let-7c与其靶基因细胞因子在肺结核痰液中的表达及其功能分析  

Level of let-7c and target gene cytokine in sputum of patients with pulmonary tuberculosis and analysis of its function

作　　者：伊正君[1] 付玉荣[2] 吴晓燕[2] 许福亮[1] 

机构地区：[1]潍坊医学院附属医院检验科/临床检验诊断学省级重点实验室,潍坊261031 [2]潍坊医学院基础医学院病原生物学教研室,潍坊261053

出　　处：《中国人兽共患病学报》2013年第4期385-388,411,共5页Chinese Journal of Zoonoses

基　　金：国家自然科学基金项目(81100006);山东省自然科学基金(ZR2010HM073)联合资助~~

摘　　要：目的检测microRNA let-7c及其靶基因细胞因子在肺结核患者痰液中的含量变化,并用生物信息学技术和软件对let-7c的功能进行预测分析,为进一步阐明let-7c在结核感染中的作用奠定基础。方法用核酸杂交和定量PCR检测健康对照和活动性肺结核病人的痰液中let-7c的含量;用TargetScan与PicTar预测let-7c的靶基因,用DAVID数据库和Cyto-scape的BINGO插件对let-7c的预测靶基因进行GO注释分析和生物通路分析;用ELISA法检测let-7c靶基因细胞因子IL-10与IL-6在痰液中的水平变化。结果杂交和PCR结果表明:let-7c在活动性肺结核患者痰液中的含量均比健康对照组显著降低(P<0.05)。let-7c预测靶基因的GO注释结果富集在负性调节细胞间通讯等生物学过程及胞外基质构成等分子功能方面(P<0.05);在KEGG通路数据库中let-7c预测靶基因主要富集于MAPK、钙离子信号通路与TGF-beta信号通路等方面(P<0.05)。痰液中let-7c的靶基因细胞因子IL-10的含量显著增高。结论 let-7c在活动性肺结核感染病人痰液中的含量明显降低,let-7c靶基因细胞因子IL-10明显增高,let-7c的靶基因主要参与细胞信号通讯、刺激反应、信号转导等生物学过程。这些结果为进一步深入研究let-7c在结核感染中调控功能奠定了基础。Aim of the study was to analyze level of let-7c and target gene cytokine in sputum of patients with pulmonary tuberculosis, predict and analyze function of its target genes in order to lay foundation for further studying of its role in pulmo- nary tuberculosis. Sputum samples were collected from healthy controls and patients with active pulmonary tuberculosis. Level of let-7c was measured by nucleic acid hybridization and real-time reverse transcription-quantitative polymerase chain reaction （RT-qPCR）. Target genes of let-7c were predicted with TargetScan. GO annotation and pathway overrepresentation were fur- ther analyzed with DAVID database and BINGO of Cytoscape. ELISA was used to detect the level of target gene cytokines IL-10 and IL-6 in sputum. Result showed that level of let-7c obviously decreased in sputum of patients with active pulmonary tuberculosis compared with that in the healthy controls. The gene set mainly existed in negative regulation of cell communica- tion, response to endogenous Stimulus and negative regulation of signal transduction （GO biology process, P〈0. 01）, extracel- lular matrix structural constituent, growth factor binding, and transcription factor binding （GO molecular function, P〈0.05）. In KEGG pathway, the gene set mostly existed in MAPK signaling pathway, calcium signaling pathway, and TGF-beta signa- ling pathway （P〈0. 05）. Compared with the healthy controls, level of IL-10 obviously increased in sputum of patients with ac- tive pulmonary tuberculosis. Our results laid a foundation for further studies toward the role of let-7c in pulmonary tuberculosis infection.

关 键 词：肺结核 let-7c 靶基因 IL-10 生物信息学 

分 类 号：R378.91[医药卫生—病原生物学]