人鼻咽癌放射抗拒性肿瘤干细胞的分离、筛选与鉴定  

Screening and identification of tumor stem cells in radioresistance human nasopharyngeal carcinoma

作　　者：王亚利[1] 王中卫[1] 任洪涛[1] 金迎迎[1] 李毅[1] 

机构地区：[1]西安交通大学医学部第二附属医院肿瘤科放疗中心,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2015年第5期650-654,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81141097)~~

摘　　要：目的分离、筛选、鉴定人鼻咽癌CNE-2及放射耐受CNE-2R的肿瘤干细胞。方法免疫磁珠技术分选CNE-2及CNE-2R中鼻咽癌CD133+干细胞,分别用无血清培养法、平板克隆法、裸鼠成瘤试验检测分选的CD133+细胞体外增殖能力、克隆形成能力及成瘤能力,HE染色和电镜超微结构观察形态学变化。结果 CNE-2及CNE-2R细胞中CD133+表达分别为(1.79±0.16)%和(2.63±0.72)%;CNE-2-CD133+细胞及CNE-2R-CD133+细胞形成肿瘤干细胞球及体外增殖能力远高于CD133-表达者;CNE-2R-CD133+细胞克隆形成能力较CNE-2-CD133+细胞明显升高,而且CNE-2R-CD133+细胞的克隆形态与CNE-2-CD133+细胞相比较明显增大;形态学发现CNE-2R-CD133+细胞核大,异型,核仁大,细胞器肥大。结论分离、筛选出的鼻咽CNE-2R-CD133+肿瘤干细胞能形成干细胞球,其增殖能力、裸鼠成瘤能力更强,能为进一步探讨鼻咽癌放射抗拒的机制提供研究基础。Objective To isolate,screen and identify human nasopharyngeal carcinoma CNE-2 and radioresistance CNE-2R cancer stem cells.Methods The CNE-2 and CNE-2R CD133＋ stem cells were sorted with flow cytometry and immunomagnetic beads.The CNE-2-CD133＋ and CNE-2R-CD133＋ stem cells were detected for the abilities of tumor sphere formation and proliferation in vitro.The clonogenic ability of CD133＋stem cell was detected by plate clone formation assay and tumorigenicity was detected by nude mice test in vivo.Morphological changes were observed with HE staining and electron microscopy.Results CD133＋ was expressed（1.79±0.16）% and（2.63±0.72）%in CNE-2 and CNE-2R cells,respectively.The clonogenic ability and tumor sphere formation ability of CNE-2R CD133＋ cells were much greater than those of CD133-cells.The ability of proliferation of CD133＋cell sorted in vitro,especially CNE-2R-CD133＋cells,was stronger than that of CD133-cells.The morphological results showed that the nucleus of CNE-2R-CD133＋ cells was larger and irregular with big nucleolus and organelles.Conclusion Nasopharyngeal CNE-2R-CD133＋ tumor stem cells isolated and screened can form tumor sphere.The abilities of proliferation and tumorigenesis are stronger.The identification of CNE-2R TSC provides foundation for further studies on the mechanism of radioresistance of nasopharyngeal carcinoma.

关 键 词：鼻咽癌 放射抗拒性 肿瘤干细胞 

分 类 号：R739.63[医药卫生—肿瘤]