高灵敏度hMAM-QDs脂质体结合CK19双标记法检测乳腺癌循环肿瘤细胞的临床研究

目的:评估细胞角蛋白19（CK19）单标记法和CK19结合人乳球蛋白-量子点（hMAM-QDs）双标记法检测乳腺癌循环肿瘤细胞（circulating tumor cells，CTCs）的临床价值。方法:纳米脂质体（PL）技术用于构建hMAM-PL（QDs）；借助免疫磁珠分离技术，使用上皮细胞黏附分子（EpCAM）抗体分离乳腺癌CTCs。结果:hMAM-PL（QDs）［平均粒径（102.6±1.1）nm，多分散指数0.047］可以显著提高实验中的检测灵敏度。此外，CK19-FITC与hMAM-PL（QDs）双标记方法相结合可有效区分分离的CTCs，提高准确性，并减少乳腺癌检测中的假阳性。CTCs编号、图像与临床特征以及病理状况高度一致。结论:CK19结合hMAM-PL（QDs）双标记方法检测比CK19单标记方法更准确。双标记方法可以显著减少假阳性结果，对于早期乳腺癌的诊断、治疗和预后具有重要意义。

High-sensitivity hMAM-QDs liposomes combined with CK19 double-labeled method detection for circulating tumor cells in breast cancer and early clinical verification

Objective:To evaluate the clinical value of detecting circulating tumor cells (CTCs) in breast cancer by cytokeratin 19 (CK19) single-labeled method and CK19 combined with human mammaglobin-quantum dots (hMAM-QDs) double-labeled method.Methods:Nanopolymeric liposome (PL) technology was used to build hMAM-PL (QDs),and epithelial cell adhesion molecule (EpCAM) antibody immune magnetic ball separation technology was applied to separate breast CTCs.Results:hMAM-PL (QDs) ［average particle size (102.6±1.1)nm with polydispersity index 0.047］ can obviously improve detection sensitivity in cell experiments.Furthermore,CK19-FITC combined with hMAM-PL (QDs) double-labeled method can efficiently distinguish separated CTCs,increase accuracy,and decrease false positive in breast cancer detection.The CTCs numbers and images were highly consistent with clinical characteristics and pathology conditions.Conclusion:The CK19 combined with hMAM-PL (QDs) double-labeled method detection is more accurate than the CK19 single-labeled method.The double-labeled method can significantly decrease false positive results,and is useful for early breast cancer diagnosis,treatment and prognosis.