Systematic Bioinformatics Analysis Of Human TEFM And Its Expression And Clinical Significance In Brain Gliomas

Background and Objective:Glioma is a malignant tumor of the central nervous system in adults,which is mainly derived from the neuroectoderm,accounting for 49.7%of the malignant tumors in adults.Glioma is divided into two histopathological subgroups:low-grade glioma and high-grade glioma.Gliomas are relatively rare,but they can cause significant mortality and morbidity due to the complexity of the lesion site and the characteristics of the tumor itself.Although many forms of treatment are currently available,including maximal surgical resection if feasible,followed by radiation and/or chemotherapy,the median survival is relatively short.Mitochondrial transcription elongation factor(TEFM)is a protein that regulates mitochondrial gene expression and oxidative phosphorylation activity.It has been reported that the abnormal expression of TEFM gene may be closely related to the occurrence of human diseases such as mitochondrial cardiomyopathy,type I neurofibromatosis,and pancreatic cancer.However,the structural and functional studies of the TEFM gene and the proteins encoded by it are still incomplete.At the same time,the expression of TEFM in gliomas and its clinical significance have not been reported.This project uses bioinformatics methods to predict the promoter region of human TEFM gene,analyze transcriptional regulation,and analyze conservatively.The structure and function of TEFM protein were analyzed and predicted based on bioinformatics methods.The relationship between the expression level of TEFM mRNA and the clinicopathological characteristics of glioma was investigated by using databases such as TCGA and Oncomine.To investigate the correlation between the expression level of TEFM mRNA and the prognosis of patients with low-grade glioma.The expression of TEFM protein in glioma tissue and non-tumor brain tissue was detected by immunohistochemistry and tissue chip technology.This study will lay a theoretical foundation for clarifying the role of TEFM in the occurrence and development of gliomas,and also provide clues for early clinical diagnosis of gliomas and potential molecular therapeutic targets.Methods:The promoter region,transcriptional regulation function,conservation,protein physical and chemical properties,protein homology,phylogenetic tree construction,transmembrane region prediction,phosphorylation site,signal peptide site,secondary structure functional domain,tertiary structure,subcellular location,cell location,protein interaction and GSEA were predicted and analyzed by using the public database and online analysis software database.TCGA,Oncomine,COMIC,GEPIA,and cBioprotal databases were used to analyze the correlation between TEFM expression levels and pathological characteristics and prognosis of patients with glioma.28 clinical cases,51 tissue chip cases,and 10 brain trauma cases were collected to detect the expression levels of TEFM in glioma and non-tumor brain tissues using immunohistochemical techniques.Results:1.The gene promoter predicted that the human TEFM gene was located at 17q11.2,with a gene length of 8647 bp,containing 3 introns and 4 exons.The full-length transcribed mRNA of this gene is 1306 bp and encodes a protein consisting of 360 amino acids.There were at least 3 promoters in the 5’ region of the gene.The promoter region sequence contains two Cp G islands of 203 bp and 225 bp in length.There are 11 different transcription factor binding sites in the promoter region of TEFM gene.2.The systematic study of TEFM protein using different biological information methods found that the secondary structure of TEFM protein is mainly α-helix structure,and the tertiary structure is consistent with the prediction of secondary structure;the multiple sequence alignment of homologous proteins and phylogenetic tree analysis TEFM protein belongs to homologous protein.The function prediction results of TEFM protein found that:TEFM protein is localized to mitochondria,is not a membrane protein,does not have enzyme activity,and has no signal peptide site.This protein may be a transport binding protein,which plays an important role in regulating mitochondrial gene replication and transcription.Analysis of the TEFM protein interaction network and GSEA found that TEFM protein interacts with other proteins to play a role in the body,and plays an important role in mitochondrial gene expression,mitochondrial genome maintenance,DNA replication,transcription extension and nucleotide metabolism3.Analysis of Oncomine and GEPIA databases revealed that TEFM was highly expressed in gliomas.COSMIC database analysis results show that missense mutations are the main mutation types of TEFM gene in tumors.The analysis of TCGA low-grade glioma database found that the expression of TEFM mRNA is significantly correlated with age,WHO grade,pathological types,headache history and supratentorial location in glioma patients.Kaplan-Meier analysis showed that a high expression level of TEFM mRNA indicated a poor prognosis in OS of low-grade glioma patients.Multivariate Cox regression analysis showed that age,WHO grade,pathological types and supratentorial location were the independent prognostic factors of patients with glioma4.The cBioprotal database was used to analyze the correlation between TEFM genes and the top ten genes,mitochondrial regulatory genes and glioma marker genes in low-grade glioma.Among the top ten genes co-expressed with TEFM genes,the expression levels of UTP6,ZNF700,MAGOHB,ZNF140,NUF43,PIGW,RFC4 and NOL11 genes were positively correlated with TEFM gene expression levels,while the expression levels of KIF1A and CLSTN1 genes were negatively correlated with TEFM gene expression levels.Among mitochondrial transcriptional regulatory genes,the expression levels of TFB1M,TFB2M,MTERF1,MTERF2,MTERF3,POLMTR and NRF1 genes were positively correlated with TEFM gene expression levels Among glioma marker genes,IDH1,TERT,ATRX,MKI67 and TP53 gene expression levels were positively correlated with TEFM gene expression levels,while MGMT and BRAF gene expression levels were negatively correlated with TEFM gene expression levels5.GSEA results show that cell cycle pathways,DNA replication pathways,RNA degradation pathways,ubiquitin-mediated proteolysis pathways,aminoacyl-tRNA biosynthetic pathways,pyrimidine metabolic pathways,N-glycan biosynthetic pathways,nucleotide excision repair pathway and spliceosome pathway were significantly associated with TEFM expression in the TCGA glioma data set6.Immunohistochemistry and tissue chip analysis revealed that the expression of TEFM protein in glioma tissue was significantly higher than that in normal brain tissue.Tissue chip technology was used to analyze the expression level of TEFM relative expression in various pathological parameter groups in gliomas.The results showed that the relative expression of TEFM was up-regulated or down-regulated in the pathological type,Ki67 expression level and IDH1 mutation group(P<0.05).In tissue microarray,the correlation analysis between the relative expression level of TEFM and related pathological parameters of glioma patients found that the relative expression of TEFM protein was significantly related to the IDH1 mutations of glioma patients.The Kaplan-Meier prognosis model was used to analyze the pathological parameters and relative expression levels of TEFM in glioma patients and the prognosis of glioma patients.The results showed that there was a significant correlation between WHO grade and OSConclusion:1.Predictive analysis of TEFM gene promoter and transcription factor binding sites using bioinformatics methods,and predictive analysis of TEFM protein structure and function.This provides a bioinformatics reference and theoretical basis for further research on the functional research of TEFM protein.At the same time,it also lays a foundation for further exploring the mechanism of action of TEFM gene in the genesis and development of malignant tumors,and the search for new tumor gene therapy targets2.Based on the analysis of TCGA,Oncomine,GEPIA,and cBioprotal databases,it was found that the expression level of TEFM in low-grade glioma was increased and related to the pathological characteristics and prognosis of glioma patients,suggesting that TEFM may be involved in the development and occurence of low-grade glioma It has laid a foundation for further research on the role of TEFM in the occurrence and development of low-grade glioma.The results of GSEA enrichment analysis predicted the signal pathway of the role of TEFM in gliomas,which laid the foundation for further research on the mechanism of TEFM in low-grade glioma.It also provides potential molecular targets for finding new markers of glioma and treating gliomas3.Immunohistochemistry and tissue chip technology detected that TEFM protein was highly expressed in glioma tissue.Tissue microarray analysis showed that the relative expression of TEFM was up-regulated or down-regulated in the pathological type,Ki67 expression level and IDH1 mutation group,and the relative expression level of TEFM protein was significantly correlated with the presence or absence of IDH1 mutation.It is suggested that TEFM may be involved in the occurrence and development of gliomas,and may be closely related to the malignancy of gliomas.