LIAO Zheng-kai,ZHOU Yun-feng,XIE Cong-hua.Enhancement of hTERT promoter activity by γ-rays in laryngeal squamous carcinomas cells in vitro[J].Chinese Journal of Radiological Medicine and Protection,2008,28(2):104-107
Enhancement of hTERT promoter activity by γ-rays in laryngeal squamous carcinomas cells in vitro
Received:January 13, 2007  
DOI:
KeyWords:Gamma rays  Radiation tolerance  hTERT promoter  Laryngeal squamous cell carcinoma  HRP/IAA
FundProject: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)
Author NameAffiliationE-mail
LIAO Zheng-kai Department of Radio-Chemotherapy, Zhongnan Hospital and Cancer Research Center, Wuhan University, Wuhan 430071, China yfzhouwhu@163.com 
ZHOU Yun-feng Department of Radio-Chemotherapy, Zhongnan Hospital and Cancer Research Center, Wuhan University, Wuhan 430071, China  
XIE Cong-hua Department of Radio-Chemotherapy, Zhongnan Hospital and Cancer Research Center, Wuhan University, Wuhan 430071, China  
熊杰 Department of Radio-Chemotherapy, Zhongnan Hospital and Cancer Research Center, Wuhan University, Wuhan 430071, China  
鲍洁 Department of Radio-Chemotherapy, Zhongnan Hospital and Cancer Research Center, Wuhan University, Wuhan 430071, China  
周福祥 Department of Radio-Chemotherapy, Zhongnan Hospital and Cancer Research Center, Wuhan University, Wuhan 430071, China  
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Abstract::
      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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