摘要
目的:基于转录组测序技术分析原发性血小板增多症(ET)血小板活化相关基因,寻找与ET血栓形成相关的潜在靶点。方法:收集ET患者和健康人的血液标本进行转录组测序,选取差异表达的lncRNA、miRNA和mRNA,构建lncRNA-miRNA-mRNA调控网络。使用基因本体论(GO)和京都基因与基因组百科全书(KEGG)对调控网络中的差异mRNA进行富集分析。应用实时荧光定量PCR(Real-time PCR)方法验证关键信号通路上的差异mRNA。结果:共鉴定出差异表达的32个lncRNA(3个上调、29个下调)、16个miRNA(8个上调、8个下调)和35个mRNA(27个上调、8个下调)。其中5个lncRNA、12个miRNA和19个mRNA构成调控网络。KEGG富集分析显示,差异mRNA与血小板活化信号通路相关,与血小板活化相关的差异mRNA有6个,分别为F2R、ITGA2B、ITGB1、ITGB3、PTGS1和GP1BB,均表达上调。RT-PCR验证结果显示,与健康人相比,ET患者5个mRNA包括F2R、ITGA2B、ITGB1、ITGB3和GP1BB表达水平上调,与转录组测序结果一致,而PTGS1表达无显著差异。结论:ET患者差异mRNA与血小板活化通路相关,F2R、ITGA2B、ITGB1、ITGB3和GP1BB mRNA可作为ET血小板活化相关的新靶点。
Objective:To analyze the genes related to platelet activation in essential thrombocythemia(ET)based on transcriptome sequencing technology(RNA-seq),and to explore the potential targets related to ET thrombosis.Methods:Blood samples from ET patients and healthy individuals were collected for RNA-seq,and differentially expressed lncRNAs,miRNAs,and mRNAs were selected to construct a lncRNA-miRNA-mRNA regulatory network.Differential mRNAs in the regulatory network were enriched and analyzed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).The real-time PCR method was applied to validate differential mRNAs on crucial signaling pathways.Results:A total of 32 lncRNAs(3 up-regulated,29 down-regulated),16 miRNAs(8 up-regulated,8 down-regulated),and 35 mRNAs(27 up-regulated,8 down-regulated)were identified as differentially expressed.Among them,5 lncRNAs,12 miRNAs,and 19 mRNAs constituted the regulatory network.KEGG enrichment analysis showed that the differential mRNAs were related to the platelet activation signaling pathway,and there were 6 differential mRNAs related to platelet activation,namely F2R,ITGA2B,ITGB1,ITGB3,PTGS1,and GP1BB,which were all up-regulated in their expression.RT-PCR results showed that the expression of five mRNAs including F2R,ITGA2B,ITGB1,ITGB3,and GP1BB were upregulated in ET patients compared with healthy subjects,and consistent with RNA-seq results,while PTGS1 expression was not significantly different.Conclusion:Differential mRNAs in ET patientsarerelated to the platelet activation pathway,andF2R,ITGA2B,ITGB1,ITGB3,andGP1BB mRNA smayserve as novel targets associated with platelet activation in ET.
作者
孙妍
杨二鹏
李雨蒙
牛继聪
赵霈
刘为易
陈卓
王明镜
范腾
胡晓梅
SUN Yan;YANG Er-Peng;LI Yu-Meng;NIU Ji-Cong;ZHAO Pei;LIU Wei-Yi;CHEN Zhuo;WANG Ming-Jing;FAN Teng;HU Xiao-Mei(Graduate School of Beijing University of Chinese Medicine,Beijing 100029,China;Department of Hematology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;VIP Inpatient Department,Sun Yat-sen University Cancer Center,Guangzhou 510060,Guangdong Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2024年第6期1814-1821,共8页
Journal of Experimental Hematology
基金
国家自然科学基金面上项目(82174360)
中国中医科学院科技创新工程重大攻关项目(CI2021A01702,CI2021A01707)。
