hTERTp在鼻咽癌中放射敏感性的研究  

Study of the radiosensitivity of hTERT promoterin nasopharyngeal carcinoma cell

作　　者：胡璟[1] 赵素萍[1] 唐瑶云[1] 

机构地区：[1]中南大学湘雅医院耳鼻咽喉科,湖南长沙410008

出　　处：《中国耳鼻咽喉颅底外科杂志》2009年第2期81-85,共5页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：湖南省自然科学基金资助;项目编号06JJ4050;高等学校学科点专项科研基金资助;项目编号20050533014;卫生厅项目资助;项目编号62006152

摘　　要：目的观察放射干预对人端粒酶逆转录酶基因(hTERT)启动子启动活性以及鼻咽癌靶向调控的影响。方法利用基因重组技术构建pGL-3-hTERTp质粒以及pcDNA3.1.hTERT.CD/UPRT质粒,脂质体介导重组质粒pGL-3-hTERTp转染鼻咽癌CNE-2细胞以及人成纤维皮肤细胞(HDF),并给予不同剂量放射干预,双荧光素酶活性分析其启动活性变化。pcDNA 3.1.hTERT.CD/UPRT质粒转染鼻咽癌CNE-2细胞;运用RT-PCR以及免疫印迹法检测不同放射剂量下自杀基因表达。结果接受放射干预后,hTERT启动子活性明显增强,给予2、6、1 0 Gy放射干预后,其启动活性分别增加了1 0倍、1 9倍和3 4倍,其差异有统计学意义(P<0.0 1)。HDF组无论放射与否,其测量值与pGL-3 Basic质粒无统计学差异(P>0.0 5)。pcDNA 3.1.hTERT.CD/UPRT质粒转染CNE-2细胞后,放射剂量增加至6、1 0 Gy检测到目的基因表达。结论放射干预可提高hTERT启动子在鼻咽癌CNE-2细胞中的启动活性,且对其肿瘤特异性无影响。以放疗为主的肿瘤治疗中,hTERT启动子可能为潜在靶向性基因治疗的备选因子。Objective To explore the influence of radiation on the promotive effect of hTERT promoter （hTERTp） and its target regulation in nasopharyngeal carcinoma cell line （CNE-2 cells ）. Methods Plasmids pGL-3-hTERTp and pcDNA3. 1. hTERT. CD/UPRT were reconstructed. Plasmid pGL-3-hTERTp was transfected into CNE-2 cells and HDF cells separately. After these ceils were treated with radiation of different doses , their promotive effect of hTERTp were detected with the Dual-Luciferase system. Plasmid pcDNA3.1, hTERT. CD/UPRT was transfected into CNE-2 cells. The transfected cells were given radiation in different dosage and then the expression of suicide gene was analyzed with the RT- PCR and Western blot. Results The Dual-Luciferase analysis showed that the promotive activities of hTERT promoter in pGL-3-hTERTp- transfected CNE-2 cells increased by 10, 19 , and 34 times after radiation in the doses of 2 Gy, 6 Gy, and 10 Gy, respectively, with significant statistical differences （ P 〈 0. 01 ）. As for the pGL-3-hTERTp- transfected HDF cells , no matter treated with radiation or not , the differences of the activities of hTERT promoter were statistically insignificant when compared with that of the reconstrueted plasmid （ P 〉 0.05 ） . The RT-PCR and Western blot revealed that the expression of the suicide gene could be detected in pcDNA3. 1. hTERT. CD/UPRT- transfected CNE- 2 cells when the radiation dose was up to 6 Gy. Conclusion Radiation can elevate the promotive activity of hTERTp in CNE-2 cells but does not change the specificity of the tumor, hTERTp is a potential promoter of target gene- therapy and is the main remedy for tumors with radiotherapy.

关 键 词：HTERT启动子 鼻咽癌 基因-放射治疗 

分 类 号：R739.6[医药卫生—肿瘤]