新型循环提取方法研究miRNA-21在NSCLC中的预测价值  

The predictive value of circulating miRNA-21 in NSCLC by constructing a novel circulating miRNA extraction method

作　　者：荣昊[1] 任伟宏[2] 李延卿[2] 何鑫 冯倩[2] 李文博[2] 冯慧洁[2] RONG Hao;REN Weihong;LI Yanqing;HE Xin;FENG Qian;LI Wenbo;FENG Huijie(Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450000,China;Department of Clinical Laboratory,First Affiliated Hospital,Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450000,China)

机构地区：[1]河南中医药大学,郑州450000 [2]河南中医药大学第一附属医院检验科,郑州450000

出　　处：《检验医学与临床》2018年第22期3367-3370,3376,共5页Laboratory Medicine and Clinic

摘　　要：目的构建新型血清miRNA提取方法,并评估血清外泌体miRNA-21表达水平与非小细胞肺癌(NSCLC)患者病理与临床特征的相关性。方法收集30例NSCLC患者(试验组)和10例健康志愿者(对照组)。从每例受试者外周血中离心出两份等量的外周血血清,设为A、B两组,每组为相同的40例血清标本。在miRNA提取方法方面,A组为直接从血清中提取miRNA-21,B组先富集血清外泌体,再从外泌体中提取miRNA-21,然后采用实时荧光定量PCR法分别检测A、B两组血清miRNA-21表达水平,通过分析两组血清中各自试验组、对照组的检测结果是否存在明显差异,来比较两种血清miRNA提取方法的优劣;并进一步分析血清外泌体miRNA-21表达水平与NSCLC患者病理分型、病理分期、TNM分期的相关性。结果 NSCLC患者血清外泌体中miRNA-21的表达水平较健康人明显上调(P<0.05),上调倍数为2.93倍,受试者工作曲线显示其预测NSCLC风险价值高,曲线下面积为0.867(P<0.05),95%CI为0.721～1.000;而通过直接从血清中提取miRNA-21的方法,NSCLC患者和健康人表达水平差异无统计学意义(P>0.05)。NSCLC患者外泌体miRNA-21表达水平与病理分期、TNM分期相关(P<0.05),与病理分型无关(P>0.05)。结论血清外泌体miRNA-21表达水平具有更高的预测NSCLC风险的价值,并且与NSCLC病理分期、TNM分期相关。Objective Take the expression of miRNA-21 in peripheral blood of patients with non-small cell lung cancer（NSCLC） is significantly increased as the entry point to establish a method of miRNA extraction that serum exosomes are enriched first and miRNA in exosomes are then extracted;And evaluate the correlation between the level of miRNA-21 in serum exosomes and the pathological and clinical features of NSCLC patients. Methods Thirty NSCLC patients（experimental group） and 10 healthy volunteers（control group） were enrolled.Peripheral blood serum was collected from the subjects,and the serum of each subject was divided evenly into two groups.In group A,miRNA-21 was directly extracted from serum;In group B,serum exosomes are enriched first and miRNA-21 in exosomes are then extracted.And then using real-time fluorescence PCR to detect the expression of serum miRNA-21 in group A and group B.By comparing the results of experimental group with the control group in two groups of serum,to compare the quality of the two methods of serum miRNA extraction;and then analyze the relevance of serum exosome miRNA-21 and NSCLC pathological type,pathological stage,TNM stage. Results The expression level of miRNA-21 in serum exosomes of NSCLC patients was significantly higher than that of healthy people（ P 〈0.05）,and the up-regulation was 2.93 times.The working curve of the subjects showed high value in predicting the risk of NSCLC,area under the curve was 0.867.The 95% confidence interval （ CI ） ranged from 0.721 to 1.000.While there was no significant difference between NSCLC patients and healthy controls（ P 〉0.05） by directly extracting miRNA-21 from serum.The expression of miRNA-21 in exosomes of NSCLC was correlated with pathological stage（ P 〈0.05）,TNM stage（ P 〈0.05）,but not with pathological type（ P 〉0.05）. Conclusion Compared with serum miRNAs,serum exosome miRNAs have higher detection efficiency.Serum exosome miRNA-21 expression level can be used as a biomarker that predicts t

关 键 词：NSCLC 血清 外泌体 MIRNA 

分 类 号：R446.11[医药卫生—诊断学]