3种环状RNA在系统性红斑狼疮中的表达及临床意义

Expression and clinical significance of 3 kinds of circular RNA in systemic lupus erythematosus

目的探讨三种环状RNA(circRNA)在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者中的表达及临床意义,为揭示SLE的临床诊断、治疗提供新的思路和方向。方法筛选35例SLE患者(实验组)及25例健康对照者(对照组),提取外周血单个核细胞(PBMCs),采用反转录定量PCR(RT-qPCR)检测circADCY9,circGARS,circMCTP2的表达水平,分析3种circRNA表达水平与临床活动度指标之间的相关性,采用受试者工作特征曲线(ROC)评价3种circRNA的诊断价值,通过核酸外切酶RNaseR处理检测3种circRNA分子的稳定性及耐受程度,选取circADCY9分子PCR验证其稳定性。结果(1)RT-qPCR分析结果显示circADCY9和circGARS在SLE患者PBMCs中表达上调,circMCTP2表达下调(P值均<0.0001);(2)SLE患者circADCY9,circGARS表达水平与SLE疾病活动指数(SLEDAI)评分呈正相关,与补体C3呈负相关,circMCTP2表达水平与SLEDAI评分呈负相关,与补体C3呈正相关;(3)SLE患者组和健康对照组circADCY9、circGARS、circMCTP2分子ROC曲线下面积(AUC)分别为0.8500、0.8267、0.8956;(4)RNaseR处理前后3种circRNA表达情况较为稳定,PCR显示:circRNA相较于mRNA对RNaseR的耐受性更好。结论SLE患者PBMCs中circADCY9,circGARS,circMCTP2表达明显异常,且稳定性好,与SLE疾病活动度具有相关性。

Objective To explore the expression and significance of 3 circular RNAs(circRNA),that is,circADCY9,circGARS and circMCTP2,in the patients with systemic lupus erythematosus(SLE),so as to provide new ideas and directions for its clinical diagnosis and treatment.Methods Peripheral blood mononuclear cells(PBMCs)were extracted from the SLE patients(n=35)and healthy volunteers(n=25,control group).The expression levels of above 3 circRNAs were detected by real-time quantitative PCR(RT-qPCR),and their correlations with SLE Disease Activity Index(SLEDAI)were analyzed.The diagnostic value of the 3 circRNAs was evaluated by receiver operating characteristic(ROC)curve.Their stability and tolerance were detected using ribonuclease R(RNaseR)treatment,and the stability of circADCY9 was further verified by PCR analysis.Results(1)The levels of circADCY9 and circGARS were up-regulated,while that of circMCTP2 was down-regulated(All P values were<0.0001)in the SLE patients than the healthy volunteers.(2)In SLE patients,the expression levels of circADCY9 and circGARS were positively correlated with the score of SLEDAI,and negatively with the level of complement C3.By contrast,the circMCTP2 level was negatively correlated with SLEDAI score and positively with complement C3 level.(3)The area under the curve(AUC)for circADCY9,circGARS and circMCTP2 were 0.8500,0.8267 and 0.8956,respectively.(4)The expression levels of the 3 circRNAs remained steady before and after RNaseR treatment,and the result of PCR showed that the tolerance of circADCY9 to RNaseR was better than that of ADCY9 mRNA.Conclusion The expression of circADCY9,circGARS and circMCTP2 is significantly abnormal in the PBMCs of SLE patients,and shows good stability,which is correlated with the activity of SLE.