摘要
分析毒性结核分枝杆菌（Mycobacterium tuberculosis,M.tb）H37Rv刺激RAW264.7巨噬细胞的表达谱芯片以及筛选和鉴定M.tb H37Rv感染巨噬细胞RAW264.7后差异表达的lncRNA。首先,分析热灭活的H37Rv刺激RAW264.7细胞24h后lncRNA表达谱芯片,并对差异表达的lncRNA和mRNA进行生物信息分析,然后采用实时定量聚合酶链反应对16条在芯片中差异表达的lncRNA进行细胞水平验证;进一步在H37Rv感染小鼠的脾脏和肺脏中检测差异表达的lncRNA。结果显示,表达谱芯片中4 730条lncRNA的表达水平上调,9 558条lncRNA的表达水平下调。生物信息分析筛选的16条lncRNA,其染色体定位于附近蛋白质编码基因的基因间区或者与外显子区域有重叠,mRNA功能注释显示差异表达的mRNA主要集中于转录调节、磷酸化、凋亡等生化过程以及丝裂原活化蛋白激酶（MAPK）等抗结核的信号通路中。在H37Rv作用下的细胞水平和动物感染模型的组织中RT-q PCR验证出与芯片结果相同的4条lncRNA,其中上调的有3条,下调的有1条。研究中异常表达的lncRNA可为巨噬细胞在结核分枝杆菌感染中的功能紊乱提供线索,为后续的研究奠定基础,进一步的研究将集中于发掘lncRNA在宿主调控中发挥的功能。
To screen for differentially lncRNAs expression profile and predict their functional roles in the macrophage after Mycobacterium tuberculosis（ M. tb） stimulation. Firstly,microarray and bioinformatics analysis of lncRNA and mRNA expression profiles in RAW264. 7 macrophage after stimulation with M. tb for 24 h. Then,16 differentially expressed lncRNAs from microarray analysis were further verified by RT-q PCR using H37 Rv infected macrophages and mouse model. The results shown that the expression levels of 4 730 lncRNAs was upregulated,and the expression levels of 9 558 lncRNAs was down-regulated. 16 differentially expressed lncRNAs from microarray screening were associated with protein coding genes in adjacent locations. The mRNA function annotation analysis revealed that the mRNAs of differential expression were mainly concentrated in the biochemical process of transcriptional regulation,phosphorylation,apoptosis and MAPK signaling pathway which participating in the biochemical process of anti-tuberculosis. The expression trend of 4 lncRNAs in the i H37 Rv stimulated RAW264. 7 and mouse infection model was vertified as the same in both microarray and RT-q PCR analysis. Three of these 4 lncRNAs was up-regulated and one of them was down-regulated. The abnormal expression of lncRNAs may provide clues to the dysfunction of macrophages with M. tb infection,and further research will focus on the investigation of the function and regulation mechanism of lncRNA in M. tb infected macrophage.
作者
谭杨
刘胜
罗凤玲
章晓联
TAN Yang;LIU Sheng;LUO Feng-ling;ZHANG Xiao-lian(Department of Immunology, State Key Laboratory of Virology, Hubei Province Key Laboratory of Allergy and Immunology and Institute of Medical Research, Wuhan University School of Medicine,Wuhan 430071, China)
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2018年第5期1-9,共9页
China Biotechnology
基金
“十二五”国家重大传染病专项(2012ZX10003002-015)
“十三五”国家重大传染病专项(2017ZX10201301-006)资助项目
