外周血ADAM19 DNA甲基化生物标记物对慢性阻塞性肺疾病的诊断价值  

Diagnostic Value of Peripheral Blood ADAM19 DNA Methylation Biomarkers in Chronic Obstructive Pulmonary Disease

作　　者：王霞 侯嘉[2] WANG Xia;HOU Jia(College of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,China;Department of Respiratory and Critical Care Medicine,the General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学临床医学院,银川750004 [2]宁夏医科大学总医院呼吸与危重症医学科,银川750004

出　　处：《宁夏医科大学学报》2022年第8期823-828,共6页Journal of Ningxia Medical University

基　　金：宁夏回族自治区重点研发项目(2021BEG03079);宁夏自然科学基金项目(2020AA03401)。

摘　　要：目的筛选与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)发病相关的ADAM19 DNA甲基化生物标记物。方法分析Illumina 850K芯片全基因组甲基化测序结果(8例COPD患者和8例健康者),筛选差异甲基化位点cg07575896及靶基因ADAM19。收集2020年10月至2021年4月在宁夏医科大学总医院呼吸与危重症医学科住院的102例COPD患者(COPD组)和本院健康体检中心的61例健康者(对照组)的外周全血;Agena质谱法验证外周血DNA甲基化;RT-PCR检测外周血ADAM19 mRNA相对表达;ROC曲线分析评估ADAM19作为COPD诊断效能;分析ADAM19基因表达与COPD患者肺功能分级、呼吸衰竭类型的关系。结果利用Illumina 850K芯片随机抽取两组中各8例研究对象的外周血进行甲基化测序,结果显示,与对照组相比,COPD组甲基化位点cg07575896发生高甲基化(Δβ=0.08,P<0.05),Agena质谱法分析结果显示,与对照组相比,COPD组甲基化位点cg07575896中CpG4发生高甲基化(P<0.05);利用RT-PCR检测结果示,与对照组比较,COPD组ADAM19 mRNA表达上调(P<0.001);利用ROC曲线分析评估外周血ADAM19基因对COPD的预测价值,曲线下面积(AUC)为0.78(95%CI:0.71~0.85,P<0.001);COPD患者GOLD分级肺功能越低,ADAM19的mRNA表达量越高(P均<0.05);COPD患者呼吸衰竭程度越重ADAM19的mRNA表达量越高(P<0.05)。结论外周血ADAM19可能成为COPD早期诊断和病情评价的生物标记物。Objective To investigate the epigenetics of the gene ADAM19 associated with chronic obstructive pulmonary disease(COPD)as a peripheral blood methylation biomarker in COPD.Methods Illumina 850K microarray whole genome methylation sequencing results(8 COPD cases and 8 normal subjects)were analyzed to screen for differential methylation sites cg07575896 and target gene ADAM19.102 COPD patients(COPD group)hospitalized in the Department of Respiratory and Critical Care Medicine at the General Hospital of Ningxia Medical University from October 2020 to April 2021 and our health peripheral whole blood from 61 healthy people(control group)in the health screening center.Agena mass spectrometry was used to verify DNA methylation in peripheral blood.RT-PCR was used to detect the relative expression of ADAM19 mRNA in peripheral blood.ROC curve analysis was used to assess the efficacy of ADAM19 as a diagnosis of COPD,and analysis of the relationship between ADAM19 gene expression and lung function classification and respiratory failure type was analyzed in COPD patients.Results The results of peripheral blood of 8 subjects methylation sequencing using Illumina 850K chip in two randomized selected groups showed that methylation site cg07575896 was hypermethylated in the COPD group compared to the control group(Δβ=0.08,P<0.05),and the results of Agena mass spectrometry analysis showed that CpG4 was hypermethylated in the methylation site cg07575896 in the COPD group compared to the control group(P<0.05).The results using RT-PCR showed that ADAM19 mRNA expression was upregulated in the COPD group compared with the control group(P<0.001),the predictive value of the ADAM19 gene in peripheral blood for COPD was assessed using ROC curve analysis,and the area under the curve(AUC)was 0.78(95%CI:0.71-0.85,P<0.001),the lower the lung function of COPD patients with GOLD classification,the higher the mRNA expression of ADAM19(P all<0.05);the more severe the degree of respiratory failure of COPD patients,the higher the mRNA expression of

关 键 词：慢性阻塞性肺疾病 ADAM19 DNA甲基化 生物标记物 

分 类 号：R563.3[医药卫生—呼吸系统]