实时荧光定量PCR法检测乳腺癌外周血循环肿瘤细胞的临床价值

The clinical value of detecting circulating tumor cells in peripheral blood of patients with breast cancer by real time QPCR

目的:比较运用实时荧光定量PCR法联合不同生物标志物细胞角蛋白19(cytokeratin 19,CK19)、乳珠蛋白(humanmammaglobin,hMAM)、小黏蛋白(small breast epithelial mucin,SBEM)检测乳腺癌外周血循环肿瘤细胞(circulating tumor cells,CTCs)的临床价值。方法:运用实时荧光定量PCR法检测人乳腺癌细胞株MCF-7中的生物标志物CK19、hMAM、SBEM的表达情况,对20例健康志愿者和20例乳腺良性疾病患者(对照组人群)进行检测以确定CK19、hMAM、SBEM在外周血中的基础表达水平,检测94例可手术乳腺癌患者外周血中CK19、hMAM、SBEM表达情况。结果:人乳腺癌细胞株MCF-7中CK19、hMAM、SBEM均有表达,Ct值分别为13.87、34.47、20.14;40例对照组人群中有1例表达CK19,另有1例表达hMAM,未检测到SBEM表达;94例可手术乳腺癌患者中CK19表达率49%、hMAM表达率34%、SBEM表达率13%,联合CK19、hMAM、SBEM阳性率为56%。结论:运用多标志物(CK19、hMAM、SBEM)联合的实时荧光定量PCR法检测乳腺癌外周血CTCs具有较高的敏感性,同时特异性并不降低。

Objective： To compare clinical value of different biomarkers （CK19,hMAM,SBEM） used to detect circulating tumor cells（CTCs） in peripheral blood of patients with breast cancer using real-time quantitative PCR （QPCR） platform. Methods： The expression of CKI9,hMAM,SBEM in human breast cancer cell lines MCF-7 was detected by real time QPCR,peripheral blood samples of 20 healthy volunteers and 20 patients with breast benigh disease were detected to identify the base line expression of CK19, hMAM,SBEM. The expression of CK19,hMAM,SBEM in peripheral blood from 94 patients with operable breast cancer was detected also. Results： CK19,hMAM and SBEM were all detected in MCF-7,and the Ct value were 13.87,34.47,and 20.14,respectively;a- mong the 40 control persons,one sample expressed CK19,one sample expressed hMAM,and no sample expressed SBEM;among the 94 patients with operable breast cancer,46 （49%） samples expressed CK19,32 （34%） samples expressed hMAM, and 12（13%） sampies expressed SBEM, 53 （56%） samples were positive by combined detection using the three biomarkers. Conclusion： It is higher sensitivity to detect circulating tumor cells in peripheral blood of breast cancer combined detection using muhimarkers （CK19, hMAM, SBEM） based on a real time QPCR platform,yet specificity was not depressed.