反义角蛋白13慢病毒质粒对鼻咽癌细胞放射敏感性的影响  

Effect of antisense KRT13 lentivirus plasmid on the radiosensitivity of nasopharyngeal carcinoma cells

作　　者：刘曼 余远林[3] 高雪[1] 施明[1] 王欢[1] 余宏[1] 

机构地区：[1]云南省第二人民医院耳鼻咽喉科,云南昆明650000 [2]昆明医科大学,云南昆明650000 [3]华中科技大学同济医学院第二临床学院,湖北武汉430000

出　　处：《中国耳鼻咽喉颅底外科杂志》2014年第2期101-106,共6页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：云南省科技厅昆明医科大联合专项资金资助(2012FB084)

摘　　要：目的探讨反义角蛋白13(Keratin-13,KRT13)慢病毒质粒对鼻咽癌HNE-1细胞放射敏感性的影响。方法构建稳定表达反义KRT13基因的慢病毒质粒,并通过G418筛选出稳定转染该质粒的鼻咽癌HNE-1细胞。按数字随机法将细胞分为control(正常HNE-1细胞)组、lentivirus(转染慢病毒空载体)组和anti-KRT13(转染慢病毒质粒组)。并分别检测各组细胞中KPT13含量。同时采用流式细胞术检测细胞周期和凋亡的变化。在0、1、2、4、6、8 Gy 6个放射剂量点下,采用克隆形成实验分析转染反义KRT13基因对细胞的存活影响,并分别用单击多靶模型和线性二次模型拟合出细胞的剂量存活曲线,对比放射生物学参数D0、Dq、N值、α、β和SF2值,评价细胞放射敏感性的变化。结果转染慢病毒质粒组的HNE-1细胞D0、Dq、N和SF2值较其他两组均增加,α/β值减小,且差异具有统计学意义(P均<0.05);反义KRT13可以阻滞细胞于G2/M期,同时抑制细胞凋亡。结论反义KRT13慢病毒质粒可降低鼻咽癌HNE-1的放疗敏感性。Objective To explore the effect of antisense KRT13 lentivirus plasmid on the radiosensitivity of nasopharyngeal carcinoma cells （HNE-1）. Methods The lentivirus package system carrying antisense RNA of KRT13 was constructed, and stable expressed antisense KRT13 HNE-1 cell line was screened by G418. The cells were randomly divided into three groups, control, lentivirus and anti-KRT13 group, and to detect the content of KRT13 in each group. Cells were exposed to radiation of various dosage（0, 1 , 2, 4, 6 and 8 Gy） , and then the clonogenie survival and curve fitting were used for calculating the radiobiological parameters and the sensitization enhancement ratio after radiation. The apoptosis and cell cycle changes were analyzed by flow cytometry. Results Compared with the control cells and negative ceils, anti-KRT13 cells showed obviously high values of DO, Dq, N, and SF2 but significantly decreased cd[3value and the differences were statistically significant （P 〈 0. 05 ）. Anti- KRT 13 could block HNE-1 cells in G2/M phase and inhibit the apoptosis. Conclusion Antisense KRT 13 lentivirus plasmid can reduce the radiosensitivity of HNE-1 cells in vitro.

关 键 词：角蛋白13 反义慢病毒 鼻咽癌HNE-1细胞 放射敏感性 

分 类 号：R739.63[医药卫生—肿瘤] Q786[医药卫生—临床医学]