miR⁃135b⁃5p在口腔鳞状细胞癌中的表达及相关生物信息学分析

Expression and relevant bioinformatics analysis of miR⁃135b⁃5p in oral squamous cell carcinoma

目的探索miR⁃135b⁃5p在口腔鳞状细胞癌(oral squamouscellcarcinoma,OSCC)及癌旁组织中的表达及其临床意义,预测miR⁃135b⁃5p靶基因并进行相关生物信息学分析。方法通过肿瘤与癌症基因图谱(the CancerGenomeAtlas,TCGA)、基因表达数据库(Gene ExpressionOmnibus,GEO)数据库分析miR⁃135b⁃5p在OSCC组织和癌旁组织中的表达并分析其临床意义;临床收集新鲜组织标本,采用实时荧光定量PCR验证不同组织中miR⁃135b⁃5p的表达情况。采用生物信息学方法预测miR⁃135b⁃5p的靶基因并进行通路富集分析。构建蛋白互作网络筛选关键靶基因。结果OSCC组织中miR⁃135b⁃5p表达量较癌旁组织上调,差异有统计学意义(P<0.001);miR⁃135b⁃5p表达量对OSCC组织具有良好的诊断效能(AUC=0.960,P<0.001);OSCC组织中miR⁃135b⁃5p表达水平与组织病理分级相关(P=0.011);生信分析结果显示,miR⁃135b⁃5p的靶基因富集在与肿瘤相关的钙离子、cGMP⁃PKG、cAMP信号通路中;筛选得到10个关键靶基因:DLG2、ANK3、ERBB4、SCN2B、NBEA、GABRB2、ATP2B2、SNTA1、CACNA1D、SPTBN4。结论miR⁃135b⁃5p可作为一种促癌基因参与OSCC的发生发展,并具有成为OSCC诊断标志物及治疗靶点的潜在应用价值。

Objective To observe the clinical significance of miR⁃135b⁃5p in oral squamous cell carcinoma(OSCC)tissues and to conduct a bioinformatics analysis of its predicted target genes.Methods The expression levels of miR⁃135b⁃5p in OSCC tissues and adjacent normal tissues were compared using data from TCGA and GEO databases,and the correlations of miR⁃135b⁃5p expression level with clinicopathologic characteristics were analyzed.Fresh tissues were collected in the clinic,and the expression of miR⁃135b⁃5p was verified by quantitative real⁃time PCR.The target genes with enriched pathways were analyzed by using bioinformatics methods.A protein⁃protein interaction network was constructed to screen hub genes.Results The expression levels of miR⁃135b⁃5p were significantly upregulated in OS⁃CC tissues compared to adjacent normal tissues(P<0.001)and had a good diagnostic capability(AUC=0.960,P<0.001).The expression level of miR⁃135b⁃5p was positively correlated with histopathological grading(P=0.011).En⁃richment analyses revealed that the target genes of miR⁃135b⁃5p were significantly associated with tumor⁃related signaling pathways,such as the calcium signaling pathway,the cGMP⁃PKG signaling pathway and the cAMP signaling path⁃way.Ten core target genes were obtained by screening:DLG2,ANK3,ERBB4,SCN2B,NBEA,GABRB2,ATP2B2,SN⁃TA1,CACNA1D,and SPTBN4.Conclusion miR⁃135b⁃5p may act as an oncogene miRNA in OSCC and has the po⁃tential value of acting as a diagnostic biomarker and therapeutic target for OSCC.