【摘要】 目的：检测急性髓系白血病（AML）患者外周血中外泌体miR-4286的表达情况，阐明miR-4286在AML诊断中的价值。方法：收集45例AML初诊患者（AML组）和35名同期健康体检者（对照组）外周血标本，利用超速离心法提取外周血中外泌体，离心产物分别采用透射电子显微镜（TEM）观察形态，纳米颗粒示踪分析（NTA）法检测粒子直径，Western blotting法检测表面标志物CD63和CD9蛋白的表达情况；采用实时荧光定量PCR (RT-qPCR)法检测2组受试者外泌体中miR-4286表达水平；利用受试者工作特征（ROC）曲线计算曲线下面积（AUC），分析miR-4286诊断AML的灵敏度和特异度。结果：TEM观察，提取的外周血外泌体为外观圆形、中间凹陷的膜状结构，直径为30～100 nm;NTA法，外泌体颗粒直径主要分布在30～150 nm;Western blotting法检测外泌体均表达CD63和CD9蛋白，具备外泌体的典型特征。RT-qPCR法，AML组患者外周血中外泌体miR-4286表达水平明显高于对照组（Z=-5.543, P<0.01）。ROC曲线分析，当外泌体中miR-4286表达水平以3.182为临界值时，AUC为0.862 9 (95%CI:0.774 7,0.937 1,P<0.01)，灵敏度为66.67%，特异度为97.14%，具有良好的诊断效能。结论：外周血外泌体中miR-4286对AML的诊断效能较高，可作为AML的诊断标志物，具有较高的临床诊断价值。更多还原

【Abstract】 Objective:To detect the expression of miR-4286 in exosome in peripheral blood of the patients with acute myeloid leukemia(AML), and to clarify the value of miR-4286 in the diagnosis of AML. Methods:The peripheral blood samples from 45 newly diagnosed patients with AML(AML group) and 35 concurrent healthy subjects(control group) were collected. The exosomes from serum of the subjects in two groups were obtained by over speend contrifugation; the morphology of the centrifuged products was observed under transmission electron microscope(TEM);and the particle diameter was detected by nanoparticle tracink analysis(NTA); the expressions of surface markers CD63 and CD9proteins were analyzed by Western blotting method;the expression levels of miR-4286 in exosome of the subjects in two groups were detected by real-time fluorescence quantitative PCR(RT-qPCR) method;the receiver operating characteristic(ROC) curve was used to calculate the area under curve(AUC); the sensitivity and specificity of miR-4286 in the diagnosis of AML were analyzed. Results:The TEM results showed that the extracted exosomes in peripheral blood showed a membrane structure with roundness in apperance and concave in the middle, and the diameter was about 30-100 nm;the NTA results showed that the particle sizes of exosomes were mainly distributed at 30-150 nm;the Western blotting results showed that the exosomes all expressed CD63 and CD9 proteins,with the typical characteristics of the exosomes. The RT-qPCR results showed that the expression level of exosome miR-4286 in peripheral blood of the patients in AML group was significantly higher than that in control group(Z=-5. 543,P<0. 01). The ROC curve analysis showed that when the critical value of expression level of miR-4286 in exosome was 3. 182, the AUC was 0. 862 9(95%CI:0. 774 7,0. 937 1, P<0. 01), the sensitivity was 66. 67%,the specificity was 97. 14%,and the diagnostic efficacy was good. Conclusion:The miR-4286 in exosome in peripheral blood has high diagnostic efficiency for AML;it can be used as a diagnostic marker for AML, and has high clinical diagnostic value.更多还原