人食管鳞状细胞癌中miR-424的差异表达及生物信息学分析

The differential expression and bioinformatics analysis of miR-424 in human esophageal squamous cell carcinoma

目的:探讨hsa-miR-424(miR-424)在人食管鳞状细胞癌发生发展机制中的作用,并对其生物学特征及功能进行预测总结分析,为最终研究miR-424的功能提供有利的线索。方法:通过实时荧光定量PCR方法,检测食管鳞状细胞癌患者石蜡标本和癌旁组织,以及人食管鳞状细胞癌细胞株kyse520、kyse410和正常人食管上皮细胞系Het-1A中miR-424相对表达水平。应用MIRDB、miRanda、Target Scan三种在线工具预测miR-424的靶基因,取三者预测结果的交集,并结合DIANALAB-TarBase7.0数据库和miRTarBase数据库两个已经实验证实的基因数据库,确定miR-424的靶基因范围。最后通过使用在线数据库metascape对has-miR-424预测的靶基因集合进行基于GO、KEGG数据库的功能注释分类和通路富集分析。结果:miR-424在食管癌细胞系kyse520和kyse410中的表达显著高于正常食管上皮细胞系Het-1A的表达(P<0.01)。与对应的癌旁组织相比较,鳞癌组织中miR-424的表达均显著增高,其中,约77.8%(56/72)的患者癌组织中miR-424的表达较癌旁组织上调了超过50%。miR-424在物种进化上具有高度保守性,其靶基因功能主要集中在调控细胞生长、增殖和正负向调控细胞凋亡;参与蛋白质脱磷酸和泛素化,组蛋白修饰和甲基化,DNA损伤修复等生物学过程(P<0.01)。主要参与的信号通路有p53信号通路、PI3K-Akt信号通路、MAPK信号通路和酪氨酸激酶信号通路(P<0.01)。结论:miR-424在食管鳞状细胞癌中是高表达状态,提示miR-424可以作为癌症促进子参与食管癌的发生发展。

To explore the hsa-miR-424 expression in the cell lines of human esophageal squamous cell carcinoma,predict and analyze the target genes of hsa-miR-424 using bioinformatic method and to provide theoretical guidance for the further function study.Methods:The relative expressions of hsa-miR-424 were detected by real-time quantitative PCR in human esophageal squamous cell carcinoma cell lines kyse520,kyse410 and human normal esophageal epithelial cell line Het-1A,and esophageal squamous cell carcinoma tissues and corresponding nomal esophageal tissues.hsa-miR-424 target genes were predicted by MIRDB,miRanda and Target Scan,then combined with the confirmed gene database DIANALAB-TarBase7.0 and miRTarBase.Functional annotation classification and pathway enrichment analysis based on GO and KEGG databases were performed on target gene sets predicted by miR-424 using the online database metascape.Results:Compared with normal esophageal epithelial cells,the expression levels of hsa-miR-424 in human esophageal squamous cell carcinoma cell lines kyse520,kyse410 were significantly higher.Compared with normal esophageal epithelial tissues,the expression levels of hsa-miR-424 in human esophageal squamous cell carcinoma tissues were significantly higher.About 77.8 percent(56/72)of the patients had a upregulation of more than 50 percent.hsa-miR-424 is highly conserved in species evolution.The function of the target genes mainly focused on regulating cell growth,proliferation and positive and negative regulation of apoptotic process,and participating in the biological processes of protein dephosphorylation and ubiquitination,histone modification and methylation,regulation of response to wounding(P<0.01).hsa-miR-424 target genes were significantly enriched in the MAPK,p53 and tyrosine kinase signal pathway closely related with tumor(P<0.01).Conclusion:The highly expression of hsa-miR-424 in esophageal squamous cell carcinoma suggesting that hsa-miR-424 may be involved in the regulation of pathogenic of esophageal squamous cell carcinoma.