3-O-C_(12)-HSL作用下Mo-DCs的circRNA表达谱与功能富集分析  

The circRNA expression profile and functional enrichment of Mo-DCs under the effect of 3-O-C_(12)-HSL

作　　者：陈敏怡 陈蕾妃 李有强 张云燕 CHEN Minyi;CHEN Leifei;LI Youqiang;ZHANG Yunyan(Department of Pediatric Dentistry,School and Hospital of Stomatology,Guangdong Engineering Research Center of Oral Restoration and Reconstruction&Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine,Guangzhou Medical University,Guangzhou,Guangdong 510182,China;Hexian Memorial Affiliated Hospital of Southern Medical University,Guangzhou,Guangdong 511400,China;Department of Dentistry,Guangzhou Women and Children Medical Center,Guangzhou,Guangdong 510627,China)

机构地区：[1]广州医科大学口腔医学院·附属口腔医院儿童口腔科·广东省口腔组织修复与重建工程技术研究中心·广州市口腔再生医学基础与应用研究重点实验室,广东广州510182 [2]南方医科大学附属何贤纪念医院检验科,广东广州511400 [3]广州市妇女儿童医疗中心口腔科,广东广州510627

出　　处：《热带医学杂志》2024年第10期1361-1366,1378,F0002,共8页Journal of Tropical Medicine

基　　金：国家自然科学基金(81801973);广东省自然科学基金(2022A1515012255);广东省医学科研基金(A2023400,A2020560);广州市基础研究计划市校(院)联合资助基础与应用基础研究项目(202201020629);广州市番禺区科技计划项目(2021-Z04-003)。

摘　　要：目的探讨单核细胞来源树突状细胞(Mo‐DCs)在N-3-氧化十二烷酰-L-同型丝氨酸内酯(3‐O‐C_(12)‐HSL)作用下其环状RNA(circRNA)表达谱的影响及功能富集。方法取健康志愿者新鲜外周血,经Fi‐coll密度梯度离心及免疫磁珠分选获得CD14+单核细胞,重组人白细胞介素-4(hIL‐4)、重组人粒-巨噬细胞集落刺激因子(hGM‐CSF)细胞因子诱导分化为Mo‐DCs,将细胞用0.1μg/mL脂多糖(LPS)(LPS组、阳性对照)、0.1%二甲基亚砜(DMSO)(DMSO组、阴性对照)或25μmol/L 3‐O‐C_(12)‐HSL和0.1μg/mL LPS(3‐O‐C_(12)‐HSL组)分别处理。采用Arraystar CircRNA芯片筛选三组差异表达的circRNA,并对差异表达circRNA靶基因作功能富集分析。结果circRNA微阵列分析检测到12967个circRNA在Mo‐DCs中表达,根据P<0.05和FC≥1.2的标准进行筛选,与LPS组相比,3‐O‐C_(12)‐HSL组有122个circRNA显著上调,77个显著下调;与DMSO组相比,3‐O‐C_(12)‐HSL组有64个circRNA显著上调,122个显著下调。GO和KEGG富集分析表明,差异表达的circRNA亲本基因主要富集于转化生长因子-β、趋化因子、ErbB等信号通路上,主要参与树突状细胞分化、Fcγ受体介导的吞噬作用及淋巴细胞分化调节等过程。通过ENCORI数据库预测大量miRNA与circRNA存在结合位点,并构建了ceRNA网络图。结论3‐O‐C_(12)‐HSL影响了Mo‐DCs成熟过程中circRNA的表达。Objective To investigate the effect of N-3-dodecanoyl-L-homoserine lactone(3-O-C_(12)-HSL)on the expression profile of circular RNA(circRNA)and its functional enrichment in monocyte-derived dendritic cells(Mo-DCs).Methods Fresh peripheral blood from healthy volunteers was collected,and CD14+monocytes were obtained through Fi-coll density gradient centrifugation and immunomagnetic bead sorting.The cells were differentiated into Mo-DCs induced by human interleukin 4(hIL-4)and human granulocyte monocyte colony stimulating factor(hGM-CSF)cytokines.The cells were treated with 0.1μg/mL lipopolysaccharide(LPS)(LPS group,positive control),0.1%dimethyl sulfoxide(DMSO)(DMSO group,negative control),or 25μmol/L 3-O-C_(12)-HSL and 0.1μg/mL LPS(3-O-C_(12)-HSL group)separately.Arraystar circRNA chip was used to screen three groups of differentially expressed circRNAs,and functional enrichment analysis was performed on differentially expressed circRNA target genes.Results circRNA microarray analysis detected 12967 circRNAs expressed in Mo-DCs.Screening was based on the criteria of P<0.05 and FC≥1.2.Compared with the LPS group,122 circRNAs in the 3-O-C_(12)-HSL group were significantly up-regulated and 77 were significantly down-regulated;compared with the DMSO group,64 circRNAs were significantly up-regulated and 122 were significantly down-regulated in the 3-O-C_(12)-HSL group.GO and KEGG enrichment analysis showed that differentially expressed circRNA parental genes were mainly enriched in signaling pathways such as transforming growth factor-β,chemokines,and ErbB,and were mainly involved in dendritic cell differentiation and Fcγreceptor-mediated processes such as phagocytosis and lymphocyte differentiation regulation.A large number of binding sites for miRNA and circRNA were predicted through the ENCORI database,and a ceRNA network diagram was constructed.Conclusion 3-O-C_(12)-HSL could affect the expression of circRNA during the maturation of Mo-DCs.

关 键 词：环状RNA circRNA芯片测序 生物信息学分析 N-3-氧化十二烷酰-L-同型丝氨酸内酯 树突状细胞 

分 类 号：R318.04[医药卫生—生物医学工程]