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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 the detection of cytokeratin 19 (CK19), human mammary carcinoma (hMAM) and small breast epithelial mucin (SBEM) by real-time fluorescence quantitative PCR in peripheral blood of breast cancer The clinical value of circulating tumor cells (CTCs). Methods: The expression of biomarkers CK19, hMAM and SBEM in human breast cancer cell line MCF-7 were detected by real-time fluorescence quantitative PCR. Twenty healthy volunteers and 20 benign breast diseases patients (control group) The levels of CK19, hMAM and SBEM in peripheral blood were detected to detect the expression of CK19, hMAM and SBEM in 94 patients with operable breast cancer. Results: The expression of CK19, hMAM and SBEM in human breast cancer cell line MCF-7 was 13.87, 34.47 and 20.14, respectively. One of the 40 control subjects expressed CK19 and the other one expressed hMAM, The expression of SBEM was detected in 94 cases of breast cancer. The expression rate of CK19 was 49%, the expression rate of hMAM was 34%, the expression rate of SBEM was 13%, and the positive rate of CK19, hMAM and SBEM was 56%. Conclusion: The detection of CTCs in peripheral blood of breast cancer patients by using multiple markers (CK19, hMAM, SBEM) in combination with real-time fluorescence quantitative PCR method is highly sensitive and does not reduce the specificity.