流式细胞仪检测非小细胞肺癌患者活性循环内皮细胞初探

An introduction to flow cytometric analysis for measuring activated cyclic endothelial cells in the peripheral blood of NSCLC patients

目的建立流式细胞仪技术检测成熟循环内皮细胞(CECs)及活性循环内皮细胞(aCECs)方法,了解非小细胞肺癌(NSCLC)患者及健康人群的CECs和aCECs数量上的差异。方法采用淋巴细胞分离技术提取外周血单个核细胞,用CD45-PC5、CD146-PE、CD106-FITC抗体标记60例肺癌患者(NSCLC 56例)及30例健康人,用流式细胞仪检测CECs及aCECs。结果采用上述标记抗体通过流式细胞仪可顺利检测出外周血CECs及aCECs,而NSCLC患者外周血CECs及aCECs的数量[(2 573±2 019)个/mL,(784±914)个/mL]明显高于健康人群[(273±210)个/mL,(35±61)个/mL],差异有统计学意义(P<0.001)。aCECs仅占CECs的一小部分,NSCLC患者中aCECs占CECs的比例明显高于健康人群(P<0.001),差异有统计学意义。在不同病理类型肺癌及临床分期上CECsa、CECs的数量无统计学差异(P>0.05)。CECs、aCECs的数量与NSCLC肿瘤大小、血CEA之间无明显相关性(P>0.05)。结论采用流式细胞仪可以快速、方便检测出CECs及aCECs。CECs及aCECs的数量在NSCLC患者外周血均有明显升高,它可以作为反映NSCLC的血管生成状况及抗血管生成治疗疗效的标志物。

Objective To establish the flow cytometric method for circulating endothelial cells（CECs） and activated circulating endothelial cells（aCECs）,for finding out the quantitative difference of CECs and aCECs between NSCLC patients and healthy controls.Methods We adopted the method of Lymphocyte separation to prepare mononuclear cell samples;CECs and aCECs were enumerated in 60 lung cancer patients（56 NSCLC） and 30 healthy controls by flow cytometry using PC5-labelled anti-CD45,FITC-labelled anti-CD106 and PE-labelled anti-CD146.Results Adopting the above method can successfully detect the numbers of CECs and aCECs.The counts of CECs and aCECs were significantly higher in NSCLC patients[（2 573±2 019）/mL,（784±914）/mL] than in healthy volunteers[（273±210）/mL,（35±61）/mL]（P0.001）.CECs was onlya smallpart of aCECs;aCECs percent was higher in NSCLC patients than that in healthy volunteers（P0.001）.There was no difference in the amounts of CECs and aCECs in terms of pathology type and clinical stage in lung cancer patients,and we did not find the correlation between the CECs and aCECs count and tumor sizes,as well as blood CEA（P0.05）.Conclusion Adopting the above method can rapidly and conveniently detect the number of CECs and aCECs.significantly increased CECs and aCECs in NSCLC patients can be surrogate biomarkers of angiogenesis and antiangiogenic drug activity.