| LIAO Zheng-kai,ZHOU Yun-feng,ZHOU Fu-xiang,et al.hTERT promoter mediating gene therapy in laryngeal squamous carcinomas cells in vitro[J].Chinese Journal of Radiological Medicine and Protection,2007,27(2):109-112 |
| hTERT promoter mediating gene therapy in laryngeal squamous carcinomas cells in vitro |
| Received:August 28, 2006 |
| DOI： |
| KeyWords:Gene therapy 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项目(2006ABC009) |
| Author Name | Affiliation | E-mail | | LIAO Zheng-kai | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | | | ZHOU Yun-feng | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | yfzhouwhu@163.com | | ZHOU Fu-xiang | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | | | 骆志国 | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | | | 熊杰 | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | | | 鲍洁 | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | | | 谢丛华 | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | | | 刘诗权 | Department of Radio-Chemotherapy, Cancer Research Center Zhongnan Hospital, Wuhan University, Wuhan 430071, China | |
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| Abstract:: |
| Objective To investigate the relationship among hTERT promoter activity, hTERT mRNA expression, and telomerase activity(TA) in laryngeal squamous carcinomas cell lines, and to evaluate the usefulness of hTERT promoter mediated gene therapy. Methods After plasmids pGL3-hTERTp were transfected, hTERT promoter activity, hTERT mRNA expression and TA were determined by luciferase assay, RT-PCR and TRAP-PCR-ELISA, respectively. Plasmid phTERTp-HRP was constructed and transfected, HRP expression was determined by RT-PCR and competent peroxidase activity was confirmed by enzyme activity assay. The cytotoxicity and radiosensitivity of phTERTp-HRP/IAA were determined by clonogenic assay. Results The relative levels of hTERT promoter activity, hTERT mRNA expression and TA in Hep2R cells were 1.37-fold, 1.43-fold and 1.81-fold compared with Hep2R cells. hTERT promoter activity was closely associated with hTERT mRNA expression and TA levels(P<0.01). In phTERTp-HRP transfected cells, the HRP mRNA expression level and HRP activity in Hep2R cells were 2.1-fold and 1.8-fold compared with Hep2R cells. After IAA incubation, the sensitizer enhancement ratio(SERSF2) was 1.24(Hep2R cells) and 1.20(Hep 2cells) ,the parameter α of with or without IAA incubation were 0.090,0.020(Hep2R)and 0.099,0.042(Hep2). Conclusions hTERT promoter is applicable in mediating gene therapy in different radiosensitive laryngeal squamous carcinomas cells. hTERTp-HRP/IAA gene therapy may be a promising supplementary method for radiotherapy of laryngeal squamous-cell carcinomas. |
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