人结直肠癌细胞株来源的HCT116肿瘤细胞球的转移相关特性  

Metastasis-related characteristics of tumor spheres derived from human colorectal cancer cell line HCT116

作　　者：贾健锋[1] 程勇[1] 武乃金[1] 任林飞[1] 徐维[1] 刘弈武[1] 杨小丁[1] 

机构地区：[1]重庆医科大学附属第一医院胃肠外科,重庆400016

出　　处：《肿瘤》2011年第11期982-986,共5页Tumor

基　　金：重庆市自然科学基金资助项目(编号:2008BB5230)

摘　　要：目的:无血清悬浮培养生成HCT116肿瘤细胞球,检测球中CD133表面标志分子的细胞定位,以及CD133+细胞所占的比例,并探索肿瘤细胞球发生上皮-间质转化与其转移能力之间的可能联系。方法:HCT116细胞在添加了细胞生长因子的无血清培养液(serum-freemedium,SFM)中悬浮培养生成细胞球,应用激光共聚焦和流式细胞仪检测CD133表面标志分子的细胞定位及细胞球中CD133+细胞的含量。Transwell小室实验、免疫荧光及反转录PCR(reverse transcription-PCR,RT-PCR)法分别检测细胞球的体外转移能力以及上皮-间质转化关键分子的表达情况。结果:HCT116细胞球中CD133表面标志分子定位于各个球层面的细胞膜,并且CD133+细胞比例显著增多。细胞球高表达间质化分子N-cadherin及Vimentin,而上皮化分子E-cadherin表达水平下降,并具有更强的转移能力。结论:HCT116细胞球中富含CD133+细胞,CD133表面标志分子定位于细胞膜,细胞球可能通过上皮-间质转化而获得更强的转移能力。Objective：To generate tumor spheres from colorectal cancer cell line HCT116 in serum-free medium（SFM） and to observe the location of the cancer stem cell surface marker CD133 as well as the ratio of CD133＋ cells so as to investigate the possible relationship between the epithelial-mesenchymal transition and metastatic ability of tumor spheres.Methods：Human HCT116 colorectal cancer cells were cultured in SFM supplemented with cell growth factors.The location of cancer stem cell marker CD133 in tumor spheres was observed by confocal laser scanning microscopy,and the content of CD133＋ cells in tumor spheres was detected by flow cytometry（FCM）.Transwell chamber assay,immunofluorescence and reverse transcription polymerase chain reaction（RT-PCR） were applied to examine the metastatic ability of tumor spheres in vitro and the expressions of epithelial-mesenchymal-related key markers.Results：The surface marker CD133 was located on cell membrane,and the content of CD133＋ cells was significantly increased in HCT116 tumor spheres.The expressions of mesenchymal markers N-cadherin and Vimentin were highly elevated,while the expression of epithelial marker E-cadherin was reduced.The metastatic ability of cell sphere was higher than that of monolayer cells.Conclusion：HCT116 tumor spheres are enriched with CD133＋ cells.The surface marker CD133 is located on the cell membrane.The cell spheres may get higher metastatic ability through epithelial-mesenchymal transition.

关 键 词：结直肠肿瘤 肿瘤干细胞 球形体 细胞 抗原 CD133 上皮-间质转化 

分 类 号：R737.9[医药卫生—肿瘤]