基于Jurkat细胞分析circ_0006689对系统性红斑狼疮中IL2的作用及机制研究  

Effect and mechanism of circ_0006689 on IL2 in systemic lupus erythematosus based on Jurkat cells

作　　者：谭曦 韦悦 范润哥 李文宇[1] 侯伟 林有坤[1] Tan Xi;Wei Yue;Fan Runge;Li Wenyu;Hou Wei;Lin Youkun(Department of Dermatology and Venereal Diseases,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Key Laboratory of Thalassemia Prevention and Treatment,Chinese Academy of Medical Sciences,Nanning 530021,China;Guangxi Key Laboratory of Thalassemia Prevention and Treatment,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院皮肤性病科,南宁530021 [2]中国医学科学院地中海贫血防治研究重点实验室,南宁530021 [3]广西地中海贫血防治重点实验室,南宁530021

出　　处：《广西医科大学学报》2023年第6期953-959,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81760561,No.82260620)。

摘　　要：目的:在Jurkat细胞基础上初步探讨circ_0006689在系统性红斑狼疮(SLE)中调控白介素2(IL2)的作用机制。方法:使用慢病毒转染Jurkat细胞构建circ_0006689低表达株。用PMA、PHA活化低表达组(si-circ_0006689组)、病毒空载组(si-NC组)及未转染组(control组)细胞后通过实时荧光定量PCR(RT-qPCR)检测各组IL2、miR-95-5p的相对表达量;酶联免疫吸附试验(ELISA)法检测各组细胞IL2浓度;生物信息学分析预测circ_0006689的目标miRNA及IL2的目标miRNA;双荧光素酶报告实验检测circ_0006689与miR-95-5p、miR-95-5p与IL2的靶向关系。结果:Jurkat细胞活化后IL2分泌显著增加(P<0.05);下调circ_0006689后细胞IL2表达及分泌减少(P<0.05),miR-95-5p表达水平显著升高(P<0.05)。双荧光素酶报告实验提示circ_0006689与miR-95-5p、miR-95-5p与IL2存在结合作用。结论:在Jurkat细胞中低表达circ_0006689可抑制IL2的表达;miR-95-5p可分别与circ_0006689、IL2结合;circ_0006689可能在SLE中通过竞争性结合miR-95-5p影响IL2的表达及分泌。Objective:To preliminarily explore the effect and mechanism of circ_0006689 in regulating interleukin 2(IL2)in systemic lupus erythematosus(SLE)on the basis of Jurkat cells.Methods:Jurkat cells were transfected with lentivirus to construct circ_0006689 low expression strain;the relative expression levels of IL2 and miR-95-5p were determined by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)after PMA and PHA activated the cells in low expression group(si-circ_0006689 group),virus-free group(si-NC group)and untransfected group(control group).The concentration of secreted IL2 in each group was detected by enzyme-linked immunosorbent assay(ELISA);bioinformatics analysis predicted the target miRNA of circ_0006689 and the target miRNA of IL2;dual-luciferase reporter assay was used to detect the targeting relationship between circ_0006689 and miR-95-5p,and miR-95-5p and IL2.Results:IL2 secretion significantly increased after Jurkat cell activation(P<0.05);circ_0006689 was down-regulated,IL2 expression and secretion decreased(P<0.05),and the expression level of miR-95-5p increased significantly(P<0.05).The dual-luciferase reporter assay suggested the binding effect of circ_0006689 to miR-95-5p and miR-95-5p to IL2.Conclusion:Low expression of circ_0006689 in Jurkat cells can inhibit IL2 expression;miR-95-5p can bind to circ_0006689 and IL2,respectively;the circ_0006689 may affect IL2 expression and secretion by competitive binding to miR-95-5p in SLE.

关 键 词：系统性红斑狼疮 环状RNA 微小RNA 白介素2 

分 类 号：R593.24[医药卫生—内科学]