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目的应用广谱抗细胞角蛋白(Cytokeratin,CK)抗体CK3-6H5检测上皮组织来源的肿瘤细胞系中CK的表达,并研究应用此抗体检测血液循环中此类肿瘤细胞的敏感性。方法应用CK3-6H5标记QGY-7703等26种人类上皮组织来源的肿瘤细胞系、人Burkitt淋巴瘤细胞系Raji、人T淋巴细胞白血病细胞株Jurkat以及健康志愿者外周血单个核细胞,通过免疫荧光显色和免疫细胞化学技术检测其中CK的表达情况;以表达CK的人肝癌细胞系QGY-7703作为靶细胞,以不同的比例分别掺入到健康志愿者PBMC中,以免疫磁珠去除CD45+的白细胞,对肿瘤细胞进行富集,流式细胞仪检测其中CK+CD45-肿瘤细胞数目,分析用CK3-6H5检测外周血循环肿瘤细胞(circulating tumor cell,CTC)的敏感性。结果所有上皮组织来源肿瘤细胞系均表达CK,而健康志愿者和淋巴瘤细胞系Raji、T淋巴细胞白血病细胞株中未检测到CK表达。当向2×108个PBMC中掺入20个QGY-7703细胞时,经免疫磁珠富集后,行FACS分析,仍能检测到12个肿瘤细胞。结论CK3-6H5抗体复合物可以识别常见的上皮来源的肿瘤,应用CK3-6H5抗体标记肿瘤细胞,采用免疫磁珠富集技术以及流式细胞技术,可以用于有效检测外周血中上皮来源的肿瘤细胞。
Objective To detect the expression of cytokeratin (CK) CK3-6H5 in epithelial-derived tumor cell lines and to study the sensitivity of this antibody to detect such tumor cells in the blood circulation. Methods Tumor cell lines derived from 26 human epithelial tissues, such as QGY-7703, labeled with CK3-6H5, Raji human Burkitt lymphoma cell line, Jurkat human leukemia cell line Jurkat, and peripheral blood mononuclear cells from healthy volunteers were detected by immunofluorescence The colorimetric and immunocytochemical techniques were used to detect the expression of CK. The human hepatocarcinoma cell line QGY-7703 expressing CK was used as target cells and spiked into PBMCs of healthy volunteers in different proportions. The immunomagnetic beads were used to remove CD45 + Leukocytes, and enrich the tumor cells. The number of CK + CD45- tumor cells was detected by flow cytometry. The sensitivity of CK3-6H5 for detecting circulating tumor cells (CTCs) was analyzed. Results All epithelial-derived tumor cell lines expressed CK, but CK expression was not detected in Raji and T-lymphoblastic leukemia cell lines from healthy volunteers and lymphoma cell lines. When 20 QGY-7703 cells were spiked into 2 × 108 PBMCs, 12 tumor cells could still be detected by FACS analysis after enrichment with immunomagnetic beads. Conclusions The CK3-6H5 antibody complex can recognize common epithelial tumor. Using CK3-6H5 antibody to label tumor cells, immunomagnetic beads enrichment technique and flow cytometry can be used to detect epithelial-derived tumors in peripheral blood cell.