人喉癌Hep-2细胞株CD133＋肿瘤干细胞生物学特性研究  

Investigation on biological characteristics of CD133＋ cancer stem cells in human laryngeal carcinoma Hep-2 cell line

作　　者：卫旭东[1] 何健[1] 高江霞[1] 王晶宇[2] 马斌娟[1] 陈静[1] 

机构地区：[1]甘肃省人民医院耳鼻咽喉头颈外科,兰州730000 [2]兰州大学基础医学院病理生理学研究所

出　　处：《中华耳鼻咽喉头颈外科杂志》2013年第7期578-583,共6页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的 研究喉癌Hep-2细胞株中CD133＋肿瘤细胞的干细胞特性及其生物学特性.方法 流式细胞仪分选Hep-2细胞株中CD133＋肿瘤细胞,Transwell法检测CD133＋肿瘤细胞侵袭能力及三代克隆形成能力；CD133＋肿瘤细胞与紫杉醇共培养,10 Gy的直线加速器照射CD133＋肿瘤细胞,四甲基偶氮唑蓝法计算其相对存活率和生长抑制率,观察其放化疗抵抗性.结果 CD133＋细胞比例为3.1％±0.2％,流式细胞仪分选纯度可达90.2％±5.5％,分选后的CD133＋细胞贴壁,呈团块状、克隆球状生长.增殖速度明显较CD133-细胞快；Transwell实验中,相同视野CD133＋细胞穿膜细胞数（526±39）个/每视野,而CD133-细胞为（220±20）个/每视野,二者差异有统计学意义（t=22.08,P＜0.01）；CD133＋细胞三代克隆形成率分别为30.0％±4.7％、32.2％±3.6％、32.7％±3.4％,CD133-细胞三代克隆形成率分别为15.2％±2.2％、12.0％±2.5％、13.8％±3.3％,同代比较差异有统计学意义（t值分别为8.99、14.66、12.69,P值均＜0.01）.在紫杉醇共培养体系中,CD133＋细胞24 h、48 h、72 h的相对存活率分别为90.1％±5.9％、85.1％±7.1％、70.3％±6.4％,均高于同时期对照组CD133-细胞（t值分别为5.24、8.18、8.14,P值均＜0.01）；放射线照射后,CD133＋细胞生长抑制率30.0％±7.1％,显著低于CD133-细胞的55.0％±6.3％（t=8.30,P＜0.01）.结论 CD133＋ Hep-2细胞所占比例较小,具有强的增殖、侵袭能力及克隆形成能力,对放化疗有抵抗性,具有干细胞特性.靶向CD133＋肿瘤细胞,有望有效杀伤喉癌干细胞.Objective To investigate the biology characters of CD133 ＋ cancer stem cells from Hep-2 cell line.Methods The Flow cytometry was applied to purify CD133 ＋ cells from Hep-2 cell line.The sorted CD133 ＋ cells were cultured in RPMII640.The ability of migration,invasion and clonality of CD133 ＋ cells were performed to characterize the properties of the cells.CD133 ＋ cells and CD133-control cells were treated with paclitaxel and exposed respectively to X-rays emitted by linear accelerator with a dose of 10 Gy.The surviving rates and growth inhibition ratio of cells in two groups were detected with MTT assay to observe the resistance to irradiation and chemotherapy in CD133 ＋ cells.Results The percentage of CD133＋ cells in the unsorted and the sorted cells were 3.1% ±0.2% and 90.2% ±5.5%,respectively.CD133＋ cells showed the higher proliferation and colony ability than CD133-cells.The numbers of CD133 ＋ and CD133-cells that passed the membrane of Transwell chamber were 526 ± 39 and 220 ± 20 respectively（t =22.08,P 〈 0.001）.The colony forming units of three passages were 30.0% ± 4.7%,32.2% ±3.6%,32.7% ±3.4% in CD133＋ cells and 15.2% ±2.2%,12.0% ±2.5%,13.8% ±3.3%in CD133-cells.There were statistic difference between two groups（t =8.99,t =14.66,t =12.69,P 〈0.01）.At 24,48,and 72 hours of treatment with paclitaxel,the cell surviving rates of CD133 ＋ cells were 90.1% ±5.9%,85.1% ±7.1% and 70.3% ±6.4% and lower than those of CD133-cells,respectively （t =5.24,t =8.18,t =8.14,P 〈 0.01）.After radiotherapy,growth inhibition ratio of CD133 ＋ and CD133-cells were 30.0% ±7.1% and 55.0% ±6.3%（t =8.30,P〈0.01）.Conclusions CD133＋ cells exist in a small proportion in Hep-2 cell line and they show the properties of cancer stem cells,with the resistance to irradiation and chemotherapy.

关 键 词：喉肿瘤 细胞系 肿瘤 肿瘤干细胞 肿瘤浸润 克隆细胞 辐射耐受性 抗药性 肿瘤 

分 类 号：R739.65[医药卫生—肿瘤]