Identification Of A Mesenchymal-related Signature Associated With Clinical Prognosis In Glioma

Objective Glioma mainly occurs in the brain and glial tissue and is the most common primary malignant brain tumor in adults.Among them,glioblastoma(GBM)is the worst type.GBM increases the malignancy of gliomas by enhancing cell adhesion,inflammation and the formation of new blood vessels.According to different biological,imaging and clinical characteristics,GBM is divided into 4 subtypes: proneural,neural,classical and mesenchymal.The mesenchymal subtype(MES)is a more malignant form with a higher tendency to relapse,metastasize,and increase blood vessels.In recent years,central nervous system tumors have been reclassified,which has led to a paradigm shift in personalized treatment and treatment decisions guided by prognostic factors.Even after surgery,radiotherapy,and chemotherapy,the survival time of glioma patients has only been extended by a few months.Therefore,there is an urgent need for accurate prognosis prediction and new therapeutic targets to treat gliomas.Therefore,we speculate that the detection of MES-related genes is of great significance for judging the prognosis.TCGA and CGGA,as foreign tumor databases and domestic glioma databases,provide a large number of clinical samples.Combining with matched clinical information and gene expression data,we conduct deeper research on MES-related genes as prognostic biomarkers of glioma and the development of more accurate treatment processes.Methods Use TCGA database and Ivy GAP database to obtain the intersection of MESrelated genes and get 21 genes.Obtain clinical sample information and RNA sequence information from TCGA database and CGGA database.Gene difference analysis compares TCGA glioma samples and GTEx normal samples,and 15 different genes are obtained.Univariate Cox analysis is used to determine whether 15 genes are meaningful for prognosis,and 6 of them are meaningful.R software uses Lasso regression analysis to construct risk coefficient model between each gene expression and clinical samples.According to the median risk coefficient,clinical samples were divided into high and low risk groups.Kaplan-Meier survival analysis is used to judge the difference in survival between the two groups,univariate and multivariate Cox analysis whether the risk coefficient can be used as an independent prognostic factor.The AUC curve analyzes the specificity and sensitivity of the risk factor as a prognostic factor.GSEA performs functional comparisons of related genes and predicts the functions and pathways of related genes.According to literature reports,one of the genes FCGR2 A was selected,and si RNA was used to construct a FCGR2 A silencing model,and q RTPCR was used to determine the expression of FCGR2 A and EMT marker genes in cells.Transwell migration and invasion experiments were used to detect the effects of migration and invasion of glioma cells after silenced FCGR2 A.Colony formation assay and MTT test were carried out for judge the cell proliferation ability after silenced.Westen-Blot are used to detect the expression of MES protein markers after silenced FCGR2 A.Result(1)Gene difference analysis successfully screened 15 genes that are different between the normal group and the glioma group(2)Univariate Cox analysis selected 6genes to be used as independent prognostic analysis.(3)Lasso regression analysis successfully constructed a risk coefficient model.(4)The survival analysis risk model can be used to judge the prognosis and can be used as a prognostic factor alone.(5)GSEA functional analysis revealed that MES-related genes are related to epithelialmesenchymal transition,angiogenesis,inflammation and hypoxia.(6)After silencing FCGR2 A,the ability of glioma cells to proliferate,migrate and invade decreases.(7)After silencing FCGR2 A,the expression of MES marker genes and MES protein markers were reduced.Conclusion We have determined that these 6 MES-related genes are potential predictive markers or therapeutic targets for patients with glioma.A risk model based on the expression of 6 MES-related genes was developed to better predict the OS of glioma.The functional analysis of related genes revealed that these six genes are closely related to the development of tumors,especially EMT related processes.The effect of FCGR2 A on the proliferation,migration and invasion of gliomas was studied,and it was found that after silencing the gene,the proliferation of gliomas,their migration and invasion capabilities were inhibited.Simultaneous silencing of FCGR2 A reduced the expression of MES marker genes and proteins markers,indicating that FCGR2 A may affect tumor development by affecting epithelial-mesenchymal transition.We further verified that these genes play an important role in glioma.