微小RNA-29a在肺结核患者血清中的表达及其生物学功能预测分析  

Expression of miR-29a in serum of patients with pulmonary tuberculosis and prediction of itsfunction with bioinformatics analysis

作　　者：付玉荣[1] 伊正君 李建花 高昆山 

机构地区：[1]潍坊医学院基础医学院病原生物学教研室,山东261053 [2]检验系临床检验诊断学省级重点实验室附属医院检验科

出　　处：《中华结核和呼吸杂志》2013年第3期186-190,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金（81100006）;山东省自然科学基金（ZR2010HM073）

摘　　要：目的分析微小RNA-29a（miR-29a）在肺结核患者血清中的表达，并对miR-29a的靶基因进行功能预测分析，为深入研究miR-29a与结核病感染调控机制的关系及其生物学功能奠定基础。方法病例组为未经治疗的活动性肺结核患者65例，对照组为健康志愿者45例，清晨空腹抽取静脉血，分离血清。用TRIzol试剂获取血清中总RNA，对RNA样本进一步纯化，通过紫外分光光度计和凝胶电泳检测RNA提取质量。用核酸杂交和荧光定量PCR检测miR-29a在血清中的表达。运用TargetScan与PicTar软件综合预测miR-29a的靶基因，取两者的交集作为靶基因。采用DAVID数据库对miR-29a预测的靶基因集合进行基因本体论（GO）注释描述分类，并分别提取GO的3类注释信息。采用网络调控分析软件Cytoscape中的插件BINGO进行GO注释的显著性分析。对靶基因集合进行生物通路富集分析。结果核酸杂交结果表明，病例组血清中miR-29a表达（854±93）显著高于对照组（80±22），差异有统计学意义（t=3．541，P〈0．05）。荧光定量PCR结果表明，病例组血清中miR-29a表达（1．35±0．62）显著高于对照组（0．18±0．07），差异有统计学意义（t=2．987，P〈0．05）。预测miR-29a的靶基因集合分别富集在转录调控等生物学过程、金属离子结合活性等分子功能及细胞外基质等细胞组分；miR-29a的靶基因集合显著富集于KEGG通路数据库中的黏附、细胞外基质受体相互作用等7个信号转导通路和Biocarta通路数据库中的整联蛋白信号通路、钙黏蛋白信号通路和Wnt信号通路等信号通路中。结论活动性肺结核患者血清中miR-29a表达显著上调，miR-29a的靶基因参与细胞黏附、转录调控等生物学过程。Objective To analyze the expression of miR-29a in serum of patients with pulmonary tuberculosis, and to predict and analyze function of its target genes for further studying of its biological function and regulatory mechanism in tuberculosis （TB）. Methods Fasting venous blood samples were collected from 65 untreated patients with active pulmonary TB （ case group） and 45 healthy controls （ control group） in the morning, respectively, and then sera were isolated. Total RNA was extracted with TRIzol reagent and was further purified. RNA quality was measured by gel electrophoresis and ultraviolet spectrophotometer. Nucleic acid hybridization and real-time reverse transcription-quantitative polymerase chain reaction （RT-qPCR） were performed to investigate expression of miR-29a in serum. Both TargetSean and PieTar software were used to predict comprehensively target genes of miR-29a and their intersection was regarded as target genes of miR-29a. Gene ontology of target genes was analyzed with DAVID database and three category annotations were extracted, respectively. GO overrepresentation was further analyzed by BINGO of Cytoscape. Enriched pathways of target genes were analyzed. Results The hybridization result showed that miR-29a was increased in serum of the ease group （854±93） compared to the control group （80±22） （t = 3. 541, P 〈0. 05）. The PCR result showed that compared to the control group （0. 18 ±0. 07）, miR-29a was increased in serum of the case group （ 1.35 ± 0. 62 ） （ t = 2. 987, P 〈 0. 05 ）. Thetarget genes were mainly involved in biological processes including regulation of-transcription, molecular function including metal ion binding, and cellular components including extracellular matrix, respectively. In the KEGG pathway, the target gene set was significantly enriched in the 7 signaling pathways including adhesion, ECM-receptor interaction and so on. In the PANTHER pathway, the gene set mostly existed in integrin signaling pathway, cadherin signaling path

关 键 词：结核 肺 微RNAS 靶基因修复 计算生物学 

分 类 号：R5[医药卫生—内科学]