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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 a method for the detection of circulating tumor cells (CTCs) by membrane filtration separation of tumor cells (ISET) and laser scanning cytometry (LSC). Methods Different numbers of breast cancer cell lines (MCF-7) were respectively mixed into healthy blood samples. ISET combined with LSC scanning was used to establish a stable detection method. The recovery rate, sensitivity and specificity of this method were also verified Sex. Results The method for detecting CTC by using ISET (8 μm pore size polycarbonate membrane) in combination with LSC and using CK8 antibody and CD45 antibody as the positive and negative selection markers of tumor epithelial cells was established. The recovery rate of tumor cells was ( 10.34 ± 1.15)%, the recovery rate remained stable in the range of 60 to 600000 tumor cells (P> 0.05), and the sensitivity of one tumor cell in 1ml blood could be detected. In the same conditions, the blank control group was not detected Tumor cells. Conclusion ISET combined with LSC detection of malignant tumors in patients with CTC method is simple, sensitive, specific and is expected to be further applied to clinical.