用膜滤过联合激光扫描细胞仪检测恶性肿瘤患者循环肿瘤细胞

Detection of Circulating Tumor Cells with Cell Filtration and Laser Scanning Cytometry

目的建立用膜滤过分离肿瘤细胞技术(ISET)联合激光扫描细胞仪(LSC)检测循环肿瘤细胞(CTC)的方法。方法将不同数量的乳腺癌细胞系(MCF-7)细胞分别混入健康血标本中,用ISET联合LSC自动扫描分析,以建立稳定的检测方法,同时验证该方法肿瘤细胞回收率、敏感性及特异性。结果建立了用ISET(用8μm孔径的聚碳酸酯滤膜)联合LSC,并用CK8抗体、CD45抗体分别作为肿瘤上皮细胞的阳性和阴性选择标记来检测CTC的方法;该方法肿瘤细胞回收率为(10.34±1.15)%,在混入60～600000个肿瘤细胞范围内回收率保持稳定(P>0.05);达到可检测到1ml血中一个肿瘤细胞的敏感性,相同条件下空白对照组均未检出肿瘤细胞。结论用ISET联合LSC检测恶性肿瘤患者CTC方法简便、敏感性高、特异性强,有望进一步应用于临床。

Objective To establish the method of cell filtration（isolated by the size of epithelial tumor cells,ISET）combined with laser scanning cytometry（LSC）to detect circulating tumor cells（CTC）in cancer patients.Methods Tumor cells（MCF-7）were mixed with the blood samples from healthy volunteers at different ratios.ISET and LSC were used to detect tumor cells.The rate of recovery,sensitivity and specificity for this detection method were determined.Results The repeated sensitivity assays confirmed that the recovery rate of mixed tumor cells with this method remained about（10.34±1.15）% and the result kept stable when mixed cells was added at a number of between 60 and 600 000（P0.05）.One tumor cell in 1 ml peripheral blood would be detected while no positive cells were detected in the controls,indicating the high sensitivity and specificity of the method.Conclusion The method with cell filtration and laser scanning cytometry for CTC detection shows with high sensitivity and specificity,which can be applied in the studies of relationships between CTC and tumor progression.