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目的建立基于细胞学水平的循环肿瘤细胞负性筛选方法,检测食管鳞癌患者外周血循环肿瘤细胞。方法选用负性筛选策略,分步去除7.5ml食管鳞癌外周血样本中的红、白细胞,富集上皮来源的稀有细胞。比较3种免疫磁珠(Miltenyi、AVIVA和Xinzhi磁珠)去除白细胞、富集外周血循环肿瘤细胞的特点和效率。用经典的细胞化学染色鉴定循环肿瘤细胞。结果Miltenyi和AVIVA磁珠去除白细胞、富集肿瘤细胞效果较好。从富集后的食管鳞癌患者外周血稀有细胞中鉴定到形态各异的循环肿瘤细胞。结论负性筛选策略可以有效分离鉴定食管鳞癌患者外周血形态各异的循环肿瘤细胞。
Objective To establish a negative screening method of circulating tumor cells based on cytology and to detect the circulating tumor cells in patients with esophageal squamous cell carcinoma. Methods Negative screening strategy was used to remove the red and white blood cells from 7.5 ml of esophageal squamous cell carcinoma in stages and enriched the epithelial derived rare cells. Comparison of three kinds of immunomagnetic beads (Miltenyi, AVIVA and Xinzhi beads) to remove leukocytes and enrich the characteristics and efficiency of peripheral blood circulating tumor cells. Cyclic tumor cells are identified by classical cytochemical staining. Results Miltenyi and AVIVA beads removed leukocytes and enriched tumor cells. From the enriched esophageal squamous cell carcinoma patients with peripheral blood cells identified in different forms of circulating tumor cells. Conclusion Negative screening strategy can effectively separate and identify circulating tumor cells with different morphological features in patients with esophageal squamous cell carcinoma.