摘要
目的：筛选免疫性血小板减少症（ ITP）患者血浆差异表达微小RNA（ miRNA）。方法收集2012年6月至2013年9月天津市第一中心医院血液科就诊的25例ITP患者及同期20名健康体检者血浆，采用miRNA芯片检测血浆中miRNA表达谱，筛选ITP血浆差异表达的miRNA。通过靶基因预测程序TargetScan和miRanda进行潜在的靶基因预测，并进行基因本体和信号通路注释分析，探索可能与ITP发病相关的基因和信号通路，并用实时（ real-time） PCR对差异表达的miRNA进行验证。结果根据基因芯片结果，发现ITP患者血浆中存在29个差异表达的miRNA，其中上调15个，下调14个，通过靶基因预测程序TargetScan和miRanda预测出608个潜在的靶基因，基因本体分析显示参与生物学过程的基因占78.12％（475／608）；参与分子功能的基因占80.76％（491／608）；参与细胞组分的基因占87.66％（533／608），富集分析显示P＜0.05的基因本体有9个。通路分析显示157个信号通路与608个靶基因相关，富集分析P＜0.05的信号通路有25个。其中值得特别关注的信号通路是钙离子信号通路和 T 细胞受体信号通路。 real-time PCR 显示 ITP 患者血浆中miRNA4778-5p、miRNA4800-5p 显著高于健康对照，而 miRNA4707-5p、miRNA4721、miRNA3620-3p、miRNA378i显著低于健康对照（均P＜0.05），与芯片结果一致。结论 miRNA芯片初步筛选出ITP患者血浆miRNA差异表达谱，生物信息学分析提示钙离子信号通路和T细胞受体信号通路可能与ITP患者发病和进展有关。
Objective To screen the plasma microRNA （ miRNA ） profile of immune thrombocytopenia （ ITP ） patients.Methods Agilent 19.0 miRNA microarray was used to detect the expression profile of miRNA in plasma from 25 ITP patients and 20 healthy controls from June 2012 to September 2013.The software programs of TargetScan and miRanda were used for predicting target genes associated with differential miRNA.Then gene ontology （ GO ） and pathway analysis were performed to explore the genes and pathways involved in the pathogenesis of ITP.And differential miRNA was validated by real-time quantitative polymerase chain reaction （ PCR）.Results A genome-wide miRNA array revealed 29 differential miRNAs in the plasma samples of ITP patients including 15 up-regulated and 14 down-regulated miRNA.A total of 608 potential genes were predicted by TargetScan and miRanda.GO result showed that there were 475 （ 78.12%） , 491 （ 80.76%） and 533 （ 87.66%） genes respectively involved in biological process, molecular function and cellular component.Enrichment test showed 9 GO terms had significant difference （ P 〈0.05 ） .Pathway analysis showed that 157 pathways were associated with 608 genes.Enrichment test showed 25 pathways had significant difference （ P〈0.05 ）.As revealed by real-time PCR, the expressions of miRNA4778-5p and miRNA4800-5p became obviously up-regulated while those of miRNA4707-5p, miRNA4721, miRNA3620-3p and miRNA378i decreased （ all P 〈0.05 ）.The results agreed with those of microarray.Conclusions The plasma differential miRNA profiles are identified in ITP patients.And miRNA is involved in calcium signaling pathway and T cell receptor signaling pathway may be associated with ITP pathogenesis.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2014年第14期1083-1086,共4页
National Medical Journal of China
基金
天津市卫生局科技攻关项目（13KG106）
