TIAN Mei,ZHAO Bao-feng,RUAN Jian-lei,et al.Study on enhanced expression and anti-tumor effect of soluble TRAIL induced by 12 C6+ heavy ion irradiation[J].Chinese Journal of Radiological Medicine and Protection,2007,27(3):238-240 |
Study on enhanced expression and anti-tumor effect of soluble TRAIL induced by 12 C6+ heavy ion irradiation |
Received:July 31, 2006 |
DOI: |
KeyWords:pEgr-sTRAIL 12C6+ heavy ion Apoptosis Colony formation HeLa |
FundProject:兰州重离子加速器国家实验室资助项目;中国博士后基金资助项目(2005038364) |
Author Name | Affiliation | E-mail | TIAN Mei | National Institute for Radiological Protection, China CDC, Beijing 100088, China | | ZHAO Bao-feng | National Institute for Radiological Protection, China CDC, Beijing 100088, China | | RUAN Jian-lei | National Institute for Radiological Protection, China CDC, Beijing 100088, China | | 李文建 | 中国科学院兰州近代物理研究所 | | 苏旭 | National Institute for Radiological Protection, China CDC, Beijing 100088, China | suxu@nirp.cn |
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Abstract:: |
Objective To construct pEgr-sTRAIL expression vector and investigate the apoptotic effect and survival level of its transient transfer on cervical cancer cell line HeLa followed by different doses of 12C6+ heavy ion irradiation. Methods Human soluble TRAIL was cloned by SOEing method and then was linked with T-vector for sequencing. The correct fragment was inserted into pcDNA-Egr to construct pEgr-sTRAIL recombinant vector which can be activated and induced by ionizing irradiation. Then pEgr-sTRAIL recombinant vector was transferred into HeLa cells under mediation of GeneCompanionTM in vitro. After different doses of 12C6+ heavy ion irradiation, the expression of TRAIL was detected by RT-PCR, the early apoptosis of transfected cells was measured by FACScan and the cell survival level was determined by colony formation assay. Results The expression vector pEgr-sTRAIL was successfully constructed. 12C6+ heavy ion irradiation could enhance the expression of pEgr - sTRAIL at mRNA level. After irradiated by 12C6+ heavy ion, the percentage of apoptotic cells in pEgr-sTRAIL transfected cells was significantly higher than that of non-transfected cells. The rate of colony formation of pEgr-sTRAIL transfected cells was significantly lower. Conclusion 12C6+ heavy ion irradiation combined with pEgr-sTRAIL gene transfer can significantly induce the apoptosis of tumor cells and have strong anti-tumor effect in vitro. |
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