缺氧/辐射双敏感性启动子调控CD/UPRT融合自杀基因在Hep-2细胞表达的实验研究  

Expression of CD/UPRT fusion suicide gene controlled by a hypoxia/radiation dural sensitive promoter in Hep-2 cells

作　　者：唐瑶云[1] 邓小云 徐婧[1] 张俊毅[1] 刘庭[3] 刘建平[4] 肖健云[1] 赵素萍[1] 

机构地区：[1]中南大学湘雅医院耳鼻咽喉科,湖南长沙410008 [2]湖南省经济发展改革委员会,湖南长沙410004 [3]中南大学湘雅医院消化内科,湖南长沙410008 [4]中南大学现代分析测试中心,湖南长沙410078

出　　处：《中国耳鼻咽喉颅底外科杂志》2006年第5期321-325,共5页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：国家自然科学基金(编号:30300414)

摘　　要：目的本研究拟构建缺氧/辐射双敏感性启动子,观察其在缺氧或放射诱导下调控CD/UPRT基因在喉癌Hep-2细胞的表达水平,为探索新的喉癌基因-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CD/UPRT基因表达的质粒表达载体;脂质体介导重组质粒转染喉癌Hep-2细胞,分为对照组、放射组(2、4、6、8 Gy)、缺氧组(1%、2%3、%O2浓度)及放射加缺氧组(8 Gy+3%O2),Western-blotting法检测转染细胞中CD/UPRT表达。结果对照组细胞仅可检测到CD/UPRT的低水平表达,放射组蛋白表达的相对灰度值分别为1.3、1.6、1.8、2.9,缺氧组CD/UPRT基因表达相对灰度值分别为1.5、2.0、2.2,放射合并缺氧组CD/UPRT表达显著高于放射组或者缺氧组(P<0.01)。结论HRE1.Egr-1启动子具有辐射/缺氧双重敏感性,并使放射干预后的Hep-2细胞CD/UPRT表达水平在缺氧下得到显著增强,为喉癌的基因-放射治疗开辟了新思路。Objective To construct a hypoxia/radiation dural sensitive promoter and observe the fusion suicide gene CD/UPRT expression driven by the promoter in transfected Hep-2 cells for exploring a new gene-radiotherapy of laryngeal cancer. Methods Plasmid containing CD/UPRT gene and its regulation promoter HRE1. Egr-1 was generated by recombinant gene technic and transfected into Hep-2 cells using lipofectamine. Such transferred cells were intervened with different conditions （control group： no interference; radiation group：exposed to 2, 4, 6, 8 Cry X-ray respectively; hypoxia group： treated with 1%, 2%, 3% O2 respectively; radiation/hypoxia group： in the presence of 8 Gy X-ray and 3% O2 ）, then CD/UPRT expressions were analyzed using Western blot. Results Low level of CD/UPRT protein was detected in control group, whereas the relative grey values of expression protein CD/ UPRT were 1.3, 1.6, 1.8, 2.9 in radiation group and 1.5, 2. 0, 2. 2 in hypoxia group respectively. Compared with radiation group or hypoxia group, the expression of CD/UPRT protein in radiation/hypoxia group increased significantly （P 〈0. 01）. Conclusion HRE1. Egr-1 promoter possesses hypoxia and radiation dural sensitive characters and can extinctly enhance the CD/UPRT expression in Hep-2 cells exposed to Xray under hypoxia. This special property makes of HRE1. Egr-1 promoter a novel candidate of gene-radiotherapy strategies for laryngeal cancer.

关 键 词：喉肿瘤/放射疗法 基因-放射治疗 缺氧反应元件 

分 类 号：R739.65[医药卫生—肿瘤]