应用免疫激活磁珠分选技术CD45去除方法富集——免疫细胞化学联合苏木素-伊红染色检测肝癌患者循环肿瘤细胞  

Combined use of immunomagnetic activated cell sorting technique enrichment and immunocytochemistry with hematoxylin and eosin staining for identification of circulating tumor cells in peripheral blood mononuclcar cells of hepatocellular carcinoma patients

作　　者：郭立民[1] 鲁岩[1] 彭吉润[2] 蒋力[1] 

机构地区：[1]首都医科大学附属北京地坛医院外科, 100015 [2] 首都医科大学附属北京世纪坛医院外科

出　　处：《中华实验外科杂志》2014年第9期2019-2021,共3页Chinese Journal of Experimental Surgery

基　　金：首都医科大学附属北京地坛医院院内青年科研基金资助项目（QN2011-05）

摘　　要：目的 探讨免疫激活磁珠分选（MACS） CD45去除方法富集后,以细胞角蛋白（CK）为标记联合苏木素-伊红（HE）染色检测肝癌患者外周血肿瘤细胞（CTC）的价值.方法 应用逆转录-聚合酶链反应（RT-PCR）检测18种上皮肿瘤细胞株的CK（CK8、CK18、CK19）表达;采集健康志愿者、肝癌患者外周血,以MACS技术CD45去除方法对外周血单个核细胞（PBMC）进行富集,以CK为标记,采用免疫组织化学染色联合HE染色检测肿瘤细胞.结果 在18种上皮性肿瘤细胞株中CK8、CK18、CK19表达率分别为72.22％、83.33％和66.67％;9种肝癌细胞株中CK阳性表达占绝对多数;在MACS技术CD45去除富集肿瘤细胞基础上,应用免疫细胞化学和HE染色结果显示,肝癌患者外周血存在完整肿瘤细胞,检出率为63.15％ （12/19）,健康志愿者未检测到肿瘤细胞.结论 CK可作为检测外周血肝癌细胞标志物;MACS技术CD45去除方法富集后,CK免疫细胞化学和HE染色同步检测方法有助于检测肝癌患者外周血肿瘤细胞.Objective To estimate the applied value of magnetic activated cell sorting （MACS） techniques with CD45 depletion and immunocytochemistry in combination with hematoxylin and eosin （HE） staining in identifying circulating tumor cells （CTCs） in peripheral blood mononuclear cells （PBMC） of hepatocellular carcinoma （HCC） patients.Methods The expression of CK （CK8,CK18,and CK19） was detected in 18 epithelia-derived tumor cell lines including 9 human hepatocellular carcinoma cell lines.The peripheral blood of HCC patients and healthy volunteers was collected for determination of CTCs in PBMC from HCC patients using MACS techniques with CD45 depletion and immunocytochemstry in combination with HE staining.Results The expression rate of CK8,CK18 and CK19 in the selected CTCs was 72.22％,83.33％ and 66.67％ respectively.CKs were detected in most of the 9 hepatocelluar carcinoma cell lines.We found intact CTCs in PBMC from HCC patients using HE staining and immunocytochemistry after PBMC enrichment by MACS techniques with CD45 ＋ depletion.The sensitivity of this method was up to 63.15％,and no CTCs were detected in PBMC from 20 healthy controls.Conclusion CKs could be a tumor marker for detection of CTCs in HCC patients.The method of HE staining and immunocytochemistry after PBMC enrichment by MACS technique with CD45 ＋ depletion has potentials in detection of circulating HCC cells.

关 键 词：循环肿瘤细胞 细胞角蛋白 免疫激活磁珠分选 苏木素-伊红染色 

分 类 号：R735.7[医药卫生—肿瘤]