磁激活细胞分选技术用于结直肠癌外周血癌细胞富集的研究

The study of enriching peripheral circulating carcinoma cells in colorectal cancer patients by magnetic activated cell sorting

目的用磁激活细胞分选技术富集并定量分析结直肠癌患者外周血循环中的癌细胞。方法术前抽取 2 1例结直肠癌患者和 9例对照者的外周血样 ,标记带磁珠的细胞角蛋白 (Cytokeratin ,CK)抗体 ,经磁柱富集CK+ 上皮细胞 ,用流式细胞仪检测其含量。结果在未经富集的标本中均未发现CK+ CD45 -细胞 ;在经富集的标本中其含量在对照组与患者组间 (P <0 0 1)以及DukesA B期与DukesC D期患者间 (P <0 0 1)均差异有显著意义。结论本方法能有效地富集上皮来源细胞 ;CK+ CD45 -细胞能反映外周组织中肿瘤细胞情况 ;其与恶性肿瘤的存在。

ObjectiveTo enrich and quantitatively analyze peripheral circulative carcinoma cells by magnetic activated cell sorting (MACS) in colorectal cancer patients.MethodsBlood samples, preoperatively collected from 21 patients with colorectal cancer and 9 healthy volunteers, were labeled by anti-cytokeratin (CK) with magnetic microbeads, and passed through magnetic columns; CK +carcinoma cells were separated from the samples, and quantified by flow cytometry.ResultsCK +CD45 -cells could not be detected from the samples without MACS; in the sample undergoing MACS, the concentrations of CK +CD45 -cells were significantly higher in patient group than in control group(P<0.01), and lower in Dukes A-B patients than in Dukes C-D patients(P<0.01).ConclusionsMACS can effectively enrich peripheral epithelial tumor cells. The close relationships can be found between CK +CD45 -cell concentrations and tumor existence or clinical stages. The technique could become a valuable tool to guide evaluation of cancer metastasis risk, prognosis and chemical therapy.