| Objective: To screen differentially expressed lnc RNAs using transcriptome sequencing technology and bioinformatics analysis,predict their role in oral squamous cell carcinoma and validate their expression levels in oral squamous carcinoma tissues and cells.METHODS: In this study,cancer tissues and normal tissues adjacent to cancer from three pairs of oral squamous carcinoma patients were collected and sequenced using transcriptome high-throughput sequencing technology to construct libraries of differential expression profiles of lnc RNAs,m RNAs and mi RNAs.The OSCC expression data containing 33547 genes were also downloaded from the GEO database,and the sequencing data were combined with the database data to screen differentially expressed lnc RNAs as candidate genes.The ce RNA network was constructed,and the functions and pathways of the ce RNA network were analyzed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis methods.OSCC expression data were downloaded from the TCGA database to analyze the expression levels of candidate genes and their localization in the cells.The role of candidate genes in OSCC immune infiltration and their correlation with tumor TNM stage,etc.were analyzed using R software.Primers were designed for the screening genes,and their expression levels were verified in 40 pairs of OSCC tissues and their paraneoplastic normal tissues and HOK,SCC25 and CAL27 cell lines.RESULTS: In this study,968 differentially expressed lnc RNAs were identified by transcriptome sequencing of 3 pairs of OSCC tissues and paired paraneoplastic tissues.11,674 genes were obtained by downloading the GSE30784 dataset from the GEO database,and a total of 75 differentially expressed lnc RNAs were obtained by combining them with sequencing data for analysis.the highest differentially expressed ploidy was identified The results of GO and KEGG showed that GO results were mainly enriched in organelle assembly and transcriptional co-regulator activity,while KEGG was enriched in transcriptional dysregulation and central carbon metabolism pathways in cancer,suggesting that it may affect tumor development by regulating transcriptional processes.The results of TCGA database validation showed that LINC01560 expression was elevated in oral squamous cell carcinoma and was significantly expressed in salivary glands,mainly in the nucleus,nucleoplasm and cytoskeleton of the cells.Immune infiltration correlation analysis showed that the expression of LINC01560 correlated with the immune cell levels of B cells,T cells CD4,neutrophils and dendritic cells,suggesting that it may be involved in regulating the immune infiltration process of OSCC.Meanwhile,LINC01560 may correlate with OSCC cancer N-stage and tumor grade.Expression validation results showed that LINC01560 was highly expressed in both OSCC tissues and cells,which was consistent with the sequencing and database results.The expression of LINC01560 was statistically analyzed using ROC curves,and the P value of LINC01560(AUC=0.709,P=0.008)was <0.05,and the area under the AUC curve was >0.7.Conclusions: 1.LINC01560 was significantly associated with OSCC as determined by screening whole transcriptome sequencing data with GEO database data cross-analysis;2.LINC01560 expression in OSCC tissues and cells was higher than that in normal oral tissues,consistent with the database validation results,and its ROC curve AUC > 0.7,suggesting that LINC01560 may serve as a clinical LINC01560 and its ce RNA may influence the development of OSCC by regulating the transcriptional process;its expression correlates with the immune cell levels of B cells,T cells CD4,neutrophils and dendritic cells,suggesting that it may be involved in regulating the immune infiltration of OSCC;meanwhile,the abnormal expression of LINC01560 may be associated with OSCC cancer N stage and tumor grade. |
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