| ZHAO Bao_feng,TIAN Mei,LEI Hong_wei.Cloning of smac gene and its overexpression effects on radiosensitivity of HeLa cells to γ-rays[J].Chinese Journal of Radiological Medicine and Protection,2006,26(5):456-459 |
| Cloning of smac gene and its overexpression effects on radiosensitivity of HeLa cells to γ-rays |
| Received:February 01, 2006 |
| DOI： |
| KeyWords:Smac gene Radiosensitivity γ-ray HeLa cells |
| FundProject:中国博士后基金资助项目(2005038364);兰州重离子加速器国家实验室基金资助 |
| Author Name | Affiliation | E-mail | | ZHAO Bao_feng | National Institute for Radiological Protection, China CDC, Beijing 100088, China | suxu@nirp.cn | | TIAN Mei | National Institute for Radiological Protection, China CDC, Beijing 100088, China | | | LEI Hong_wei | National Institute for Radiological Protection, China CDC, Beijing 100088, China | | | 苏旭 | National Institute for Radiological Protection, China CDC, Beijing 100088, China | |
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| Abstract:: |
| Objective To clone smac gene and construct eukaryocytic expression vector pcDNA3.1/smac. The smac gene was transfected into HeLa cells to explore the effects of over-expression of extrinsic smac gene on radiosensitivity to γ-rays of HeLa cells. Methods The full-length smac gene was amplified from total RNA of HeLa cells by RTPCR. The RTPCR product was ligated with the vector pcDNA3.1 and sequenced. The correct pcDNA3.1/smac was transfected into HeLa cells. The expression of smac gene was tested by RTPCR and Western blot. The cellular growth inhibition rates were evaluated by MTT 48 hours after irradiation with different doses of γ-rays. Results Recombinant eukaryocytic expression vector pcDNA3.1/smac was successfully constructed. RTPCR and Western blot results indicated that the expression of smac gene of HeLa/smac cells was significantly enhanced compared with the expression of smac gene of HeLa/pcDNA3.1 and HeLa cells. 48 hours after different doses of γ-ray irradiation was significantly higher in pcDNA3.1/smac transfected HeLa/smac cells than those of non-transfected HeLa cells or pcDNA3.1 transfected HeLa/pcDNA3.1 cells, inhabitation rates were 38.85%,17.64%and 20.32%,respectively. Conclusions smac gene was successfully cloned. Extrinsic smac gene over-expression could significantly enhance radiosensitivity to γ-ray of HeLa cells,which would herald a new approach to improve radiosensitivity of cervical cancer. |
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