RAW264.7细胞自噬相关circRNA检测及分析  

Detection and analysis of autophagy-related circRNA in RAW264.7 cells

作　　者：张爱君 李艳宁 高倩 屈昱良[1] 蒋丹[1] 王丽燕 徐广贤[1,3] ZHANG Aijun;LI Yanning;GAO Qian;QU Yuliang;JIANG Dan;WANG Liyan;XU Guangxian(School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,China;Department of Laboratory Medicine,Affiliated Hospital of Air Force Military Medical University,Xi'an 710000,China;Department of Medical Laboratory Medicine,School of Medical Technology,Guangdong Medical University,Dongguan 523000,China)

机构地区：[1]宁夏医科大学临床医学院,银川750001 [2]空军军医大学附属医院检验科,西安710000 [3]广东医科大学医学技术学院医学检验系,东莞523000

出　　处：《免疫学杂志》2022年第2期177-184,共8页Immunological Journal

基　　金：宁夏重点研发计划(社发领域)重点项目。

摘　　要：目的探讨环状RNA(circRNA)表达谱在RAW264.7细胞自噬模型中的差异表达及其在细胞自噬过程中的调控作用。方法利用雷帕霉素(rapamycin,RAPA)和3-甲基腺嘌呤(3-methyladenine,3-MA)分别作用于小鼠单核巨噬细胞RAW264.7细胞,建立自噬诱导模型和自噬抑制模型;提取各组细胞总RNA,基于高通量测序技术获得差异表达的circRNA;运用生物信息学分析对circRNA可能结合的微小RNA(miRNA)及其可能参与自噬的分子通路进行预测。结果在RAPA组和3-MA组中呈现相反表达趋势的circRNA共有42个,差异显著的前5个circRNA分别是circRNAmmu_circ_0000353、mmu_circ_0001704、mmu_circ_0013108、mmu_circ_0001528和mmu_circ_0015842,分别对其结合匹配值较高的3个miRNA位点进行预测,GO分析表明这些miRNA与转录调控、炎症反应及免疫应答相关;KEGG分析表明与多条自噬相关通路有关。结论本研究筛选到的circRNA以及与之结合的miRNA与细胞自噬的发生发展相关,可以此为靶点进行自噬相关的研究,并为自噬调控机制提供基础。To investigate the differential expression of circular RNA(circRNA)expression profile in the autophagy model of RAW264.7 cells and its regulatory role in the process of autophagy,rapamycin(RAPA)and 3-methyladenine(3-MA)were used to treat RAW264.7 cells for establishing autophagy induction model and autophagy inhibition model.The total RNA of each group was extracted,and differentially expressed circRNA was obtained based on high-throughput sequencing technology.Bioinformatics analysis was performed to predict the microRNA(miRNA)binding to circRNA and the molecular pathways participating in autophagy.A total of 42 circRNAs showed opposite expression trends in the RAPA group and 3-MA group.The top 5 circRNAs with significant differences were mmu_circ_0000353,mmu_circ_0001704,mmu_circ_0013108,mmu_circ_0001528 and mmu_circ_0015842,while 3 miRNA sites with higher binding matching values were predicted.GO analysis showed that these miRNAs are related to transcriptional regulation,inflammation and immune response;KEGG analysis showed that they are related to multiple autophagy-related pathways.Taken together,the circRNAs screened in this study and the miRNA bound to them are related to the occurrence and development of autophagy,which can be used as a target for autophagy-related research and provide a basis for research of autophagy regulation mechanism.

关 键 词：细胞自噬 circRNA 高通量测序 生物信息学 

分 类 号：R392.12[医药卫生—免疫学]