新疆维吾尔族T2DM患者外周血circRNA差异表达初步研究

A preliminary study on the differential expression of circRNA in peripheral blood of Uygur patients with T2DM in Xinjiang

目的探讨外周血中环状RNAs(circRNAs)在新疆维吾尔族2型糖尿病(T2DM)分子生物学发病机制的意义。方法采用高通量核苷酸测序,从维吾尔族正常对照者(NC组)和维吾尔族T2DM患者各5例(T2DM组)的外周血单核细胞中筛选出差异表达的circRNAs;扩大样本量(NC组20例、T2DM组22例)利用实时荧光定量PCR(RT-qPCR)进一步验证差异表达的circRNAs。结果高通量核苷酸测序共筛选出134个差异表达的circRNAs,其中87个上调,47个下调;挑选表达明显上调hsa-circ-0139634、has-circ-0005414及明显下调hsa-circ-0032491、hsa-circ-0002922。通过RT-qPCR在两组维吾尔族人群外周血中进一步验证,hsa-circ-0139634、hsa-circ-0032491及hsa-circ-0002922在外周血样本中的差异表达有统计学意义,与测序结果一致;hsa-circ-0139634的ROC曲线下面积是0.926[95%CI(0.839~1.000),P<0.001],约登指数为0.833,敏感性为1.000,特异性为0.833,截断值为0.169;has-circ-0005414的ROC曲线下面积是0.575[95%CI(0.370~0.779),P=0.910],约登指数为0.278,敏感性为0.412,特异性为0.867,截断值为5.749;hsa-circ-0032491的ROC曲线下面积是0.316[95%CI(0.086~0.440),P=0.017],约登指数为0.056,敏感性为1.000,特异性为0.056,截断值为0.230;hsa-circ-0002922的ROC曲线下面积是0.263[95%CI(0.078~0.428),P=0.014],约登指数为0.080,敏感性为0.955,特异性为0.125,截断值为0.011。结论维吾尔族T2DM患者与NC人群存在差异表达的circRNAs谱;hsa-circ-0139634在维吾尔族T2DM分子生物学发病机制方面有潜在价值。

Objective To explore the significance of circular RNAs(circRNAs)underlining the molecular biological pathogenesis of Uyghur type 2 diabetes(T2DM)by observing the difference of circRNAs expression in the peripheral blood of Uyghur T2DM patients.Methods High-throughput nucleotide sequencing was utilized to screen out differentially expressed circRNAs from peripheral blood mononuclear cells of 5 Uyghurs volunteers with healthy adults and 5 Uyghurs with T2DM.To further verify the differentially expressed circRNAs,additional samples from 20 NC Uyghur volunteers and 22 T2DM Uyghur patients were analyzed by real-time fluorescent quantitative-PCR(RT-qPCR).Results A total of 134 differentially expressed circRNAs were screened out by high-throughput nucleotide sequencing,of which 87 were up-regulated and 47 were down-regulated.hsa-circ-0139634 and has-circ-0005414 were obviously up-regulated and hsa-circ-0032491 and hsa-circ-0002922 were obviously down-regulated and they were selected for further verification by RT-qPCR of the additional samples.The study found that the differential expression of hsa-circ-0139634、hsa-circ-0032491、hsa-circ-0002922 in peripheral blood samples was statistically significant,which was consistent with the sequencing results.Among them,the area under the receiver operating characteristic(ROC)curve of hsa-circ-0139634 was 0.935[95%confidence interval(0.884-1.000),P<0.001],Youden index was 0.867,sensitivity was 1.000,specificity was 0.867 and cutoff value was 0.169;the area under the receiver operating characteristic(ROC)curve of has-circ-0005414 was 0.575[95%confidence interval(0.370-0.779),P=0.910],Youden index was 0.278,sensitivity was 0.412,specificity was 0.867 and cutoff value was 5.749;the area under the ROC curve of hsa-circ-0032491 was 0.316[95%confidence interval(0.086-0.440),P=0.017],Youden index was 0.056,sensitivity was 1.000,specificity was 0.056 and cutoff value was 0.230;the area under the ROC curve of hsa-circ-0002922 was 0.263[95%confidence interval(0.078-0.428),P=0.014],Youden index was 0.080,sensitivity was 0.955,specificity was 0.125 and cutoff value was 0.011.Conclusion Uyghur T2DM patients and NC populations have differentially expressed circRNAs profiles.Of note,hsa-circ-0139634 has potential value in the molecular biological pathogenesis of Uyghur T2DM.