系统性红斑狼疮患者外周血单个核细胞中环状RNA has_circ_0044235的表达及诊断价值研究

Identification of circular RNAs hsa_circ_0044235 as novel biomarkers for systemic lupus erythematosus

目的检测系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中环状RNA has_circ_0044235的表达水平,探讨其在SLE诊断中的应用价值。方法采用实时荧光定量PCR法在30例SLE与20例HC中对has_circ_0044235进行验证,应用受试者工作特征曲线(ROC)曲线分析has_circ_0044235诊断SLE的敏感度和特异度,对45例SLE,30例RA及38例HC外周血PBMC has_circ_0044235表达水平及诊断价值进行双重验证测试,并用TargetScan和miRanda软件预测与has_circ_0044235相互作用的miRNA。结果对hsa_circ_0044235进行qRT-PCR验证发现,与HC相比,SLE患者PBMC中hsa_circ_0044235的平均表达水平明显下降(P<0.0001)与芯片结果一致;ROC曲线分析表明,SLE患者和HC的hsa_circ_0044235 ROC曲线下面积为0.873(95%CI:0.778-0.967;P<0.001),敏感性为70.00%,特异性为100.00%;扩大样本量(45例SLE患者、38例HCS和30例RA)验证hsa_circ_0044235的表达水平及对SLE的独立诊断效能发现,与HC和疾病对照组RA相比,SLE患者PBMC中hsa_circ_0044235平均表达水平明显下降(P<0.001);hsa_circ_0044235 ROC曲线下面积SLE患者和HCs比较为0.861;SLE患者和对照者(HC+RA)为0.845;SLE患者和疾病对照者RA为0.825;与hsa_circ_0044235相互作用的miRNA为hsa-miR-892a,hsa-miR-135b-5和hsa-miR-135a-5p,qRT-PCR检测这些miRNA发现SLE患者PBMC的hsa-miR-892a水平显著升高(P=0.0002),而hsa-miR-135b-5和hsa-miR-135a-5p水平无明显异常(P>0.0500)。结论circRNA的失调可能在SLE发病机制中起作用,且hsa_circ_0044235可能通过竞争吸附miR-875-3p影响靶基因的表达,进而影响SLE疾病的发生和发展,PBMC中hsa_circ_0044235水平可作为SLE诊断的潜在生物标志物。

Objective To detect the expression level of circular RNA has_circ_0044235 in peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythematosus(SLE)and to explore its value in the diagnosis of SLE.Methods Real-time fluorescent quantitative PCR was used to verify has_circ_0044235 expression in 30 patients with SLE and 20 patients with HC.ROC curve was analysised the sensitivity and specificity of circ_0044235 for diagnosis SLE,and double-validation test of 45 cases of SLE,30 cases of RA and 38HC peripheral blood PBMC was used to test_circ_0044235 expression level and diagnostic value,and TargetScan and miRanda software was used to predict has_circ_0044235 interacting miRNA.Results The average expression level of hsa_circ_0044235 in PBMC of SLE patients was significantly decreased(P<0.0001)compared with HC in qRT-PCR method.ROC curve analysis showed that the area under the ROC curve of hsa_circ_0044235 in SLE patients was 0.873(95%CI:0.778-0.967;P<0.0001),with a sensitivity of 70.00%and a specificity of 100.00%.Compared with HC and disease control Compared with RA,the average expression level of hsa_circ_0044235 in PBMC of SLE patients was significantly decreased(P<0.001)the area under the ROC curve of hsa_circ_0044235 was 0.861 for SLE patients and HCs,and 0.845 for SLE patients and controls(HC+RA)and SLE patients and RA was 0.825.The miRNAs interacting with hsa_circ_0044235 were hsa-miR-892a,hsa-miR-135b-5 and hsa-miR-135a-5p,qRT-PCR detection of these miRNAs revealed that the level of hsa-miR-892a increased significantly in the PBMC of SLE patients(P=0.0002),while the levels of hsa-miR-135b-5 and hsa-miR-135a-5p were not statistical difference(P>0.0500).Conclusion Dysregulation of circRNA may play a role in the pathogenesis of SLE,and hsa_circ_0044235 may affect the expression of target genes through competitive adsorption of miR-875-3p,which in turn affects the occurrence and development of SLE disease.The level of hsa_circ_0044235 in PBMC can be used as a potential biomarker for the diagnosis of SLE.