目的 探索结核性胸膜炎(tuberculous pleurisy,TP)患者胸腔积液差异表达微小核糖核酸(microRNA,miRNA)及其与自噬的关系.方法 选取作者医院就诊的10例TP患者(病例组)和10例其他胸腔积液患者(对照组),进行胸水总RNA提取、miRNA测序分析,以检测miRNAs表达谱的差异表达;进一步选取同期就诊的20例TP患者和20例非TP患者的胸腔积液验证miRNAs表达谱.通过逆转录聚合酶链反应(reverse transcription polymerase chain re-action,RT-PCR),受试者工作特征(receiver operating characteristic,ROC)曲线分析,真实性评价证实了其中6个表达不同的miRNAs的诊断意义.选取miR-21-5p预测其自噬相关基因之间、自噬蛋白之间的互作关系.结果 芯片系统分析筛选出差异表达miRNAs 73个.RT-PCR结果显示,与对照组比较,miR-3148、miR-4701-3p在病例组中上调;miR-223-3p、miR-23a-3p、miR-7a-5p、miR-21-5p 在病例组中下调.ROC 曲线结果显示,miR-7a-5p、miR-23a-3p、miR-21-5p、miR-223-3p、miR-3148、miR-4701-3p对 TP 的诊断性能曲线下面积(area under the curve,AUC)分别为 0.787、0.944、0.832、0.878、0.919、0.732.通过生物信息学分析同时结合人类自噬调节数据库(Human Autophagy Moderator Data-base,HAMdb)进行预测,显示与miR-21-5p相关度最高的前5位基因分别是BECN1、MAPK1、FOXO3、HSPA8和RAB11A.结论 结核性胸腔积液(tuberculous pleural effusion,TPE)中的miRNAs对TP有一定诊断价值,其中6个miRNAs为诊断治疗TP提供了新的标志物和潜在的治疗靶点,并且推测miR-21-5p可能通过自噬相关基因来调节自噬,影响TP的发病过程和结局,值得进一步研究.
Identification of microRNA Expression Profile and Diagnostic Markers in Tuberculous Pleurisy
Objective To explore the differentially expressed microRNA(miRNA)in pleural effusion of patients with tuberculous pleurisy(TP)and its relationship with autophagy.Methods A total of 10 patients with TP(case group)and 10 other patients with pleural effusion(control group)attending the authors'hospital were selected for total RNA extraction from pleural fluid and miRNA sequencing analysis in order to detect the differential expression of miR-NAs expression profiles;pleural effusion from 20 patients with TP and 20 non-TP patients attending the same period of time were further selected for verification of miRNAs expression profiles.Subsequently,the diagnostic significance of 6 of the differently expressed miRNAs was confirmed by reverse transcription polymerase chain reaction(RT-PCR),receiver operating characteristic(ROC)curves and authenticity assessment of another portion of the population sample.Finally,the study selected miR-21-5p to predict its autophagy-related genes and autophagy proteins interactions.Results A total of 73 differentially expressed miRNAs were screened by chip system analysis.RT-PCR results showed that compared with control group,miR-3148,miR-4701-3p in case group up-regulated;miR-223-3p,miR-23a-3p,miR-7a-5p and miR-21-5p were down-regulated in the case group.The ROC curve results showed that the area under the curve of the diagnos-tic performance of miR-7a-5p,miR-23a-3p,miR-21-5p,miR-223-3p,miR-3148,miR-4701-3p for TP were 0.787,0.944,0.832,0.878,0.919,0.732 respectively.The top 5 genes with the highest correlation with miR-21-5p as predic-ted by bioinformatics analysis in conjunction with Human Autophagy Moderator Database(HAMdb)were BECN1,MAPK1,FOXO3,HSPA8 and RAB11 A.Conclusion The miRNAs in tuberculous pleural effusion(TPE)have diagnos-tic value for TP,6 miRNAs in the study,provides new markers and potential therapeutic targets for the diagnosis and treatment of TP,and it is hypothesized that miR-21-5p may regulate autophagy through autophagy-related genes affecting the pathogenesis and outcome of TP,which de-serves further study.
NETL
NSTL
万方数据
