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目的了解结核病患者血清miRNA差异表达谱,并分析其功能,探索miRNA参与结核病易感性的作用机制。方法选取新疆喀什市维吾尔族活动性肺结核患者30例(病例组)和维吾尔族健康人28例(对照组),用Agilent表达谱芯片对病例组和对照组中的各9例血清miRNA进行检测,筛选差异表达的miRNA,并用qRTPCR进行验证,用生物信息学方法预测差异miRNA的靶基因,并对靶基因进行基因本体论(GO)分析和通路富集分析。结果芯片筛选出86个差异miRNA,其中43个表达上调,43个表达下调。对其中2个差异miRNA(miR-223-3p和miR-23a-3p)进行qRT-PCR验证,结果表明2个miRNA均在病例组中表达下调(P<0.05)。生物信息学软件预测到交集靶基因1 200个,GO分析表明预测的靶基因主要富集在蛋白质结合、蛋白磷酸化、转录调控、细胞周期调控及信号转导等生物学过程。通路分析表明靶基因主要参与到PI3K-AKT信号通路、癌症中的转录误调节、NF-κB等信号通路调节。结论新疆维吾尔族结核病患者血清中存在差异表达的miRNA,差异miRNA通过其靶基因参与多个生物学过程和信号通路调节,与结核病感染和宿主免疫调控有关。
Abstract：Objective To understand the differential miRNAs expressed in the sera of patients with tuberculosis in Xinjiang Uyghur and to explore their function,and the role in the susceptibility of tuberculosis.Methods 30 Uyghur patients with active tuberculosis(case group)and 28 Uyghur healthy controls(control group)were selected from Kashgar,and their serum were obtained,9specimens from case group and9 from control group were used to detect and filtrate the differential serum miRNAs by Agilent expression profile chip,and the remainder specimens of all the participants were used to verify the selected miRNAs using quantitative real time polymerase chain reaction(qRT-PCR).Target genes of differential miRNAs were predicted using bioinformatics methods,and gene ontology(GO)analysis and pathway enrichment analysis were performed by online tools.Results 86 differential miRNAs were screened out,of which 43 miRNAs were up-regulated and other 43 miRNAs were down-regulated.Two miRNAs of them,miR-223-3p and miR-23a-3p were verified by qRT-PCR method,and relative expression of them were shown to be all lower in case group than that in control group(P <0.01).1200 target genes of intersection of the two miRNAs were predicted by bioinformatics software,and gene ontology analysis indicated that these target genes were mainly enriched in protein binding,phosphoprotein,transcription regulation,cell cycle regulation and signal transduction.Pathway analysis showed the target genes participated in signal path regulation such as PI3K-AKT,transcriptional dysregulation in cancer and NF-κB pathway.Conclusion There were differential miRNAs in serum of patients with tuberculosis in Xinjiang Uyghur,which took part in many biological processes and signal pathways through their target genes,and thus could play the role of regulation in mycobacterium tuberculosis infection and host immunity.
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基本信息:
中图分类号:R440;R52
引用信息:
[1]孟存仁,葛金莲,王家路,等.新疆维吾尔族结核病患者血清差异表达miRNA筛选及功能分析[J].新疆医科大学学报,2017,40(07):971-976.
基金信息:
国家自然科学基金(81460323);; 新疆重大疾病医学重点实验室开放课题(SKLIB-XJMDR-2014-6)
2017-03-09
2017
2017-04-06
2017
1
2017-07-15
2017-07-15