口腔扁平苔藓组织环状RNA表达谱的筛选与初步探讨

Identification of circular RNA expression profiles in oral lichen planus

目的 :构建人口腔扁平苔藓(oral lichen planus,OLP)组织的环状RNA(circular RNA,circRNA)表达谱,筛选并验证口腔扁平苔藓组织中异常表达的circRNA,为疾病的诊断和治疗提供理论基础。方法:选择2018年9月—12月就诊于上海交通大学医学院附属第九人民医院口腔黏膜病科的口腔扁平苔藓初诊患者6例,同期健康志愿者6例,利用高通量测序技术对口腔扁平苔藓和正常口腔黏膜组织中RNA进行测序和鉴定分析,分析组间circRNA的表达差异,然后利用qRT-PCR对分析结果进行验证。采用SPSS 24.0软件包对数据进行统计学分析,最后应用生物信息学技术GO(Gene Ontology)富集分析和KEGG(Kyoto Encyclopedia of Genes and Genomes)通路分析差异基因的功能及相关通路。结果:测序结果显示,与健康口腔黏膜组织相比,在口腔扁平苔藓组织中共检测出85种差异表达2倍以上的circRNA,其中66种高表达,19种低表达。分别取上调、下调最显著的3个circRNA,在更多样本中进行qRTPCR验证,其结果与测序结果一致。结论:口腔扁平苔藓组织与正常口腔黏膜组织之间存在差异表达的circRNAs,这些circRNAs可能参与口腔扁平苔藓的致病调控,是潜在的疾病新型诊断和治疗的生物学标志物。

PURPOSE:To construct the expression profile of circular RNA(circRNA) in human oral lichen planus(OLP),and to identify and validate the differentially expressed circRNA in oral lichen planus tissues and provide theoretical basis for the diagnosis and treatment of this disease.METHODS:Six patients newly diagnosed with OLP from September to December 2018 in the Department of Oral Mucosal Diseases,Shanghai Ninth People’s Hospital and 6 healthy volunteers were enrolled in this study.RNA sequencing and evaluation in OLP tissues and normal oral mucosa were performed by high-throughput RNA sequencing technology,and the differences between groups were analyzed.qRT-PCR was used to validate the results.Statistical analysis was conducted with SPSS 24.0 software package.Finally,bioinformatics techniques GO(Gene Ontology) enrichment analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes) pathway were used to analyze the functions and related pathways of the dysregulated genes.RESULTS:According to the sequencing results,85differentially expressed circRNAs with fold change > 2 were identified in OLP tissues compared to the normal oral mu-cosa,including 66 upregulated circRNAs and 19 downregulated circRNAs.Three circRNAs with the most significant upregulation and down-regulation were selected for qRT-PCR verification in expanded samples,and the results were consis-tent with the sequencing results.Bioinformatics analysis suggested that the differentially expressed circRNAs may play an important role in the occurrence and progression of oral lichen planus.CONCLUSIONS:Differentially expressed circRNAs between oral lichen planus tissues and normal oral mucosa were identified,which may be involved in the pathogenic mechanism of oral lichen planus and could be potential biomarkers for diagnosis and treatment of this disease.