环状RNA在口腔白斑和口腔扁平苔藓中的共同表达  

Co-expression of circRNA in oral leukoplakia and oral lichen planus

作　　者：杨靖雯 宋雨翰 徐思明 葛姝云[1] 周海文[1] YANG Jingwen;SONG Yuhan;XU siming;GE Shuyun;ZHOU Haiwen(Department of Oral Mucosal Diseases,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,College of Stomatology,Shanghai Jiao Tong University National center for Stomatoloy,National Clinical Research Center for oral diseases,Shanghai Key Laboratory of Stomatology,Shanghai Research Institute of Stomatology,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院口腔黏膜病科,上海交通大学口腔医学院,国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海200011

出　　处：《口腔疾病防治》2023年第11期774-785,共12页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：国家自然科学基金项目(81170968);上海交通大学医学院第九人民医院与上海交通大学口腔医学院研究型学科基金项目资助(KQYJXK2020)。

摘　　要：目的 探讨环状RNA在口腔白斑(oral leukoplakia,OLK)和口腔扁平苔藓(oral lichen planus,OLP)疾病发生发展中的共同表达,为研究OLK与OLP的发病机制及癌变机制提供基础。方法 本研究获得医院伦理审批。采用高通量测序技术检测OLK、OLP、口腔鳞状细胞癌和正常口腔黏膜组织中差异表达的circRNA,采用qRTPCR、酶耐受实验和sanger测序筛选并验证OLK与OLP中共同显著差异表达circRNA,并对关键circRNA扩大样本量验证;采用GO和KEGG富集分析差异circRNA可能涉及的生物学过程及信号通路;采用TargetScan与MiRanda预测circRNA下游的miRNA并绘制ceRNA网络。结果 OLK与OLP共同差异表达的circRNA共49个,包括30个上调和19个下调的circRNA;经验证,筛选的3个上调circRNA(circHLA-C,circTTN,circRNF13)和2个下调circRNA(circSEPN2,circALDH3A2)表达趋势与测序结果一致。其中circHLA-C是具有反式剪接位点的关键circRNA,经扩大样本量验证,circHLA-C在OLK与OLP中显著上调。ROC曲线分析显示,OLK中circHLA-C曲线下面积为0.955,OLP中circHLA-C曲线下面积为0.988。GO功能富集显示与免疫功能生物学过程密切相关,KEGG通路分析中富集分数最高的信号通路是“自然杀伤细胞介导的细胞毒性”,HLA-C在这些通路中显著富集,ceRNA网络分析表明circHLA-C可能与多种癌症相关miRNA相互作用,如hsa-miR-26a-5p、hsa-miR-129-5p和hsa-miR-29a-3p。结论 OLK与OLP存在相同差异表达的circRNA,circHLA-C是OLK与OLP共有的最显著上调的circRNA;circHLA-C可能会成为OLP与OLK诊断和预后的潜在生物标志物。Objective To find any differentially expressed circRNAs in oral leukoplakia(OLK)and oral lichen planus(OLP),to investigate the possible role of circRNAs in the pathogenesis of these two diseases.Methods This study obtained hospital ethical approval.High-throughput sequencing was used to detect differentially expressed circRNAs in OLK,OLP,oral squamous cell carcinoma and normal oral mucosal tissues.CircRNAs were verified by qRT-PCR,enzyme tolerance assays and Sanger sequencing.GO functional analysis and KEGG pathway analysis were performed to predict the functions of circRNAs in OLP.TargetScan and miRanda were applied to predict targeted miRNAs and mRNAs of circRNAs,and ceRNA networks were mapped.Results A total of 49 circRNAs were differentially expressed in OLK and OLP together,including 30 upregulated and 19 downregulated circRNAs.The five circRNAs confirmed with RT-qPCR,including circHLA-C,circRNF13,circTTN,circSEPN2 and circALDH3A2,were all abnormally expressed in OLK and OLP,among which circHLA-C was a key circRNA with trans splice sites,which was validated by expanding the sample size.ROC curve analysis showed that the area under the circHLA-C curve for predicting OLK was 0.955,and the area under the circHLA-C curve for predicting OLP was 0.988.GO functional analysis showed enrichment of many biological processes related to the immune process.The KEGG pathway with the highest enrichment score was"Natural killer cell mediated cytotoxicity".HLA-C was significantly enriched in these processes/pathways.CeRNA network analysis showed that circHLA-C interacted with a variety of miRNAs,such as hsa-miR-26a-5p,hsa-miR-129-5p,and hsa-miR-29a-3p.Conclusion Many circRNAs were differentially expressed in both OLK and OLP,circHLA-C being the most elevated.CircHLA-C is valuable for the early diagnosis of OLK and OLP and may serve as a potential biomarker for the diagnosis and prognosis of OLK and OLP.

关 键 词：环状RNA circHLA-C 口腔白斑病 口腔扁平苔藓 口腔黏膜潜在恶性疾病 癌变 生物标志物 高通量测序 

分 类 号：R78[医药卫生—口腔医学]