近致死量照射CNE1后存活克隆生物学性状初步分析  

Analysis of biological characteristics of survival clones from CNE1 cells irradiated with approximately-lethal dose

作　　者：冯雪萍[1] 刘伟 肖志强[1] 黄生鹏[3] 李建玲[4] 易红[4] 张鹏飞[1] 段朝军[1] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,湖南长沙410078 [2]长沙市163医院急诊科,湖南长沙410003 [3]中南大学湘雅医院放疗中心,湖南长沙410078 [4]中南大学湘雅医院医学实验研究中心,湖南长沙410078

出　　处：《中国耳鼻咽喉颅底外科杂志》2007年第4期245-249,共5页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：国家重点基础研究发展规划项目(973)(2001CB510207);国家自然科学基金资助项目(30000028;30670990);教育部跨世纪优秀人才培养计划基金(教计函[2002]48);湖南省科技厅一般项目(03SSY4019;01JZY2017);湖南省自然科学基金资助项目(03JJY4021);湖南省卫生厅重点科研项目(Z02-04)

摘　　要：目的放疗后残存癌细胞重新克隆是鼻咽癌放疗复发的主要原因,本实验旨在分析照射残存癌细胞的生物学特征。方法MTT法测定6MV-X射线照射人鼻咽高分化鳞癌细胞株CNE1的近致死剂量,用近致死量照射获得残存癌细胞存活克隆,然后用流式细胞仪检测存活克隆与亲本细胞株的细胞周期分布,应用二维电泳和质谱技术分析两者的蛋白质表达改变。结果用15Gy近致死量照射CNE1得到了照射存活克隆,并命名为CNE1-15GyR。CNE1-15GyR与亲本细胞株相比:G1/G0期比例增高而G2/M期比例降低;有98个蛋白质表达差异点,其中76个蛋白质斑点上调,22个蛋白质斑点下调;对其中上调的20个蛋白质斑点进行了质谱鉴定,发现细胞间叶来源的中间丝蛋白Vimentin在存活克隆CNE1-15GyR中表达增高。结论照射筛选的存活克隆CNE1-15GyR与亲本细胞株CNE1相比生物学性状发生了改变,细胞间叶来源的中间丝波形蛋白Vimentin在CNE1-15GyR克隆中表达增高,可能与G/G期比例增高有关。Objective To analyze the biological characteristics of the survival clones from nasopharyngeal carcinoma（ NPC） cell line CNE1 cells irradiated with approximately-lethal dose. Methods The approximately-lethal dose of 6MV-X-ray for CNE1 was determined by MTT assay, and the cells were exposed to this dose. The survival and the parent CNE1 cells were tested for the cell cycle distribution by the flow cytometry, and the changes of their proteins were analyzed by two-dimensional gel electrophoresis and MALDI-TOF-MS. Results The CNEl-15GyR survival cell clones were attained after CNE1 cells were irradiated with 6MV-X-ray of 15 Gy. Compared with the parent CNE1, higher G1/G0 and lower G2/M and 98 different protein spots （up-regulation at 76 spots and down- regulation in 22） were found in the CNE1-15GyR. The 20 up-regulated spots were identified by MALDI-TOF-MS. The expressions of intermediate filaments Vimentin from cell intermediate matrix were enhanced in CNE1-15GyR. Conclusion The biological characteristics of CNE1-15GyR have been changed compared with the parent cell lines. The enhanced expression of Vimentin in the CNE1-15GyR may be related to the higher ratio of G1/G0 in the cell cycle.

关 键 词：细胞株CNE1 X射线 存活克隆 二维电泳 丝波形蛋白(Vimentin) 鼻咽肿瘤 

分 类 号：R739.63[医药卫生—肿瘤] Q51[医药卫生—临床医学]