一种新的富集并计数肺癌脑膜转移患者脑脊液中恶性肿瘤细胞的方法

A novel methodology to concentrate and quantify malignant cells in CSF from lung cancer patients with leptomeningeal metastasis

目的：通过观察肿瘤标志物免疫荧光染色-染色体荧光原位杂交技术平台（TM-iFISH）富集并计数肺癌脑膜转移患者脑脊液中的恶性肿瘤细胞，探讨一种新的检测脑脊液中恶性肿瘤细胞的方法。方法选取6例经脑脊液细胞学或头增强MRI扫描确诊的肺癌脑膜转移患者，每例患者经腰椎穿刺获取脑脊液20 mL（共10份），其中7.5 mL应用TM-iFISH技术富集并计数脑脊液中的恶性肿瘤细胞，10 mL行脑脊液细胞学检查，2.5 mL行脑脊液生物化学检查。结果10份脑脊液标本均顺利完成上述检测，其中7份标本通过TM-iFISH技术计数示肿瘤细胞数为3～1823个/7.5 mL脑脊液，3份脑脊液细胞学检查发现肿瘤细胞，9份脑脊液生物化学检查结果蛋白量高于正常值。3例患者治疗后应用TM-iFISH技术再次计数脑脊液恶性肿瘤细胞，2例患者计数较治疗前减少。结论 TM-iFISH技术可以富集并计数肺癌脑膜转移患者脑脊液中恶性肿瘤细胞，可能成为肺癌脑膜转移的诊断和疗效评价方法。

Objective To assess the value of tumor marker immunostaining-FISH （TM-iFISH） technology on concen？trating and enumeration of tumor cells in CSF of lung cancer patients with leptomeningeal metastasis（LM）. Methods Six cases of non-small cell lung cancer with leptomeningeal metastasis, which were diagnosed by CSF cytology or enhanced MRI scan, were selected. A total of 20 mL of CSF was collected in each case. TM-iFISH technology was employed to concen？trate and quantify circulating tumor cells in 7.5 mL CSF samples in each case while CSF cytology used 10 mL CSF samples in each case;Finally, the rest 2.5 mL CSF in each case was used for biochemistry assay. Results Ten CSF samples from 6 patients with non-small lung cancer with LM were assayed and tumor cells numbers ranging between 3 and 1 823 every 7.5 mL were found in 7 samples. On the other hand, CSF cytology examination only revealed tumor cells in 3 cases. Using CSF biochemical assay, higher than normal of protein level was found in 9 cases. TM-iFISH technology was employed again in 3 cases of patients who received treatment. Tumor cell count in CSF reduced in 2 out of the 3 cases. Conclusion TM-iFISH technology is a new method for detection and enumeration of tumor cells in the CSF in non-small cell lung cancer patients with leptomeningeal metastasis. This technology present diagnosis and curative values in lung cancer patients with leptomen？ingeal metastasis.