食管鳞癌患者血浆与肿瘤组织DNA甲基化检测的应用价值  

作　　者：王长春[1] 毛伟敏[1] 凌志强[1] 

机构地区：[1]浙江省肿瘤医院浙江省肿瘤研究所,杭州310022

出　　处：《中华检验医学杂志》2012年第1期37-41,共5页Chinese Journal of Laboratory Medicine

基　　金：基金项目：浙江省科技计划资助项目（2009C33143）;浙江省自然科学基金资助项目（Y2080749,Y2091110）;人事部优秀留学回国人员科技活动项目资助课题（2008A004）;教育部优秀留学回国人员科研基金资助课题（2010609）

摘　　要：目的 研究食管鳞癌患者肿瘤组织和血浆多基因甲基化状态及其对食管鳞癌诊断的应用价值.方法 用甲基化特异性聚合酶链反应（methylation specific polymerase chain reaction,MSP）检测76例食管鳞癌患者肿瘤组织、配对癌旁正常组织、患者术前1d、术后7d血浆游离DNA中腺瘤性息肉瘤基因（adenomatous polyposis coli,APC）、维甲酸受体β2基因（retinoic acid receptor-beta2,RARβ2）、上皮细胞钙黏蛋白基因（E-cadherin,CDH1）、细胞周期蛋白依赖激酶抑制剂4A家族p16基因（cyclin-dependent kinase inhibitor4A,p16INK4α）、Ras相关家族蛋白1A基因（ras association domain family member 1A,RASSF1 A）甲基化,选取同期60名年龄、性别匹配的健康志愿者血浆DNA作对照.用x2检验分析各类组织与血浆标本中5个基因DNA甲基化率的差异；血浆与食管鳞癌组织标本DNA甲基化的相关性用Spearman相关分析；绘制受试者操作特征（receive operating characteristic,ROC）曲线评价单基因与5个基因联合检测诊断食管鳞癌的敏感度、特异度.结果 食管鳞癌组肿瘤组织中的APC、RARβ2、CDH1、p16INK4α、RASSF1A基因甲基化阳性率分别为44.7％ （34/76）、72.4％（55/76）、72.4％（ 55/76）、86.8％（ 66/76）、55.3％ （42/76）,均显著高于对应癌旁正常组织[6.6％（5/76）、3.9％ （3/76）、3.9％ （3/76）、3.9％ （3/76）、2.6％（ 2/76）,x2值分别为29.01、75.39、75.39、105.34、57.18,P均＜0.001].食管鳞癌组术前血浆中5个基因的甲基化阳性率分别为42.1％（32/76）、63.2％ （48/76）、63.2％ （48/76）、71.1％ （54/76）、50.0％（ 38/76）,均显著高于健康对照组[3.3％ （2/60）、3.3％（2/60）、1.7％（ 1/60）、3.3％ （2/60）、1.7％ （1/60）,x2值分别为26.88、51.62、55.01、63.48、38.30,P均＜0.001].食管鳞癌患者血浆与肿瘤组织中5个基因甲基化的符合率均较高（Kappa值分别为0.679、0.791、0.791、0.5Objective To investigate the methylation status of multiple genes in plasma and tumor tissues and its application in molecular diagnosis of esophageal squamous cell carcinoma （ESCC）.Methods Methylation specific polymerase chain reaction （MSP） was used to detect methylation status of 5 tumor suppressor genes,such as adenomatous polyposis coli （ APC ）,retinoic acid receptor-beta2 （ RARβ2 ）,E-cadherin （CDH1）,cyclin-dependent kinase inhibitor4A （p16INK4α） and ras association domain family member 1 A （RASSF1A） in tumour tissues,adjacent normal tissues and plasma which obtained 1 d preoperative and 7 d postoperative in 76 cases with ESCC.60 healthy volunteers were randomly selected as a control which were age-matched and sex-matched.Chi square test was used to analyze DNA methylation rates of 5 genes in various groups of tissue and plasma samples; Kappa test was used to compare the consistency of DNA methylation in the plasma samples and tissue samples,and their correlation was analyzed by Spearman correlation test; Receiver operating characteristic curve （ROC） was used to evaluate the sensitivity and specificity for single gene detection and 5 genes joint detection for diagnosis of esophageal squamous cell carcinoma.Results The methylation rates of APC,RARβ2,CDH1,p16INK4α and RASSF1A in tumour tissues of patients with ESCC were 44.7% （ 34/76）,72.4% （ 55/76 ）,72.4% （55/76）,86.8% （ 66/76 ）,55.3% （42/76）,respectively,which were significantly higher than that in the corresponding adjacent normal tissues [ 6.6% （ 5/76 ）,3.9% （ 3/76 ）,3.9% （ 3/76 ）,3.9% （ 3/76 ）,2.6% （ 2/76 ）,x2 =29.01,75.39,75.39,105.34,57.18,all P 〈 0.001 ].The methylation rates of above 5 genes in patients＇ plasma were 42.1% （ 32/76 ）,63.2% （ 48/76 ）,63.2% （ 48/76 ）,71.1% （ 54/76 ）,50.0% （ 38/76 ）,respectively,which were significantly higher than that of control group [3.3% （2/60）,3.3% （2/60）,1.7% （ 1/60）,3.3% （2/60）,1.7% （1/60）,x2 =26.

关 键 词：食管肿瘤 癌 鳞状细胞 DNA甲基化 

分 类 号：R735.1[医药卫生—肿瘤]