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 CD133+ SP及CD133 - SP细胞在喉癌Hep-2细胞系中各占(0.30±0.12)％及(17.52±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((-x)±s,下同)为(0.36±0.15)g,而CD133 - SP细胞所成瘤体重量平均为(0.08 ±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.

abstractsObjective 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 right 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 helped to purify CSCs further from laryngeal cancer cell line.