| 廖正凯,周云峰,谢丛华,等.γ射线增强喉癌细胞hTERTp启动子活性研究[J].中华放射医学与防护杂志,2008,28(2):104-107.LIAO Zheng-kai,ZHOU Yun-feng,XIE Cong-hua,et al.Enhancement of hTERT promoter activity by γ-rays in laryngeal squamous carcinomas cells in vitro[J].Chin J Radiol Med Prot,2008,28(2):104-107 |
| γ射线增强喉癌细胞hTERTp启动子活性研究 |
| Enhancement of hTERT promoter activity by γ-rays in laryngeal squamous carcinomas cells in vitro |
| 投稿时间：2007-01-13 |
| DOI： |
| 中文关键词: γ射线 放射耐受 hTERT启动子 喉癌 HRP/IAA |
| 英文关键词:Gamma rays Radiation tolerance hTERT promoter Laryngeal squamous cell carcinoma HRP/IAA |
| 基金项目:国家自然科学基金资助项目(30672438);湖北省自
然科学基金创新群体资助项目(2006ABC009) |
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| 中文摘要: |
| 目的 探讨γ射线对喉癌细胞中端粒酶逆转录酶基因启动子(hTERTp)活性的影响,以及通过射线增强hTERTp下游基因表达的可行性。方法 将含hTERTp的质粒转染细胞,采用报告基因评价启动子活性,通过RT-PCR和酶活性检测观察射线作用下hTERTp调控辣根过氧化物酶(HRP)的表达,克隆形成实验评价射线对phTERTp-HRP/IAA杀伤和放射增敏作用的影响。结果 在Hep-2细胞中,6 Gyγ射线照射后hTERTp活性是0 Gy照射后的2.96倍,在Hep-2R细胞中是1.60倍。质粒phTERTp-HRP转染Hep-2和Hep-2R细胞后,6 Gy照射组的HRP mRNA分别增高2.1和1.1倍,酶活性分别增高2.54和1.23倍。6 Gy联合吲哚乙酸(IAA)组Hep-2细胞的存活分数为1.3%,Hep-2R细胞的存活分数为3.5%,比对照组明显降低(F=234.280和F=357.148,P值均<0.01)。6Gy联合IAA组与IAA组相比,放射增敏比SER<sub>SF2分别为1.52(Hep-2)和1.68(Hep-2R),存活曲线的参数α分别为0.416、0.099(Hep-2)和0.356、0.090(Hep-2R)。结论 γ射线可以增强不同放射敏感性喉癌细胞hTERTp活性,增强下游基因表达。射线联合phTERTp-HRP/IAA对喉癌细胞具有更加明显的杀伤和放射增敏作用。 |
| 英文摘要: |
| Objective To investigate the effect of γ-rays on hTERT promoter activity in laryngeal squamous carcinomas cell lines, and to evaluate the efficiency of hTERTp mediated gene therapy combined with γ-rays.Methods hTERTp activity was determined by luciferase assay. Plasmids phTERTp-HRP were transfected into cells, HRP expression levels were determined by RT-PCR and enzyme activity assay. The cytotoxicity and radiosensitivity of phTERTp-HRP/IAA were determined by clonogenic assay.Results After 6 Gy irradiation, a 2.96-fold increase of hTERTp activity in Hep-2 cells and a 1.60-fold increase in Hep-2R cells were found. Hep-2 and Hep-2R cells transfected with phTERTp-HRP, HRP mRNA expression levels were 2.1-fold and 1.1-fold increased, while enzyme activity of HRP were increased by 2.54-fold and 1.23-fold, respectively after 6 Gy irradiation. Combination of 6 Gy induction and IAA incubation obviously decrease the surviving fraction of Hep-2 and Hep-2R cells (P<0.01), and clearly radiosensitize the cells without induction (SER<sub>SF2=1.52 in Hep-2; SER<sub>SF2=1.68 in Hep-2R), the parameterα with or without 6Gy induction before IAA incubation were 0.416, 0.099(Hep-2)and 0.356, 0.090(Hep-2R),respectively.Conclusions γ-rays can enhance hTERT promoter activity and improve the expression of downstream gene in different radiosensitive laryngeal squamous carcinomas cells. Combination of radiation and hTERTp-HRP/IAA construct results in significant cytotoxicity and radiosensitization. |
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