人喉癌细胞异质性亚群的克隆分离及性质初探  

Preliminary study on subpopulation cell isolation and heterogeneity characterization of human laryngeal carcinoma

作　　者：于丹[1] 金春顺[1] 赵黎明[2] 陈鸥[1] 

机构地区：[1]吉林大学第二医院耳鼻咽喉科,吉林长春130041 [2]北京市耳鼻咽喉科研究所,首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,耳鼻咽喉头颈科学教育部重点实验室(首都医科大学),北京100005

出　　处：《中国耳鼻咽喉头颈外科》2009年第1期17-20,共4页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：吉林省科技厅重大社会发展项目(20060416);吉林省科技厅应用基础项目(200705194)联合资助

摘　　要：目的通过原代培养人喉癌细胞、克隆分离异质性细胞亚群,初步探讨喉癌异质性机制。方法原代培养人喉癌细胞,采用有限稀释法单细胞体外培养喉癌细胞,比较克隆分离得到的异质性亚群在功能状态、细胞周期、形态、在无血清干细胞培养基中生长情况及裸鼠异体移植成瘤能力等方面的差异。结果分离得到Ⅰ型和Ⅱ型两个具有明显形态差异的细胞亚群。Ⅰ型细胞呈多角形,突起多,裸鼠移植可成瘤(4/5);而Ⅱ型细胞呈长梭形,突起较少,裸鼠异体移植不成瘤(0/5)。Ⅰ型与Ⅱ型细胞相比,RNA含量低,体外增殖能力强,能在无血清干细胞培养基中聚集呈肿瘤细胞球状生长;Ⅰ型细胞处于细胞周期G0/G1的比例明显高于Ⅱ型细胞。对Ⅰ型细胞亚克隆培养,产生的子代克隆集落出现了异质性的细胞,而Ⅱ型细胞亚克隆培养未能产生子代细胞。结论原发性喉癌细胞中存在着异质性的肿瘤细胞亚群,Ⅰ型细胞具备肿瘤干细胞的特性。OBJECTIVE To isolate and characterize the heterogeneous subpopulation cells from human laryngeal carcinoma, and to explore the mechanism of heterogeneity in laryngeal carcinoma. METHODS Laryngeal carcinoma cells were cultured in vitro by primary tissue culture technique. Heterogeneous subpopulations were isolated by limiting dilution and were identified their functional status （mRNA） , cell cycles （G0/G1） , morphology and growth condition in serum-free RPMI1640 medium. Each subpopulation was implanted in nude mice subcutaneously to measure the tumor forming ability. RESULTS Two different subpopulations of type I and type II laryngeal carcinoma cells were derived from primary tumor tissues. Type I cells were multiangular in shape with protuberances and could produce new tumor （4/5） . However, type II cells showed a spindle feature and could not form new tumor when implanted in nude mice （0/5） . Compared with type II cells, type I cells contained much less RNA and more G0/G1 cells, demonstrated increased proliferation capacity and can grow in serum-free RPMI1640 medium like tumor spheres. Furthermore, type I cells can create heterogeneous subclone offspring, but type II cells can not generate offspring cells. CONCLUSION Laryngeal carcinoma cells are heterogeneous with respect to characteristics on morphology, proliferation, functional and growth status. Type I cells possess some characteristics of tumor stem cells.

关 键 词：喉肿瘤 遗传异质性 肿瘤干细胞 

分 类 号：R739.65[医药卫生—肿瘤]