Comprehensive Analysis of circRNA-associated Competing Endogenous RNA Networks and Immune Infiltration in Gastric Cancer
crossref(2021)
Abstract
Abstract BackgroundCircular RNA (circRNA) has been shown to be an important regulator in gastric cancer (GC). However, functions and regulatory mechanisms of circRNA-related competitive endogenous RNA (ceRNA) in GC have not been established.MethodsCircRNA data and clinical data were downloaded from the GEO and TCGA databases. The ceRNA and Protein-Protein Interaction (PPI) networks were constructed through bioinformatics analysis. Function enrichment analysis was performed. Additionally, correlations between expression levels of the top 10 hub genes and immune cell infiltration levels, histopathological grade and clinical stage were determined to establish their clinical values. The differentially expressed circRNA (DEcircRNA) was validated by quantitative real-time PCR (qRT-PCR).ResultsScreening of the GEO and TCGA databases revealed a total of 1627 DEcircRNAs, 6516 DEmRNAs, and 1451 DEmiRNAs. The ceRNA interaction network contained 2 circRNAs, 3 miRNAs and 55 mRNAs. Meanwhile, Gene Ontology (GO) analysis revealed a total of 323 biological processes (BP) terms, 53 cellular components (CC) terms, 51 molecular functions (MF) terms, while the Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed 4 signaling pathways. Gene Set Enrichment Analysis (GSEA) analysis revealed that EPHA4, NCAM1 and NRXN1 were positively correlated with the axon guidan and adhesion molecules pathways. Most of top 10 hub genes were positively correlated with B cells, CD8+ T cells, CD4+ T cells, Neutrophils and Dendritic Cell infiltration. Correlation analysis between hub genes and clinical phenotypes revealed that elevated expressions of EPHA4 and KCNA1 indicated poor tissue differentiation and were associated with clinically advanced stages of GC. The qRT-PCR results revealed that the expression of has_circ_0002504 was significantly down-regulated in 3 GC cell lines which was consistent with the results of our bioinformatics analysis.ConclusionsHas_circ_0001998 and has_circ_0002504 are potential diagnostic biomarkers for GC, and the high expressions of both EPHA4 and KCNA1 may predict poor prognosis.
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Key words
endogenous circrna-associated networks,gastric cancer,immune infiltration
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