基于生物信息学筛选并验证上皮性卵巢癌差异microRNA及下游mRNA

Screening and verification of differential microRNA and downstream mRNA of epithelial ovarian cancer based on bioinformatics

目的:利用生物信息学对上皮性卵巢癌(EOC)的差异microRNA及其靶基因进行筛选及分析,探索EOC诊断及预后相关的潜在生物标志物。方法:本研究首先从GEO数据库(gene expression omnibus database)筛选符合条件的数据集,用limma包筛选差异microRNAs,用在线miRNet、TargetScanHuman、miRDB预测其下游靶基因,并利用WEB-based GEne SeT AnaLysis Toolkit数据库对靶基因进行GO和KEGG富集分析。最后用GEPIA2数据库对关键基因的表达与患者进展之间的关系进行分析。结果:经过筛选后,从GEO数据库获得8个microRNAs,与对照组相比,hsa-miR-4328、hsa-miR-450b-5p在EOC中表达上调,hsa-miR-3616-3p、hsa-miR-4283、hsa-miR-1208、hsa-miR-3180-3p、hsa-miR-617和hsa-miR-1207-3p在EOC中表达下调。通过3个在线网站获取差异microRNAs对应的下游靶基因,主要涉及biological regulation,membrane-enclosed lumen,protein binding等过程,主要富集于phosphatidylinositol signaling system,neurotrophin signaling pathway等信号通路。利用GEPIA2发现高表达MLLT6[HR(high)=1.3]、TIAM2[HR(high)=1.3]的EOC患者较低表达患者的生存率低(P<0.05)。还发现MLLT6、MSI1、PKMYT1、PLAGL2、ZNF514、FBXW7、GLCCI1、MFSD6、OCRL、PIP4K2B、SLC6A9与EOC疾病分期相关。通过RT-qPCR检测,发现与正常卵巢组织相比,MLLT6在EOC组织表达下调(Z=-2.602,P<0.05),其上游的hsa-miR-450b-5p在EOC组织中的表达上调(Z=-3.071,P<0.05)。结论:MLLT6可能与EOC的发生发展密切相关,且可作为提示EOC预后的指标之一;hsa-miR-450b-5p/MLLT6可能为提示EOC的生物指标。

Objective:To screen and analyze differential microRNA and its target genes in epithelial ovarian cancer(EOC)by bioinformatics,and to explore the potential biomarkers related to the diagnosis and prognosis of EOC.Methods:In this study,the eligible data sets were screened from gene expression omnibus(GEO)database,the differential microRNAs were screened by limma package,the downstream target genes were predicted by online miRNet,TargetScanHuman and miRDB,and the target genes were analyzed by GO and KEGG enrichment analysis with WEB-based GEne SeT AnaLysis Toolkit database.Then,the relationship between the expressions of key genes and patient progression were analyzed by GEPIA2 database.Results:After screening,8 microRNAs were obtained from GEO database.Compared with the control group,the expressions of hsa-miR-4328 and hsa-miR-450b-5p were up-regulated in EOC,while the expressions of hsa-miR-3616-3p,hsa-miR-4283,hsamiR-1208,hsa-miR-3180-3p,hsa-miR-617 and hsa-miR-1207-3p were down-regulated in EOC.The downstream target genes corresponding to differential microRNAs were obtained through three online websites,mainly involved in biological regulation,membrane-enclosed lumen,protein binding and other processes,mainly enriched in phosphatidylinositol signaling system,neurotrophin signaling pathway and other signal pathways.With GEPIA2,it was found that the survival rate of EOC patients with high expressions of MLLT6[HR(high)=1.3]and TIAM2[HR(high)=1.3]were lower than that of patients with low expression(P<0.05).It was also found that MLLT6,MSI1,PKMYT1,PLAGL2,ZNF514,FBXW7,GLCCI1,MFSD6,OCRL,PIP4K2B,and SLC6A9 were related to the staging of EOC disease.By real-time reverse transcription-quantitative PCR(RT-qPCR),it was found that the expression of MLLT6 in EOC tissue was down-regulated compared with that in normal ovarian tissue(Z=-2.602,P<0.05),while the expression of hsa-miR-450b-5p from upstream was up-regulated in EOC tissue(Z=-3.071,P<0.05).Conclusion:MLLT6 may be closely related to the occurrence and development of EOC and can be used as one of the indicators indicating the prognosis of EOC,and hsamiR-450b-5p/MLLT6 may be a biological indicator of EOC.