【摘要】 目的:建立一种荧光定量PCR方法用于血浆循环DNA定量,并讨论血浆循环DNA检测在乳腺癌诊断及预后中的意义。方法:以61例乳腺癌患者、33例乳腺良性病变患者和25例健康对照者血浆为材料,用Qiagen纯化柱提取血浆DNA,采用荧光定量PCR技术(SYBR Green I染料法)对血浆DNA水平进行检测,DNA水平用临界域值时的循环数(Ct值)表示,并应用ROC曲线分析血浆循环DNA定量在乳腺癌诊断中的价值。结果:乳腺癌血浆循环DNA水平(27.71±1.41)显著高于健康对照(30.37±1.32)和良性对照(29.7±1.12)(P<0.001),ROC曲线下面积分别为0.921和0.837。循环DNA水平与分期、淋巴结转移、肿瘤大小、年龄及雌激素受体状况均无相关性(P>0.05)。术后乳腺癌患者血浆循环DNA水平显著下降,其中复发患者血浆循环DNA水平显著高于未复发者。结论:血浆循环DNA水平对于乳腺癌诊断和预后均具有一定临床意义。更多还原

【Abstract】 Objective:To develop a real-time florescence quantitative polymerase chain reaction(PCR) method for quantitation of circulating DNA in plasma and explore its value in the diagnosis and prognosis of breast cancer.Methods:Plasma DNA from 119 women(61 patients with breast cancer,33 control patients with benign breast diseases and 25 healthy control) extracted using a Qiamp blood mini kit and was quantified by a real-time PCR method(SYBR Green I dye assay),the DNA concentration was expressed as cycle threshold(Ct).Receiver-operation characteristic(ROC) curves were plotted to measure the overall ability of this method in discriminating breast cancer from the healthy and benign breast disease.Results:DNA concentration in the plasma of the breast cancer patients(27.71±1.41) was significantly higher than that in the control patients(29.7±1.12) or healthy control(30.37±1.32)(P<0.001)..The area under the ROC curve assessing plasma DNA concentration was 0.917 for healthy controls and 0.837 for control patients.No association was observed between plasma DNA levels and tumor stage,nodal status,tumor size,pathologic stage,age,and estrogen receptor(P>0.05).The median DNA concentration in the follow-up plasma samples was significantly lower than those recorded in the matched samples before surgery.In addition,median DNA concentration at follow-up was significantly lower in disease-free individuals as compared with the patients with proven cancer relapse(P<0.01).Conclusion:Quantification of plasma circulating DNA,using real-time quantitative PCR,may be a valuable noninvasive diagnostic tool in discriminate breast cancer from healthy individuals.更多还原