8号和20号染色体拷贝数改变辅助食管癌外周血循环肿瘤细胞鉴定  

An Analysis of Chromosome 8 and 20 Copy Number Changes to Assist Identification of Circulating Tumor Cells in Peripheral Blood of Esophageal Carcinoma

作　　者：任传利[1] 何平[1] 张金强[1] 乔媛媛[2] 王伟[2] 周兰萍[1] 文峰[2] 赵平[3] 赵晓航[1] 

机构地区：[1]中国医学科学院肿瘤研究所分子肿瘤学国家重点实验室,北京100021 [2]海军总医院,北京100037 [3]中国医学科学院肿瘤医院,北京100021

出　　处：《中国肿瘤》2008年第6期513-517,共5页China Cancer

基　　金：国家自然科学基金(30772507、30721001);“973”(2004CB518707);“863”(2006AA02Z19B、2006AA02A403、2006AA02Z341);博士学科点专项基金(20060023010)

摘　　要：[目的]建立基于细胞学鉴定食管鳞癌(ESCC)外周血循环肿瘤细胞(CTCs)的技术。[方法]通过偶联CD45抗体的免疫磁珠去除白细胞,富集食管癌外周血稀有细胞的"负性富集"策略,结合抗CK8/18/19免疫荧光染色(IF)和8/20号染色体荧光原位杂交(FISH)鉴定CTCs。[结果]食管癌外周血CK8/18/19+/CD45-的CTCs检出率为43.8%(7/16),其中80%伴有8/20号染色体非整倍体的改变(AC)。食管癌细胞WHCO1、EC0156和KYSE150的8号AC分别为77.3%、78.6%和75%;KYSE150的20号AC为77.9%。外周血一些胞浆胞膜不完整、具有异型性的细胞核伴有8/20号AC改变。[结论]食管癌8/20号AC可以作为判断食管癌外周血CTCs恶性表型的标准,结合CK8/18/19+/CD45标准,不但可以鉴定具有完整细胞结构的CTCs,还可以鉴定胞浆、胞膜不完整或裸核样的CTCs的恶性表型。[Purpose] To establish a cytology-based method to identify circulating tumor cells （CTCs） in peripheral blood of esophageal squamous cell carcinoma （ESCC）. [Methods] Magnetic beads labeled with anti-CD45 were used to deplete leukocytes and enrich rare tumor cells in peripheral blood of esophageal carcinoma, which was recognized as a negative selection of CTCs. And then the CTCs were identified by immunofluorence staining （IF） of CK8/18/19 and fuorescence in situ hybridization （FISH） of chromosome 8 and 20. [Results ] CTCs characterized CK8/18/19^＋/CD45^- were detected in 43.8% （7/16） peripheral blood samples, 80% of which were additionally found aneuploid chromosomes （AC） on chromosome 8 and 20. Some human esophageal carcinoma cell lines, such as WHCO1, EC0156 and KYSE150, were also found with a high frequency of AC on chromosome 8, 77.3%, 78.6% and 75.0%, respectively, 77.9% of AC on chromosome 20 in KYSE150 cells. Some epithelia cells with a fragmented plasma and membrane, or nude nuclei in peripheral blood of esophageal carcinoma were successfully identified as CTCs by detection of chromosome 8 and 20 copy number changes. [Conclusions] The AC on chromosome 8 and 20 can be as a c, riteria for identifying CTCs in peripheral blood of esophageal carcinoma. Integrate both the criteria of CK8/18/19^＋/CD45^- by IF and 8/20 aneuploid chromosomes by FISH, it could be properly identified CTCs not only the cells with intact structures but also the one with fragmented plasma membrane or nude nucleus.

关 键 词：食管鳞癌 循环肿瘤细胞 荧光原位杂交 

分 类 号：R73-3[医药卫生—肿瘤] R735.1[医药卫生—临床医学]