桥本甲状腺炎患者血浆外泌体miRNA表达谱研究

Differential expression profiles of plasma exosomal miRNA in Hashimoto′s thyroiditis patients

目的检测桥本甲状腺炎患者(Hashimoto′s thyroiditis,HT)甲状腺功能正常组、HT合并亚临床甲状腺功能减退(SCH)组以及对照组外泌体微小RNAs(microRNAs,miRNAs),探讨HT病程中不同甲状腺状态下的miRNA差异表达谱及其相应的生物学功能。方法选取HT甲状腺功能正常者、HT合并SCH患者及健康志愿者各3人,提取外周血浆外泌体进行miRNA高通量测序,统计分析3组间差异表达miRNA,并进行靶基因预测、基因本体功能(GO)及京都基因与基因组百科全书通路(KEGG pathway)分析。结果得到75条差异表达miRNA(log2FC>1且Q-value≤0.001),数据挖掘后获得hsa-miR-324-3p、hsa-miR-370-3p、hsa-miR-423-5p、hsa-miR-532-3p等10条核心miRNA。GO分析表明,差异miRNA的靶基因主要参与细胞过程、生物调节、代谢过程等生物学过程,具备结合、催化活性、转录调节活性等分子功能。KEGG分析得到的通路主要为环磷酸腺苷(cAMP)信号通路、人类表皮生长因子受体(human epidermal growth factor receptor,ErbB)信号通路、缺氧诱导因子-1(hypoxia inducible factor-1,HIF-1)信号通路等。结论HT患者血浆外泌体miRNA较健康人存在差异表达,其在甲状腺功能正常与SCH两种不同状态中也存在差异表达。hsa-miR-324-3p、hsa-miR-370-3p、hsa-miR-423-5p及hsa-miR-532-3p等miRNA可能通过cAMP、ErbB及HIF-1等信号通路在HT病程进展中发挥一定的作用。

Objective To detect the exosomal miRNA in the cases of Hashimoto′s thyroiditis(HT)with euthyroidism,HT combined with subclinical hypothyroidism and healthy control,and to explore the differential expression profiles of miRNA in different thyroid states in the course of HT and their corresponding biological functions.Methods Three cases of HT with euthyroidism,3 cases of HT with subclinical hypothyroidism and 3 cases of healthy volunteers were enrolled in this study.The miRNA were extracted from peripheral plasma of these 9 samples,then high-throughout miRNA sequencing technology was performed for screening differential expression profiles,which followed by target gene prediction,Gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes pathway(KEGG pathway)analysis.Results A total of 75 differentially expressed miRNAs were found(log2FC>1 and Q-value≤0.001),and 10 core miRNAs were obtained through data mining,including hsa-miR-324-3p,hsa-miR-370-3p,hsa-miR-423-5p,hsa-miR-532-3p,etc.The results of GO enrichment analysis showed that these differential miRNA targeted genes mainly participated in a variety of biological processes such as cellular process,biological regulation and metabolic processes.Besides,they were involved in binding,catalytic activity,transcription regulation activity and other molecular functions.cAMP signaling pathway,ErbB signaling pathway and HIF-1 signaling pathway were the main pathways obtained by enrichment analysis of KEGG pathway.Conclusion There are differential expression profiles of exosomal miRNA in HT patients,and also in the two different states of normal thyroid function and subclinical hypothyroidism.MiRNAs such as hsa-miR-324-3p,hsa-miR-370-3p,hsa-miR-423-5p and hsa-miR-532-3p may play an important role in the progression of HT through cAMP signaling pathway,ErbB signaling pathwa and HIF-1 signaling pathway.