摘要
目的应用生物信息数据库分析和分子生物学方法检测乙型肝炎病毒(HBV)对微小RNA(miR-10b)表达的影响,以及生物信息学方法探讨HBV影响的miR-10b异常表达在肝脏疾病中的作用。方法以GEO2R分析miRNA表达芯片GSE19980中miR-10b的表达水平,荧光定量聚合酶链反应(PCR)验证分析结果。Targetscan、miRTarBase、miRDB和DIANA TOOLS分析miR-10b的靶基因并用韦恩图进行筛选。应用DAVID数据库对miR-10b靶基因数据集进行基因本体(GO)功能富集分析。采用STRING分析软件进行KEGG信号通路分析。GEO2R分析HBV诱导的肝细胞癌及其邻近正常组织的差异表达基因,并与miR-10b靶基因进行韦恩图筛选以获得交集靶基因,然后用STRING分析蛋白质的相互作用,得到miR-10b靶基因的核心蛋白基因。结果miR-10b在稳定表达HBV的HepG2.2.15细胞中表达明显上调,差异有统计学意义(P<0.05);miR-10b靶基因主要富集在转录调控、转录因子活性、参与转录因子复合体等相关蛋白基因;miR-10b的靶基因富集在蛋白多糖、miRNA、cAMP信号通路;miR-10b靶基因编码蛋白的相互关系分析获得CREB1、NCOA6、NCOR2、PIK3CA、GATA3、MAPRE1、TIAM1等核心蛋白,其功能涉及糖稳态调节、炎症、肿瘤发生和发展。结论HBV上调miR-10b表达,高表达的miR-10b对其靶基因的沉默可能在HBV感染的肝脏疾病中发挥着重要作用。
Objective To detect the effect of hepatitis B virus(HBV)on miR-10b expression by the bioinformatics database analysis and molecular biology method,and to explore the role of HBV influenced miR-10b aberrant expression in liver diseases by using the bioinformatics method.Methods The miR-10b expression level in the miRNA expression chip GSE19980 was analyzed by GEO2R,and the analytic results were verified by fluorescence quantitative PCR.The target genes of miR-10b were analyzed by using Targetscan,miRTarBase,miRDB and DIANA TOOLS,and screened by Venn diagram.The miR-10b target gene data set conducted the gene ontology(GO)functional enrichment analysis by using the DAVID database.The KEGG signal pathway analysis was performed by using STRING analysis software.The HBV-induced differentially expressed genes of the hepatocellular carcinoma(HCC)and the adjacent normal tissues were analyzed with GEO2R,and the results and mir-10b target gene conducted the Venn diagram screening to obtain the intersecting target genes.Then the protein interactions were analyzed by using STRING and the core protein gene of miR-10b target gene was obtained.Results The miR-10b expression was significantly up-regulated in the HBV stably expressed HepG2.2.15 cells,and the difference was statistically significant(P<0.05);the target genes were mainly enriched in transcription regulation,transcription factor activity and related protein genes involving in transcription factor complex.miR-10b target genes were enriched in the proteoglycan,miRNA,and cAMP signaling pathways;the functions of the core proteins such as CREB1,NCOA6,NCOR2,PIK3CA,GATA3,MAPRE1 and TIAM1,and their functions involve the glucose homeostasis regulation,inflammation and occurrence and development of tumor.Conclusion HBV up-regulates the miR-10b expression,and the highly expressed miR-10b on the silencing of the target genes may play an important role in HBV infected liver diseases.
作者
梁俐兰
刘莉莉
覃超梅
梁斌
石明连
刘永明
苏何玲
LIANG Lilan;LIU Lili;QIN Chaomei;LIANG Bing;SHI Minglian;LIU Yongming;SU Heling(Teaching and Researching Section of Biochemistry and Molecular Biology,Guilin Medical University,Guilin,Guangxi 541100,China)
出处
《重庆医学》
CAS
2021年第21期3726-3731,共6页
Chongqing medicine
基金
国家自然科学基金项目(81560341、81460320)
广西壮族自治区自然科学基金项目(2018GXNSFAA138153)。
