Analysis of key pathogenic target genes of ovarian cancer and experimental verification of cells in vitro

Objective:To mine genes highly related to the pathogenesis of ovarian cancer by using multichip integrated bioinformatics methods and verify them in cells,which provided key genes and important theoretical basis for targeted research of ovarian cancer.Methods:Three datasets,GSE38666,GSE40595 and GSE54388,were downloaded from the Gene Expression Integrated Database database for differential gene(DEGs)screening,including 26 normal samples and 65 ovarian cancer samples.Gene ontology functional annotation of selected DEGs was performed through DAVID online database to clarify the biological characteristics of DEGs.The main pathways of DEGs were obtained by enrichment analysis using Kyoto gene and genomic encyclopedia method.Based on the STRING database,the DEGs protein-protein interaction network was constructed by using CytoScape software,and the key genes were screened by GEPIA2 database to verify the expression at the cell level.Results:A total of 238 DEGs were screened from GSE38666,GSE40595 and GSE54388 datasets,of which 168 DEGs were upregulated and 70 DEGs were down-regulated.The co-expressed DEGs were mainly enriched in biological functions such as mitotic nuclear division,spindle,chromosomal region and DNA helicase activity in ovarian cancer.They were mainly involved in biological processes such as cell cycle,DNA replication,oxidative phosphorylation and biosynthesis of amino acids,thereby affecting the occurrence and development of ovarian cancer.Six genes were highly expressed and associated with the development of ovarian cancer,including IFI27,EPCAM,CXCR4,PEA15,CLDN3 and CAPG.Cell verification showed that the mRNA expression of the six genes in ovarian cancer cells was higher than that in normal ovarian cells(P<0.05),which was consistent with the previous screening results.Conclusion:Multi-chip integrated bioinformatics is an effective method to find ovarian cancer target genes.IFI27,EPCAM,CXCR4,PEA15,CLDN3 and CAPG are highly correlated with the occurrence and development of ovarian cancer,which can be used as target genes for ovarian cancer research.

卵巢癌致病关键靶基因分析及体外细胞实验验证

目的:通过多芯片整合的生物信息学方法挖掘与卵巢癌发病高度关联的靶基因,并通过体外细胞实验进行验证,为卵巢癌的靶向研究提供关键基因及重要理论依据。方法:(1)对基因表达综合数据库中与卵巢癌相关数据集GSE38666、GSE40595和GSE54388进行差异基因(DEGs)筛选,其中包括26个正常样本和65个卵巢癌样本。(2)通过DAVID在线数据库对筛选的DEGs进行基因本体功能注释,明确DEGs的生物学属性特征。(3)通过京都基因与基因组百科进行富集分析,获得DEGs的主要作用通路。(4)基于STRING数据库运用CytoScape软件完成DEGs蛋白质-蛋白质相互作用网络的构建,并结合GEPIA 2等数据库筛选出关键基因。(5)采用qRT-PCR对卵巢癌细胞关键基因的表达进行验证。结果:从GSE38666、GSE40595和GSE54388 3个数据集中筛选出238个共同表达的DEGs,其中168个表达上调,70个表达下调,在卵巢癌中主要富集于有丝分裂核分裂、纺锤体、染色体区域和DNA解旋酶,主要参与细胞周期、DNA复制、氧化磷酸化和氨基酸的生物合成等生物过程,影响卵巢癌的发生、发展。其中IFI27、EPCAM、CXCR4、PEA15、CLDN3和CAPG 6个差异表达基因与卵巢癌高度相关,体外细胞实验结果与其一致。结论:多芯片整合的生物信息学方法是筛选卵巢癌靶基因的有效方法,IFI27、EPCAM、CXCR4、PEA15、CLDN3和CAPG与卵巢癌发生、发展高度相关,有望成为卵巢癌致病关键靶基因,需要进行深入研究。