Graves病患者外周血单个核细胞中环状RNA表达谱分析

Expression profile of peripheral blood mononuclear cells derived circular RNA in patients with Graves′disease

目的:探讨Graves病患者外周血单个核细胞(PBMCs)中环状RNA(circRNA)的表达谱及其潜在的诊断价值。方法:选取5例Graves病患者和5例健康对照者,采用二代高通量测序技术分析PBMCs中circRNA表达谱。另选取40例Graves病患者和40例健康对照者验证差异表达的circRNA;根据CircInteractome数据库预测可能与差异表达的circRNA相结合的微小RNA(miRNA);运用生物信息学分析差异表达的circRNA潜在的生物学功能;ROC曲线分析差异表达的circRNA对Graves病的诊断价值。结果:二代高通量测序数据显示,与健康对照相比,Graves病患者PBMCs中circRNA有23215个表达上调,13695个表达下调,其中显著差异表达(差异倍数≥2,P≤0.05)的有669个,包括360个上调和309个下调的circRNA。实时荧光定量PCR验证显示hsa_circ_0114427、hsa_circ_0007470、hsa_circ_0048232、hsa_circ_0002672表达水平与高通量测序结果一致,且两组间差异具有统计学意义(P<0.01和P<0.05),hsa_circ_0039161和hsa_circ_0002600表达两组间无明显差异。GO富集分析显示,上调的circRNA与高分子代谢、细胞内膜结合细胞器、转移酶激活等过程关系密切。KEGG分析显示,差异表达的circRNA可能参与甲状腺激素信号、磷脂酰肌醇信号系统、非小细胞肺癌等相关信号通路。ROC曲线显示hsa_circ_0114427具有较大的曲线下面积。结论:Graves病患者PBMCs中存在明确差异表达的circRNA,hsa_circ_0114427可能与Graves病的发生发展有关。

Objective:To explore the expression profile of circular RNA(circRNA)in peripheral blood mononuclear cells(PBMCs)of patients with Graves′disease and its potential diagnostic value.Methods:Five patients with Graves′disease and five healthy controls were selected to analyze the expression profile of circRNA in PBMCs by next generation high-throughput sequencing.Another 40 patients with Graves′disease and 40 healthy controls were selected to verify the differential expression of circRNA;MicroRNAs(miRNAs)that may bind to differentially expressed circRNA were predicted according to CircInteract database;Bioinformatics was used to analyze the potential biological functions of differentially expressed circRNA;ROC curve analysis of the differential expression of circRNA in Graves′disease.Results:Next generation high-throughput sequencing data showed that,compared with healthy controls,there were 23215 up-regulated and 13695 down-regulated circRNA in patients with Graves′disease,of which 669 were significantly differentially expressed,including 360 up-regulated and 309 down regulated circRNA(difference multiple≥2,P≤0.05).Hsa_circ_0114427,hsa_circ_0007470,hsa_circ_0048232 and hsa_circ_0002672 were confirmed by real-time PCR,and the expression level was consistent with that of high-throughput sequencing,and there were statistical significance between the two groups(P<0.01 and P<0.05),but there was no difference in expression of hsa_circ_0039161 and hsa_circ_0002600.GO enrichment analysis showed that the up-regulated circRNA was closely related to polymer metabolism,cell membrane binding organelles,and transferase activation.KEGG analysis showed that the differentially expressed circRNA might be involved in thyroid hormone signaling,phosphatidylinositol signaling system,non-small cell lung cancer and other related signaling pathways.ROC curve showed hsa_circ_0114427 has the biggest area under the curve.Conclusion:There are clearly differentially expressed circular RNAs in PBMCs of patients with Graves′disease.Hsa_circ_0114427 may be related to the occurrence and development of Graves′disease.