应用广谱抗体CK3-6H5检测细胞角蛋白在人类上皮组织来源的肿瘤细胞系中的表达  

Use of a pan-cytokeratin antibody CK3-6 H5 for the detection of cytokeratin to detect the expressions in human epithelia-deriverd tumor cell lines

作　　者：于鑫[1] 魏玉华[1] 彭吉润[1] 郭立民[1] 赵丽君[2] 公磊[1] 王卉 李澍[1] 冷希圣[1] 

机构地区：[1]北京大学人民医院肝胆外科中心,北京100044 [2]北京大学人民医院妇科,北京100044 [3]德国美天旎生物技术有限公司

出　　处：《中国实验诊断学》2007年第8期995-998,共4页Chinese Journal of Laboratory Diagnosis

基　　金：北京大学人民医院研究与发展基金(RDB2006-12)

摘　　要：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 investigate the expression of eytokeratin （CK） in epithelia-deriverd tumor cell lines by pan-eytokeratin antibody CK3-6H5 and to estimate its sensitivity to detect circulating epithelia-defiverd tumor cells. Methods Marked by CK3-6H5, the CK expression in twenty-six epithelia-deriverd tumor cell lines including human hepatocellular carcinoma cell line QGY-7703 were detected by immunofluoreseenee and immumocytochemistry, with human Burkitt lymphoma cell line Raji, human T lymphocyte-derived leukemia cell line Jurkat and peripheral blood mononuclear Cells （PBMCs） from healthy volunteers as controls. In order to estimate the sensitivity to detect circulating epithelia-defiverd tumor cells using CK3-6H5, CK-positive QGY-7703 cells were diluted into PBMCs from healthy volunteers according to different ratios, after CD45 ^＊ white cells were depleted by immunomagnectie beads, enriched tumor cells were analysed by FACS. Results All the twenty-six epithelia-deriverd tumor cell lines were CK positive,whereas no CK expression were detected in cell line Raji, Jurkat and PBMCs from healthy volunteers. Twenty QGY-7703 cells were diluted into 2 × 108 PBMCs from healthy volunteers, twelve tumor cells could still be detected by FACS after the enrichment by immunomagnectie beads.Conclusion epithelia-deriverd tumor cells could be identified by pan-eytokeratin antibody CK3-6H5,when marked these tumors with CK3-6H5, the circulating epithelia-deriverd tumor cells could be effectively detected by combing immunomagnectic beads enrichment and FACS technics.

关 键 词：循环肿瘤细胞 细胞角蛋白 流式细胞技术 

分 类 号：R730.4[医药卫生—肿瘤]