■高通量测序用于筛选miRNA的表达差异,lncRNA,新诊断的免疫性血小板减少症(ITP)患者和健康对照者外周血CD19B中的mRNA和mRNA。本研究旨在探讨ceRNA网络在ITP患者CD19+B淋巴细胞功能失调发病机制中的调控作用。
■CD19+B淋巴细胞从ITP患者及其健康对应者的外周血样本中分离。高通量测序用于筛选miRNA的表达,lncRNA,和ITP患者和健康对照的mRNA,通过ceRNA网络分析。此外,qPCR用于验证miRNA的差异表达,lncRNA,和mRNA在ITP患者和健康对照中的表达。差异表达的miRNA,lncRNA,mRNA并对B淋巴细胞亚群进行了分析。
■对4例新诊断的ITP患者和4例健康对照者的CD19+B淋巴细胞进行测序和分析。有65个差异表达的lncRNA和149个mRNA形成ceRNA网络,表明12个lncRNA和136个差异表达的mRNA密切相关。同样,miR-144-3p,miR-374c-3p,miR-451a在ITP患者中高表达,如qPCR所证实,这与高通量序列结果一致。LOC102724852和CCL20在ITP患者中高表达,而LOC105378901、LOC112268311、ALAS2和TBC1D3F与健康对照相比,这与高通量序列结果一致。此外,miR-374c-3p的表达,LOC112268311、LOC105378901和CXCL3与B淋巴细胞亚群百分比相关。
■miRNA的ceRNA网络,lncRNA,外周血CD19+B淋巴细胞中的mRNA在ITP的发病机制中起着至关重要的作用。
High-throughput sequencing was used to screen expressing differences of miRNA, lncRNA, and mRNA in CD19+ B peripheral blood samples of newly diagnosed immune thrombocytopenia (ITP) patients and healthy controls. The study aimed to explore the regulatory role of ceRNA network in the pathogenesis of dysfunctional CD19 + B lymphocytes of ITP patients.
CD19+ B lymphocytes were isolated from peripheral blood samples of ITP patients and their healthy counterparts. High-throughput sequencing was used to screen for the expression of miRNA, lncRNA, and mRNA of ITP patients and healthy controls, which were analysed by the ceRNA network. Moreover, qPCR was used to verify the differential expression of miRNA, lncRNA, and mRNA in ITP patients and healthy controls. The correlation between differentially expressed miRNA, lncRNA, mRNA, and B lymphocyte subsets was also analysed.
The CD19+ B lymphocytes of 4 newly diagnosed ITP patients and 4 healthy controls were sequenced and analysed. There were 65 differentially expressed lncRNA and 149 mRNA forming a ceRNA network showed that 12 lncRNA and 136 differentially expressed mRNA were closely associated. Similarly, miR-144-3p, miR-374c-3p, and miR-451a were highly expressed in ITP patients, as confirmed by qPCR, which was consistent with the high-throughput sequence results. LOC102724852 and CCL20 were highly expressed in ITP patients, while LOC105378901, LOC112268311, ALAS2, and TBC1D3F were not as compared to healthy controls, which was consistent with the high-throughput sequence results. In addition, the expression of miR-374c-3p, LOC112268311, LOC105378901, and CXCL3 were correlated with the percentage of B lymphocyte subsets.
The ceRNA network of miRNA, lncRNA, and mRNA in peripheral CD19 + B lymphocytes plays an essential role in the pathogenesis of ITP.