Cancer Metastasis Alert and Prevention Center, and Biopharmaceutical Photocatalysis of State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou, China.
Surgery Department, Fuzhou General Hospital, Fuzhou, China.
Cancer. 2015 Sep 1;121(17):3036-45. doi: 10.1002/cncr.29444. Epub 2015 May 6.
This study was aimed at establishing a sensitive and specific isolation, characterization, and enumeration method for living circulating tumor cells (CTCs) in patients with colorectal carcinoma.
Quantitative isolation and characterization of CTCs were performed through a combination of immunomagnetic negative enrichment and fluorescence-activated cell sorting. Isolated CTCs were identified by immunofluorescence staining. The viability and purity of the sorted cells were determined by flow cytometry. Blood samples spiked with HCT116 cells (range, 3-250 cells) were used to determine specificity, recovery, and sensitivity. The method was used to enumerate, characterize, and isolate living CTCs in 10 mL of blood from patients with colorectal carcinoma.
The average recovery of HCT116 cells was 61% or more at each spiking level, and the correlation coefficient was 0.992. An analysis of samples from all 18 patients with colorectal carcinoma revealed that 94.4% were positive for CTCs with an average of 33 ± 24 CTCs per 10 mL of blood and with a diameter of 14 to 20 μm (vs 8-12 μm for lymphoma). All patients were CD47(+) , with only 4.3% to 61.2% being CD44(+) . The number of CTCs was well correlated with the patient TNM stage and could be detected in patients at an early cancer stage. The sorted cells could be recultured, and their viability was preserved.
This method provides a novel technique for highly sensitive and specific detection and isolation of CTCs in patients with colorectal carcinoma. This method complements the existing approaches for the de novo functional identification of a wide variety of CTC types. It is likely to help in predicting a patient's disease progression and potentially in selecting the appropriate treatment.
本研究旨在建立一种敏感、特异的方法，用于从结直肠癌患者循环肿瘤细胞（CTC）中进行定量分离、鉴定和计数。
采用免疫磁珠阴性富集与流式细胞分选相结合的方法，对 CTC 进行定量分离和鉴定。通过免疫荧光染色鉴定分离的 CTC。采用流式细胞术检测分选细胞的活力和纯度。用 HCT116 细胞（范围 3-250 个细胞）对血液样本进行加标，以确定特异性、回收率和灵敏度。采用该方法对 10ml 结直肠癌患者血液中的活 CTC 进行计数、鉴定和分离。
在每个加标水平，HCT116 细胞的平均回收率均在 61%以上，相关系数为 0.992。对 18 例结直肠癌患者的样本分析显示，94.4%的患者 CTC 阳性，平均每 10ml 血液中有 33±24 个 CTC，直径为 14-20μm（而淋巴瘤为 8-12μm）。所有患者均为 CD47(+)，仅有 4.3%-61.2%为 CD44(+)。CTC 数量与患者 TNM 分期密切相关，可在早期癌症阶段的患者中检测到。分选细胞可再培养，活力得以保存。
该方法为结直肠癌患者 CTC 的高敏感、特异检测和分离提供了一种新方法。该方法补充了现有方法，可用于对多种 CTC 类型进行从头鉴定。有望帮助预测患者的疾病进展，并可能有助于选择合适的治疗方法。
