Hsa_circ_0000880在阿尔兹海默病患者血浆中的表达及诊断价值

Expression and diagnostic value of hsa_circ_0000880 in plasma of patients with Alzheimer′s disease

目的:探索hsa_circ_0000880作为阿尔茨海默病(AD)诊断生物标志物的价值。方法:采集28例AD患者(AD组)以及28例正常人(对照组)的血浆,采用qRT-PCR法检测hsa_circ_0000880和miR-214-3p的表达水平。采用ROC曲线评价hsa_circ_0000880诊断AD的临床价值。分析AD患者的简易智力状态检查量表(MMSE)、蒙特利尔认知评估(MoCA)评分与血浆hsa_circ_0000880水平的相关关系。采用双荧光素酶报告实验验证hsa_circ_0000880与miR-214-3p之间的关系。SH-SY5Y细胞分为si-Circ和si-NC组,分别转染hsa_circ_0000880 siRNA和对照siRNA,采用qRT-PCR法检测miR-214-3p的表达水平。结果:与对照组比较,AD组血浆hsa_circ_0000880水平升高(P=0.015);其对AD诊断ROC曲线的AUC为0.886,95%CI为0.773~0.999。AD患者血浆hsa_circ_0000880水平与MMSE、MoCA评分呈负相关(r=-0.646、-0.601,P<0.001)。AD患者血浆miR-214-3p与hsa_circ_0000880水平呈负相关(r=-0.053,P=0.002)。双荧光素酶报告实验结果显示,miR-214-3p mimic和Wt-hsa_circ_0000880共转染的细胞荧光素酶活性降低。抑制hsa_circ_0000880表达后SH-SY5Y细胞中miR-214-3p的表达升高(P=0.048)。结论:Hsa_circ_0000880可能通过影响miR-214-3p的表达,从而在AD中发挥病理作用;hsa_circ_0000880可作为AD诊断的生物标志物。

Aim:To explore whether hsa_circ_0000880 can be used as a potential biomarker for AD diagnosis.Methods:Plasma was collected from 28 AD patients(AD group)and 28 normal subjects(control group),and the expression level of hsa_circ_0000880 was detected by qRT-PCR.The clinical value of hsa_circ_0000880 for the diagnosis of AD was evaluated using the ROC curve.The correlation between MMSE and MoCA scores and hsa_circ_0000880 expression in AD patients was analyzed.Dual luciferase reporter assay was used to verify the relationship between hsa_circ_0000880 and miR-214-3p.SH-SY5Y cells were divided into si-Circ and si-NC groups and transfected with hsa_circ_0000880 siRNA and control siRNA,respectively.The expression level of miRNA-214-3p was detected by qRT-PCR.Results:The expression of hsa_circ_0000880 in plasma was significantly higher in the AD group than that in the control group(P=0.015).The AUC value of ROC curve for AD diagnosis was 0.886,95%CI was 0.773-0.999.Hsa_circ_0000880 expression level was negatively correlated with MMSE and MoCA scores in AD patients(r=-0.646,-0.601,P<0.001).Relationship between plasma hsa_circ_0000880 and miR-214-3p in patients with AD was negatively correlated(r=-0.053,P=0.002).The results of dual luciferase reporter gene assay showed that the luciferase activity of miR-214-3p mimic and Wt-hsa_circ_0000880 co-transfected cells decreased.After inhibiting the expression of hsa_circ_0000880,the expression of miR-214-3p in SH-SY5Y cells increased(P=0.048).Conclusion:Hsa_circ_0000880 is abnormally up-regulated in plasma of AD patients and may affect the expression of miR-214-3p,thus exerting a pathological role in AD,suggesting that hsa_circ_0000880 may serve as a biomarker for AD diagnosis.