| CHEN Zhi-guo,YANG Ye-peng,ZHOU Mei.Cytocidal efficacy of γ-rays combined with HSV-tk/GCV gene therapy on Lewis lung carcinoma cells in vitro[J].Chinese Journal of Radiological Medicine and Protection,2005,25(6):509-513 |
| Cytocidal efficacy of γ-rays combined with HSV-tk/GCV gene therapy on Lewis lung carcinoma cells in vitro |
| Received:December 29, 2004 |
| DOI： |
| KeyWords:γ-rays HSV-tk Gene therapy Lewis lung carcinoma cells Bystander effect |
| FundProject:国家自然科学基金资助项目(30470523) |
| Author Name | Affiliation | E-mail | | CHEN Zhi-guo | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | yangyepeng@bjmu.edu.cn | | YANG Ye-peng | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | | | ZHOU Mei | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | | | 占晓海 | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | | | 沈嵚剑 | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | | | 沈磊 | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | | | 李五岭 | Department of Radiation Medicine, School of Basic Medical Sciences, Peking University, Beijing 100083, China | | | 王新娟 | 100083, 北京大学生物化学与分子生物学系 | |
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| Abstract:: |
| Objective To observe cytocidal efficacy of γ-rays combined with HSV-tk/GCV gene therapy on murine Lewis lung carcinoma cells in vitro and to explore the mechanism of the combined effects. Methods HSV-tk gene packaged with Lipofectin was stably transfected into Lewis lung carcinoma cells. HSV-tk mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR). Hoechst/PI vital staining and flow cytometry were used to measure apoptosis induced by the individual and the combined treatment of γ-rays and HSV-tk/GCV system. Cell survival curves were determined by colony formation method to evaluate changes of radiosensitivity of the cells. Moreover, bystander effect of the HSV-tk/GCV system and influence of γ irradiation on the bystander effect were detected. Results The HSV-tk/GCV system showed a selective cytocidal effect on the Lewis cells transfected with HSV-tk gene (Lewis-Hytk cells). The apoptosis rate and necrosis rate of the cells treated by combined modality was significantly higher than the sum of those of the cells treated separately by these two individual modalities. A dose of 0.1 μg/ml GCV could obviously raise the radiosensitivity of Lewis-Hytk cells (SER=1.47). It was also noted that γ-rays could enhance the bystander effect of HSV-tk/GCV system and the medium from cultured Lewis-Hytk cells treated by combined modality showed a clear cell-killing effect on untreated Lewis cells. Conclusions γ-rays combined with HSV-tk/GCV system can produce synergic cell-killing effects on Lewis lung carcinoma cells. γ irradiation can enhance the bystander effect of HSV-tk/GCV system, which may be one of the mechanisms that results in the synergism. |
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