end,the killing effect of co-culture system with high MTA1 expression level was significantly lower than that of co-culture system with low MTA1 expression level.At the same time,flow cytometry showed that the presence of macrophages in the co-culture system could not only reduce the expression of activation marker molecules of lymphocytes and increase the expression level of functional depletion molecules,but also reduce the apoptotic level of lymphocytes,thus greatly improving the killing efficacy of lymphocytes against tumor cells.Conclusions MTA1 high expression in colorectal cancer caused by macrophages in tumor microenvironment of raise reduction,reduced secretion of macrophage inflammatory factors and antigen presenting process is missing.then the T cells antitumor effects cannot be activated,inhibition ratio increases,resulting in local immune suppression in the-tumormicroenvironment,immune escape of tumor cells.Analysis of the relationship between immune cell subsets and cancer progression in colorectal cancermicroenvironment based on Single-cell transcriptome sequencing dataObjective To explore the correlation between the components of immune cell subsets in tumor microenvironment of colorectal cancer and the progression of colorectal cancer disease,and to explore the differential regulational effects of different CD4+T cell subsets on the progression of colorectal cancer at the level of regulational function,based on the high-resolutional single cell transcriptiome sequencing data.Methods Using the single cell transcriptiome sequencing data of colorectal cancer patients in GEO database,CibersortX algorithm was used to assess the immune cell subsets score from the transcriptiome sequencing datas of colorectal cancer patients in the Cancer Genome Atlas(TCGA)database.The obtained immune cell scores were combined with clinical data to analyze the correlation between metastasis and immune cell.subsets related genes were calculated based on immune cell scores,and functional enrichment scoresperformed by using David analysis platform were used to compare the different regulational function of immune cell subsets in colorectal cancer.Lasso algorithm was used to screen immune cell subsets and Cox analysis was used to construct a risk prognosis model.Results The abundance of CD4-TCF7 was significantly increased in metastatic patients of colorectal cancer compared with non-metastatic patients(P<0.01),the abundance of CD4CTLA4,CD8-LEF1,PLASMAB-IgG,and MACRO-IL1B decreased in metastatic patients(p<0.05).In the survival analysis,15 of the 38 immune cell subsets were significantly associated with over survival based on the AUC curve(P<0.05).After analysing the regulational functions of CD4+T cell subsets in colorectal cancer cell,we find the CD4-GNLY,CD4-ANXA1 and CD4-CXCR6 with more powerful regulation on the colorectal cancer angiogenesis related pathways compare to others,CD4-IL23R preferentially enriched in energy metabolism related pathways,CD4-GZMK comparedto other subsets uniquely enriched in hypoxia stress related pathways,CD4-ANXA1,CD4-TCF7 and CD4-CTLA4 compared to other subsets preferentially enriched in the cell proliferation related pathways,CD4-ANXA1,CD4-TCF7,CD4-CTLA4,and CD4CXCL13 preferentially enriched in the pathways of cell adhesion and cell migration compared to other subsets.Based on the composition of immune cell subsets and the immune state represented by them,the prognosis evaluation of patients has a good effect and a certain practical valueConclusion There are significant differences in the composition of immune cell subsets in tumor microenvironment of colorectal cancer between metastatic and non-metastatic patients,and there are differences in the regulational functions of immune cell subsets regulating colorectal cancer cell functions.The composition of immune cell subsets in colorectal cancer immune microenvironment affects the survival and prognosis of patients.