基于生物信息学分析探讨circRNA在口腔鳞状细胞癌中的调控作用

Exploring the Regulatory Role of circRNA in Oral Squamous Cell Carcinoma Based on Bioinformatic Analysis

目的通过生物信息学方法和qRT-PCR实验筛选并验证口腔鳞状细胞癌(OSCC)中差异表达的环状RNA(circRNA),并构建OSCC中可能的circRNA-miRNA-mRNA调控网络。方法选取GEO数据库中2个OSCC的circRNA高通量数据,使用R软件识别OSCC中的差异circRNA,对筛选出的circRNA取交集后获得候选circRNA。通过qRT-PCR在人口腔黏膜角质细胞系(HOK)和OSCC细胞系中验证候选circRNA的表达水平。使用circInteractome和circBank数据库预测circRNA下游的miRNA,通过miRDB和TargetScan数据库预测miRNA的靶基因。使用DAVID在线数据库对靶基因进行GO功能和KEGG通路富集分析。通过STRING数据库和Cytoscape软件确定核心基因,采用GEPIA生存分析网站分析其与预后的关系。结果共筛选出3个差异circRNA、7个miRNA和2575个潜在靶mRNA。GO和KEGG富集分析表明,靶基因主要富集在肿瘤相关的通路和生物学过程。基于富集分析、蛋白质相互作用网络图构建和核心基因的生存分析结果,筛选出3个核心基因(EGF、STAT 5 A和LEF 1),从而建立OSCC相关的circRNA-miRNA-核心基因调控网络。结论本研究为circRNA在OSCC中的调控机制提供参考,为OSCC的诊断及靶向治疗拓宽思路。

Objective To identify differentially circRNA in oral squamous cell carcinoma(OSCC)using bioinformatics and qRT-PCR,and to investigate the role of the circRNA-miRNA-mRNA regulatory network in OSCC.Methods Two circRNA expression datasets of OSCC were acquired from the gene expression omnibus(GEO)database.Then,differentially expressed circRNA in OSCC were screened using R software.The candidate circRNA were obtained by intersecting the screened circRNA.The expression levels of the candidate circRNA were validated in human oral keratinocyte(HOK)and OSCC cell lines by qRT-PCR.The miRNA that bind to differentially expressed circRNA were predicted with circInteractome and circBank software.Target mRNA prediction of miRNA was performed by using miRDB and TargetScan software.The target genes were subjected to GO function and KEGG enrichment analysis by DAVID.The STRING database and Cytoscape software identified the hub genes,and their relationship with prognosis was analyzed using the GEPIA survival analysis website.Results Our data identified 3 differentially expressed circRNA,7 miRNA,and 2575 potential target mRNA.GO and KEGG analysis demonstrated that the target genes were potentially implicated in OSCC pathogenesis.Three hub genes(EGF,STAT5A,LEF1)were screened out based on enrichment analysis,protein interaction network construction,and the results of survival analysis of hub genes.The the OSCC-related circRNA-miRNA-hub genes regulatory network was constructed.Conclusions Our study provided reference and broadened the thinking for the study of circRNA in the pathogenesis and treatment of OSCC.