负性富集食管鳞癌患者外周血中循环肿瘤细胞的检测及其意义

Circulating tumor cells from peripheral blood in esophageal squamous cell carcinoma by negative selection and its clinical significance

目的:负性富集并鉴定食管鳞癌患者外周血循环肿瘤细胞(CTCs)的数量及表型特征,探讨CTCs在食管鳞癌临床疗效评价和预后判断等方面的应用价值。方法:通过免疫磁珠偶联CD45抗体除去外周血白细胞,富集外周血CTCs的"负性策略",用荧光鉴定CTCs上皮标志及细胞核特征,用统计学方法分析CTCs数量改变与患者临床参数与治疗效果、预后的相关性。结果:59例食管鳞癌患者术前外周血CTCs的阳性率为79.7%(47/59),7.5ml外周血CTCs平均检出数目为7.83(0~72)个。其中淋巴结是否转移、肿瘤TNM分期、肿瘤浸润程度和分化程度、不同各组CTCs检出的差异,具有统计学意义(P<0.05)。采用多因素Cox回归分析,术前CTCs≥5及淋巴结转移是影响预后的显著因素。结论:免疫磁珠负性富集结合免疫荧光鉴定的方法可以有效地分离和鉴定食管癌外周血CTCs。初步研究提示,外周血中CTCs与食管鳞癌分期、进展及其预后相关。

Objective： To use the ＂negative strategy＂ of immunomagnetic beads coupled with antibody to enrich the CTCs in peripheral blood of patients with esophageal squamous cell carcinoma（ESCC） and study the quantitative and phenotypic characteristics of CTCs, to explore the value of CTCs in clinical evaluation and prognosis of ESCC. Meth- ods：The peripheral blood leukoeytes were removed by the ＂negative strategy＂ and the CTCs were enriched. The epi- thelial signs and nuclear features of CTCs were identified by fluorescence. We analyzed the correlation between the number of CTCs and clinical parameters by statistical methods. Results ： The positive rate of CTCs of 59 patients with ESCC was 79.7% （47/59） ,the average number of CTCs in 7.5 ml peripheral blood was 7.83 （0 -72）. The differ- ences of CTCs detection in different lymph node metastasis, TNM stage, tumor invasion and differentiation were statis- tically significant. Using multivariate Cox regression analysis, CTCs I〉 5 before operation and lymph node metastasis were significant factors affecting the prognosis. Conclusion：The negative enrichment of immunomagnetic beads com- bined with immunofluorescence could effectively isolate and identify CTCs of peripheral blood in ESCC. Preliminary study suggests that peripheral blood CTCs was associated with the stage, progression and prognosis of ESCC.