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目的 运用流式细胞仪定量分析外周血循环中肺癌细胞。方法 外周血经Ficoll梯度离心分离单核细胞组分,后者用CD45、细胞角蛋白(CK)及肺癌特异性单抗(2F7/S5A)染色后,应用流式细胞仪检测CD45- CK+ 2F7/S5A+ 细胞。然后计算每升血中上述细胞含量。结果 (1)外周血单核细胞的表型为CD45+ CK- 2F7/S5A- ,而小细胞和非小细胞肺癌细胞系均为CD45- CK+ 2F7/S5A+ 。因此,应用此方法能从106 白细胞中检测到1个肺癌细胞,敏感性达到每毫升血中检测到5 个肺癌细胞;(2)15 例正常人及7 例肺部良性疾患外周血中CD45- CK+ 2F7/S5A+ 细胞均为阴性,而40例肺癌患者外周血14 例发现阳性细胞,阳性率达35 % ,阳性细胞含量平均为1×105 /L。结论 本方法具有较好的敏感性和特异性,能够定量分析外周血中的微量肺癌细胞
Objective To quantitatively analyze lung cancer cells in peripheral blood circulation by flow cytometry. Methods Peripheral blood was separated by Ficoll gradient centrifugation. The latter was stained with CD45, cytokeratin (CK) and lung cancer-specific monoclonal antibody (2F7/S5A). CD45-CK+ 2F7/ was detected by flow cytometry. S5A+ cells. Then calculate the above cell content per liter of blood. Results (1) The phenotype of peripheral blood mononuclear cells was CD45+ CK-2F7/S5A-, while both small cell and non-small cell lung cancer cell lines were CD45-CK+ 2F7/S5A+. Therefore, using this method, one lung cancer cell can be detected from 106 leukocytes, with a sensitivity of 5 lung cancer cells detected per milliliter of blood; (2) CD45- in the peripheral blood of 15 normal individuals and 7 benign lung diseases. CK+ 2F7/S5A+ cells were negative, and 40 cases of lung cancer patients found positive cells in 14 cases, the positive rate was 35%, the average positive cell content was 1×105/L. Conclusion This method has good sensitivity and specificity and can quantitatively analyze trace lung cancer cells in peripheral blood.