喉癌CD133^＋侧群细胞的分离及体内致瘤性分析  

Isolation and in vivo tumorigenicity assay of CD133 ＋ side population cells from laryngeal cancer cellline

作　　者：吴春萍[1] 周梁[1] 谢明[1] 陶磊[1] 张明[1] 田洁[2] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻咽喉头颈外科,上海200031 [2]复旦大学附属眼耳鼻喉科医院中心实验室,上海200031

出　　处：《中华耳鼻咽喉头颈外科杂志》2012年第3期223-227,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：上海市科委自然科学基金（08ZR1402500、08JC1404000）

摘　　要：目的探索能够有效富集喉癌肿瘤干细胞的分选方法。方法流式细胞仪辅助下联合应用CD133标记及侧群（side population，sP）细胞分选技术检测及分选喉癌Hep-2细胞系中的CD133^＋SP及CD133-SP细胞，将分选所得两亚群细胞以相同的数量级注入非肥胖糖尿病型／重症联合免疫缺陷（NOD／SCID）小鼠右侧腋窝皮下，比较两亚群细胞致瘤性差异，并对两亚群细胞进行细胞周期检测分析。结果CDl33^＋SP及CD133-SP细胞在喉癌Hep-2细胞系中各占（0．30±0．12）％及（17．524-1．59）％。CD133^＋SP在16只NOD／SCID小鼠中的15只腋窝皮下成瘤，CD133-SP细胞仅在16只NOD／SCID小鼠中的7只腋窝皮下成瘤，差异有统计学意义（Fisher精切概率法，P〈0．05）。CD133^＋SP细胞所成瘤体的重量平均（-xs±，下同）为（0．364-0．15）g，而CD133-SP细胞所成瘤体重量平均为（0．084-0．04）g，差异有统计学意义（t=4．64，P〈0．01）。细胞周期检测发现，CD133^＋SP及CD133-SP细胞具有相似的细胞周期分布。结论CD133^＋SP比CD133-SP具备更强的免疫缺陷小鼠体内致瘤能力；SP细胞分选法与CD133标记相结合是一种更有效的富集喉癌肿瘤干细胞的分选方法。Objective To investigate a valuable strategy for further purifying cancer stem cells （CSCs） from laryngeal cancer cell line. Methods CD133 ＋ side population （SP） and CD133- SP cells were detected and isolated from laryngeal cancer Hep-2 cell line with SP discrimination and CD133 surface marker, assisted by fluorescence activated cell sorting technology. Freshly sorted CD133 ＋ SP and CD133 - SP cells were xenografted into the subcutaneous space of the fight axillary fossa of NOD/SCID mice and tumorigenic capacity of the cells from two subgroups were examinated. Cell cycle distributions of the two cell populations were detected. Results CD133 ＋ SP and CD133 - SP cells accounted for （0. 30 ± 0. 12）% and （ 17.52 ± 1.59 ）% in Hep-2 cell line, respectively. CD133 ＋ SP cells formed tumor nodules in 15 of 16 mice and CD133 - SP cells in 7 of 16 mice （ Fisher＇s exact test,P 〈0. 05）. The mean weight of CD133 ＋ SP tumor nodules was （0. 36 ±0. 15）g and that of CD133 - SP tumor nodules was （0. 08 ±0. 04） g. The difference was significant （ t = 4. 64, P 〈 0. 01 ）. Cell cycle analysis revealed similar cycle distributions between the two subgroups. Conclusions CD133 ＋ SP cells harbored much more cancer stem-like tumorigenic potential in NOD/SCID mice than CD133- SP cells. The combination of SP discrimination and surface marker selection hehaed to ourifv CSCs further from laryngeal cancer cell line.

关 键 词：喉肿瘤 肿瘤干细胞 抗原 CD 侧群细胞 致癌性试验 

分 类 号：R730.264[医药卫生—肿瘤]