摘要
目的筛选食管鳞癌(ESCC)组织中差异表达的miR-17-92基因簇及其靶基因,分析靶基因的生物学功能,并探讨miR-17-92基因簇及其靶基因与患者临床病理特征、预后的关系。方法下载癌症基因组图谱(TCGA)数据库和基因表达综合(GEO)数据库(GSE43732数据集)中ESCC相关数据,分别采用edgeR包和GEO2R分析miR-17-92基因簇的表达情况。将ESCC组织、正常食管组织及癌旁组织中差异表达最显著(差异倍数>2倍,校正后P<0.05)的miR-17-92基因簇成员作为关键miRNAs。采用miRDB 6.0、TargetScan 7.2及miRTarBase 8.0在线网站预测关键miRNAs的靶基因,经蛋白互作网络分析及差异表达分析确定关键靶基因。对关键靶基因进行KEGG信号通路和GO功能富集分析。采用独立样本t检验或Mann-Whitney U检验分析关键miRNAs及其关键靶基因与ESCC临床病理学特征间的关系,采用生存分析评价关键miRNAs及其关键靶基因与ESCC患者预后的关系。结果相对于正常食管组织及癌旁组织,ESCC组织中miR-17-92基因簇表达均上调,其中上调的关键miRNAs是miR-17-5p、miR-18a-5p。筛选出14个差异表达的关键靶基因,分别为CNOT6L、CTDSPL、ATM、FBXL5、NHLRC3、FEM1C、HIF1A、UBE2G1、CENPQ、CNOT7、RHOC、NR3C1、RUNX1、E2F3。关键靶基因主要富集于信号传导、转录调控、癌症通路、细胞周期等肿瘤相关生物学过程或通路。miR-17-5p、miR-18a-5p和关键靶基因CTDSPL在不同ESCC分化程度间存在差异表达(P均<0.05)。关键靶基因ATM和NHLRC在ESCC不同T分期间存在差异表达(P均<0.05)。关键靶基因UBE2G1在ESCC不同N分期间存在差异表达(P=0.03)。生存分析结果表明,关键靶基因ATM的表达水平与ESCC患者的预后相关(P<0.01)。结论 ESCC组织中miR-17-92基因簇表达上调,关键miRNAs为miR-17-5p、miR-18a-5p,关键靶基因为CNOT6L、CTDSPL、ATM、UBE2G1等14个。关键靶基因主要富集于信号传导、转录调控、癌症通路等肿瘤相关生物学过程或通路。CTDSPL、ATM、NHLRC、UBE2G1与ESCC患者临床病理特征有关,ATM的低表达与ESCC患者的预后差有关。
Objective cell carcinoma(ESCC),we selected the differentially expressed miR-17-92 cluster members and their target genes in ESCC tissues,analyzed the biological functions of the target genes,and explored the relationship between the differentially expressed miR-17-92 cluster members and their target genes with the clinicopathological characteristics and prognosis of patients.Methods(TCGA)database and downloaded GSE43732 of 119 pairs of ESCC samples from gene expression omnibus(GEO). Differential expression of miR-17-92 cluster between normal and tumor samples were identified by using edgeR package and GEO2R tool. The miR-17-92 cluster members with the most significant differential expression in ESCC tissues(fold change>2 and adjusted P<0. 05 as cut-off criteria)were defined as key miRNAs. MiRDB 6. 0,TargetScan 7. 2 and miRTarBase 8. 0 were used to predict the target genes of key miRNAs,and the key target genes were determined by protein interaction network analysis and differential expression analysis. The GO functional enrichment and KEGG signaling pathway analysis of key target genes were carried out. Independent sample t test or Mann-Whitney U test was used to analyze the relationship between key miRNAs and their key target genes and the clinicopathological characteristics of ESCC. Survival analysis was used to evaluate the correlation between key miRNAs and their key target genes and the prognosis of ESCC patients.Results up-regulated expression in ESCC tissues,and miR-17-5p and miR-18a-5p were the most differentially expressed miRNAs which were defined as key miRNAs. Fourteen key target genes with differential expression in ESCC tissues were screened out,namely CNOT6 L,CTDSPL,ATM,FBXL5,NHLRC3,FEM1C,HIF1A,UBE2G1,CENPQ,CNOT7,RHOC,NR3 C1,RUNX1,and E2 F3. Gene functional enrichment analysis found that key target genes were mainly enriched in tumor-related biological processes and pathways like signal transduction,transcription,pathways in cancer,and cell cycle pathway,etc. The miR-17-5p,miR-18a-5p and the key target gene CTDSPL were differentially expressed in different levels of ESCC differentiation(P=0. 02,P=0. 04 and P<0. 01,respectively). The key target genes ATM and NHLRC were differentially expressed in different T scores of ESCC tissues(P=0. 01 and P=0. 03,respectively). The key target gene UBE2G1 was differentially expressed in different N scores of ESCC tissues(P=0. 03). The expression level of the key target gene ATM was related to the prognosis of ESCC patients(P<0. 01).Conclusions ter was up-regulated in ESCC tissues,miR-17-5p and miR-18a-5p were defined as key miRNAs,and 14 target genes such as CNOT6 L,CTDSPL,ATM,UBE2G1,etc. were regarded as key target genes. CTDSPL,ATM,NHLRC,and UBE2G1 were related to the clinicopathological characteristics of ESCC patients,and low expression of ATM was associated with poor prognosis of ESCC patients.
作者
邵毅
沈艺
牛琛
阮晓莉
Rena Nakyeyune
刘芬
SHAO Yi;SHEN Yi;NIU Chen;RUAN Xiaoli;RENA Nakyeyune;LIU Fen(Capital Medical University,Beijing Key Laboratory of Clinical Epidemiology,Beijing 100069,China)
出处
《山东医药》
CAS
2021年第15期1-5,共5页
Shandong Medical Journal
基金
北京市教委科研计划一般项目(KM201710025006)
国家自然科学基金资助项目(81874277)。
作者简介
第一作者:邵毅(1995-),女,硕士,主要从事肿瘤流行病学和卫生统计的研究工作。E-mail:shaoyi200812@sina.com;通信作者:刘芬(1974-),女,教授,博士生导师,主要从事肿瘤流行病学和卫生统计的研究工作。E-mail:liufen05@ccmu.edu.cn。
