外周血CD14＋单核细胞微RNA在结核分枝杆菌潜伏感染和肺结核中的差异表达  

Differential expression of microRNA profiles in peripheral blood CD14＋ monocytes between latent tuberculosis infection and pulmonary tuberculosis patients

作　　者：林巧[1] 钟红剑[1] 邹容容[2] 刘腊香[1] 郑军[1] 张国良[2] 陈心春[2] 周伯平[2] 唐瑛[1] 

机构地区：[1]深圳市宝安区慢性病防治院综合门诊,518126 [2] 深圳市第三人民医院肝病研究所

出　　处：《中华临床感染病杂志》2015年第1期20-25,共6页Chinese Journal of Clinical Infectious Diseases

基　　金：广东省医学科研基金项目（B2013352）;深圳市战略性新型产业发展专项资金项目（ZDSY20120614101029732）

摘　　要：目的 分析结核分枝杆菌潜伏感染（LTBI）和肺结核患者外周血CD14＋单核细胞微RNA（miRNA）表达谱的差异.方法 选择深圳市宝安区慢性病防治院和深圳市第三人民医院2013年6月至2014年2月收治的肺结核患者31例,同时选择上述医院体检科的LTBI者31例.从肺结核和LTBI人群中各挑选6例（男、女各3例）进行miRNA芯片检测,筛选出差异表达的miRNA.选取具有代表性的差异表达的miRNA,采用TaqMan qPCR检测验证其在两组人群（各25例）中表达的差异.应用受试者工作特征（ROC）曲线评价差异表达的miRNA诊断肺结核的价值.应用miRFocus靶基因预测集成数据库对差异表达的miRNA进行靶基因预测,并对预测的靶基因进行GO和KEGG通路分析.结果 与LTBI人群相比,肺结核患者CD14＋单核细胞中表达上调2倍以上的miRNA有4个,表达下调2倍以上的miRNA有36个.对差异表达的miRNA进行聚类分析发现,差异表达的40个miRNA分子可分为2个基因簇.从2个基因簇中各选取一个具有代表性的miRNA分子（miR-378和miR-483-5p）进行TaqMan qPCR验证.结果显示,miR-378在肺结核患者中的表达水平为4.17 ±0.25,显著高于其在LTBI人群中的表达水平（2.31 ±0.24,=5.25,P＜0.01）;miR-483-5p在肺结核患者中的表达水平为1.71±0.16,显著低于其在LTBI人群中的表达水平（2.97 ±0.15,t =5.45,P＜0.01）.ROC曲线分析显示,miR-378诊断肺结核的敏感性为0.76,特异性为0.72;miR-483-5p诊断肺结核的敏感性为0.84,特异性为0.76.生物信息学分析显示miR-378和miR-483-5p的靶基因主要参与细胞增殖、凋亡、抗原提呈、信号转导等过程.结论 LTBI和肺结核人群外周血CD14＋单核细胞miRNA表达谱存在显著差异,其中miR-378和miR-483-5p可以作为有效分子标识鉴别诊断两组人群.Objective To screen differentially expressed microRNAs （miRNAs） in peripheral blood CD14 ＋ monocytes between latent tuberculosis infection （LTBI） and pulmonary tuberculosis patients.Methods Thirty-one patients with pulmonary tuberculosis and 31 patients with LTBI were enrolled from Shenzhen Bao＆#39; an Chronic Diseases Prevent and Treatment Hospital and Shenzhen Third People＆#39; s Hospital during June 2013 and February 2014.Differentially expressed miRNAs were detected by using a miRNA chip in 6 pulmonary tuberculosis patients and 6 LTBI patients （male 3,female 3）,and TaqMan qPCR test was performed to verify the differentially expressed miRNAs in two groups （25 for each）.Receiver operating characteristic （ROC） curve was used to evaluate the differentially expressed miRNAs in diagnosis of pulmonary tuberculosis.Target genes of differentially expressed miRNAs were forecasted using miRFocus database,and GO term and KEGG pathway annotation were performed.Results There were 40 differentially expressed miRNAs in CD14 ＋ monocytes between LTBI and pulmonary tuberculosis patients,among which 4 had 〉 2-fold up-regulation and 36 had 〉 2-fold down-regulation in pulmonary tuberculosis patients.All differentially expressed miRNAs could be divided into two clusters.TaqMan qPCR test showed that the expression of miR-378 （up-regulated miRNA） in pulmonary tuberculosis patients was 4.17 ± 0.25,which was significantly higher than that in LTBI patients （2.31 ± 0.24,t =5.25,P 〈 0.01） ; the expression of miR-483-5p （down-regulated miRNA） in pulmonary tuberculosis patients was 1.7l ± 0.16,which was significantly lower than that in LTBI patients （2.97 ± 0.15,t =5.45,P 〈 0.01）.ROC curve analysis showed that the sensitivities and specificities of miR-378 and miR-483-5p in the diagnosis of pulmonary tuberculosis were 0.76,0.72 and 0.84,0.76,respectively.Bioinformatic analysis showed that the target genes of miR-378 and miR-483-5p mainly involved in cell proliferation,apoptosis,antigen

关 键 词：分枝杆菌 结核 结核 肺 微RNAS 微RNA-378 MicroRNA-378 

分 类 号：R521[医药卫生—内科学]