乳腺癌循环肿瘤细胞分离鉴定试剂盒的建立  

Establishment on isolation and identification kit for circulating tumor cells in breast cancer

作　　者：张健 冯晓燕[2] 孙亭亭 王凯[4] 张希锐 高杰锋 梁晓飞[4] 康向东[5] 彭俊杰[6] 张贺秋[2] 苗朝良 沈鹤柏 

机构地区：[1]上海市临床检验中心,上海2001262 [2]军事医学科学院基础医学研究所生物诊断研究室,北京100850 [3]上海柏慧康生物科技有限公司,上海201900 [4]上海交通大学医学院附属仁济医院上海市肿瘤研究所,上海200032 [5]上海市普陀区中心医院检验科,上海200062 [6]复旦大学附属肿瘤医院大肠科,上海200032 [7]聊城市肿瘤医院,山东聊城252000

出　　处：《检验医学》2015年第10期1011-1016,共6页Laboratory Medicine

基　　金：上海市科委生物医药产业处产学研项目(12DZ1941702);上海市中小企业技术创新基金资助项目(1302H159400);第2届中国创新创业大赛上海张江扶持资金资助项目

摘　　要：目的探讨乳腺癌循环肿瘤细胞(CTC)分离鉴定试剂盒中上皮细胞黏附分子(Ep CAM)免疫脂质磁球及人乳腺珠蛋白(h MAM)单克隆抗体在乳腺癌CTC分离鉴定中的应用价值。方法将乳腺癌MCF-7细胞混于健康志愿者的新鲜血样中,采用免疫磁球分离技术分析Ep CAM免疫脂质磁球对乳腺癌细胞的分离效果。在强生Cell TracksAuto PrepCTC检测仪(简称强生CellsearchTM系统)上进行Ep CAM免疫脂质磁球和强生磁球对MCF-7细胞捕获效率的模拟实验及乳腺癌患者血液的临床检测。结果荧光显微镜的观察结果显示Ep CAM免疫脂质磁球的细胞捕获能力明显强于无抗体修饰的纳米磁球。激光共聚焦显微镜的观察结果显示Ep CAM抗体修饰可显著增强磁球捕获MCF-7细胞的效率和特异性。模拟实验结果显示Ep CAM免疫脂质磁球对10、20、40、80、150、200个MCF-7细胞的捕获结果与强生磁球接近;对乳腺癌患者血液中的CTC捕获效率与强生磁球相当,部分病例要明显优于强生磁球。结论构建的基于Ep CAM免疫脂质磁球和h MAM单克隆抗体的CTC分离鉴定试剂盒对乳腺癌患者血液中的CTC具有较高的捕获效率,这将为实现乳腺癌早期诊断、术前、术后分析及治疗效果检测提供了有力的支撑。Objective To investigate the clinical application significance of epithelial cell adhesion molecule（ Ep CAM） lipid immune magnetic particles and human breast mammaglobin（ h MAM） monoclonal antibody in breast cancer circulating tumor cell（ CTC） isolation and identification kit. Methods Breast cancer MCF-7 cells were mixed in healthy fresh blood samples,and the isolation ability of Ep CAM lipid immune magnetic particles on breast cancer was analyzed by immune magnetic particle isolating technique. A comparison between Ep CAM lipid immune magnetic particles and Cell TracksAuto PrepSystem（ Cell SearchTM） magnetic particles for the capture efficiency of MCF-7cells was performed in a simulation experiment,and the hematological items were determined. Results The results of fluorescence microscopy showed that the capture ability of Ep CAM lipid immune magnetic particle was significantly stronger than that of nanoparticles without antibody. The results of laser scanning confocal microscopy showed that the modification of Ep CAM lipid immune magnetic particle antibody significantly enhanced the efficiency and specificity of magnetic particles capturing MCF-7 cells. Simulation experiment results showed that the capturing results of Ep CAM lipid immune magnetic particles of 10,20,40,80,150 and 200 MCF-7 cells were close to Cell SearchTMmagnetic particles. In the blood samples from patients with breast cancer,the CTC capturing efficiencies of Ep CAM lipid immune magnetic particles and Cell SearchTMmagnetic particles were almost the same,in some cases,the CTC capturing efficiency of Ep CAM lipid immune magnetic particles was significantly better than that of Cell SearchTMmagnetic particles. Conclusions The research on the establishment of Ep CAM lipid immune magnetic particles and h MAN monoclonal antibody based on CTC isolation and identification kit has high capturing efficiency of CTC in blood samples from patients with breast cancer. It will provide a strong support for the early diagnosis,pre-operati

关 键 词：循环肿瘤细胞 上皮细胞黏附分子 免疫磁球 人乳腺珠蛋白 

分 类 号：R446.1[医药卫生—诊断学]