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.
出处
《中国肿瘤》
CAS
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)
关键词
食管鳞癌
循环肿瘤细胞
荧光原位杂交
esophageal squamous cell cancer
circulating tumor cells
fluorescence in situ hybridization
