HRE1.Egr-1.yCDglyTK融合自杀基因前药系统对鼻咽癌放射增敏作用的体外实验  

Killing effect improved by fusion gene HRE1.Egr-1.yCDglyTK on gene-radio therapy of nasopharyngeal cancer in vitro

作　　者：钟宇[1] 唐瑶云[1] 谢常宁[1] 赵素萍[1] 

机构地区：[1]中南大学湘雅医院耳鼻喉科,长沙410008

出　　处：《中南大学学报（医学版）》2008年第2期110-114,共5页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(30300414)~~

摘　　要：目的:构建缺氧/辐射双敏感性启动子HRE1.Egr-1,观察其在缺氧或放射诱导下调控yCDglyTK基因在鼻咽癌HNE-1细胞表达及yCDglyTK/5-FC对鼻咽癌HNE-1细胞杀伤效应,为探索新的鼻咽癌基因-放射治疗奠定实验基础。方法:利用基因重组技术构建嵌合启动子HRE1.Egr-1调控yCDg-lyTK基因表达的质粒载体;脂质体介导重组质粒转染鼻咽癌HNE-1细胞,建立稳定表达yCDglyTK基因的鼻咽癌HNE-1转染细胞;免疫印迹法检测在常氧、放射、乏氧、乏氧加放射等不同条件下HNE1细胞中yCDglyTK表达的强度;MTT法检测加入5-FC后对上述不同条件下稳定转染细胞的杀伤效应。结果:各组蛋白表达均有差别(P<0.01),以乏氧/放射处理最为明显;5-FC对各种条件干预下的细胞均有杀伤效应,其中以乏氧/放射处理最为明显(P<0.01)。结论:HRE1.Egr-1嵌合启动子具有放射和乏氧诱导的双重特性,可以显著增强yCDglyTK/5-FC系统在乏氧和放射干预下对鼻咽癌HNE-1细胞的杀伤效应,为鼻咽癌的基因-放射治疗开辟了新思路。Objective To construct hypoxia/radiation inducible promotor HRE1. Egr-1 , and to observe its promotive effect on the expression of yCDglyTK gene in nasopharyngeal cancer HNE-1 cells and the anti-tumor effect of yCDglyTK and to lay an experimental foundation for further exploration of new gene-radio therapy of nasopharyngeal cancer. Methods pcDNA3.1 （ - ） HRE1. Egr-1. yCDglyTK was constructed by gene recombination technique. Stable yCDglyTK-expressing HNE-1 cells were generated by transfecting the recombinant plasmid into the target cells with liposome. The expression of yCDglyTK was detected by Western blot in 4 groups： a normoxia group, a radiation group, a hypoxia group, and a hypoxia and radiation group. The killing effect of 5-FC in different circumstances was determined by MTT. Results The expression of yCDglyTK/5-FC gene in all the groups was significantly different （ P 〈 0.01 ） , especially in the hypoxia and radiation group. The kill- ing effect of 5-FC on HNE 1 cells varied under different conditions, especially in the hypoxia and radiation group. Conclusion Hypoxia and radiation can induce the activity of fusion promoter HRE 1. Egr-1, and obviously promote the anti-tumor effect of yCDglyTK/5-FC system, suggesting that yCDglyTK may be a candidate suicide gene for gene-radio therapy of NPC.

关 键 词：鼻咽癌 缺氧反应元件 乏氧 基因-放射治疗 

分 类 号：R739.63[医药卫生—肿瘤]