定量PCR检测肝细胞癌患者血浆循环DNA及其临床意义  

Quantitation of plasma circulating DNA in hepatocellular carcinoma using quantitative PCR and its potential clinical value

作　　者：黄朝晖[1] 胡瑜[1] 华东[1] 程之红[1] 谢其根[1] 王琼瑶[1] 王丰[1] 周锡科[1] 杜祥[2] 

机构地区：[1]苏州大学附属第四医院肿瘤研究所,江苏无锡214062 [2]复旦大学附属肿瘤医院病理科,复旦大学上海医学院肿瘤学系,上海200032

出　　处：《中国癌症杂志》2010年第9期663-667,共5页China Oncology

基　　金：江苏省自然科学基金资助项目(项目编号:BK2008114)

摘　　要：背景与目的:血循环DNA是一种新的肿瘤诊断及预后判断的标志物。本研究运用定量PCR技术检测肝细胞癌(hepatocellular carcinoma,HCC)患者血浆循环DNA含量并探讨其诊断价值。方法:收集72例HCC患者术前血浆样本,37例肝良性病变(肝硬化以及慢性肝炎)和41例健康志愿者的血浆样本,纯化血浆循环DNA,采用实时定量PCR技术对血浆DNA水平进行检测。应用接受者操作特性(receiver-operating characteristics,ROC)曲线分析血浆循环DNA在HCC诊断中的价值。结果:HCC中位血浆循环DNA浓度(173ng/mL)显著高于健康对照(9ng/mL)和肝良性病变组(46ng/mL)(P<0.001);其ROC曲线下面积(area under the ROC curve,AUC)分别为0.949和0.874。而HCC血浆DNA浓度也显著高于肝硬化及慢性肝炎患者(P=0.001),AUC为0.703。以18.2ng/mL作为诊断HCC的临界值,其诊断特异度为90.2%,敏感度达90.3%;与血清AFP联合检测可提高HCC诊断效率,AUC上升至0.974,其诊断特异度和敏感度分别为95.1%和94.4%。伴肝内播散或脉管癌栓HCC患者的血浆DNA浓度(261ng/mL)明显高于不伴肝内播散灶或脉管癌栓患者(142ng/mL,P=0.035)。结论:定量PCR技术可精确定量血浆循环DNA浓度;血浆DNA分析对于HCC诊断,预测肿瘤转移潜能具有重要价值。Background and purpose：Circulating DNA is a potential biomarker for tumor diagnosis and prognosis.This study aimed to quantify circulating DNA in plasma from patients with hepatocellular carcinoma （HCC） by using the quantitative PCR method.The data would be evaluated for its potential clinical value.Methods：Plasma samples were collected from 72 HCC patients,37 patients with liver cirrhosis or chronic hepatitis and 41 healthy volunteers.Plasma DNA was extracted and quantifi ed using a real-time quantitative PCR method.Receiver-operation characteristic （ROC） curves were plotted to measure the overall effi ciency of this method in HCC diagnosis.Results：DNA concentration in the HCC plasma （median：173 ng/mL） was signif icantly higher than that in the healthy controls （9 ng/mL） or benign control patients （46 ng/mL） （P0.001）.The area under the ROC curve （AUC） assessing plasma DNA was 0.949 for healthy controls and 0.874 among control patients.Plasma DNA detection could discriminate HCC from normal controls with a 90.2% sensitivity and 90.3% specifi city with a cut-off value of 18.2 ng/mL.Combined ROC analyses using plasma DNA and AFP revealed an elevated AUC of 0.974 with a 95.1% sensitivity and 94.4% specifi city in discriminating HCC from normal controls.The plasma DNA in HCC patients with intrahepatic spreading or vascular invasion （261 ng/mL） was signifi cantly higher than that in those without intrahepatic spreading or vascular invasion （142 ng/mL） （P=0.035）.Conclusion：Quantitative PCR could detect plasma DNA effi ciently.Plasma DNA may be a valuable noninvasive diagnostic tool for detecting and predicting the metastatic potential of HCC.

关 键 词：肝细胞癌 血浆 定量PCR 循环DNA 

分 类 号：R735.7[医药卫生—肿瘤]