| Li Wei,Tan Jian*,Wang Peng,Li Ning,Li Chengxia.Conditionally replicative adenovirus under the control of glial fibrillary acidic protein and human telomerase reverse transcriptase dual-promoters direct sodium iodide symporter expression for malignant glioma radioiodine therapy[J].Chinese Journal of Radiological Medicine and Protection,2014,34(1):3-7 |
| Conditionally replicative adenovirus under the control of glial fibrillary acidic protein and human telomerase reverse transcriptase dual-promoters direct sodium iodide symporter expression for malignant glioma radioiodine therapy |
| Received:May 14, 2013 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.01.002 |
| KeyWords:Conditionally replicative adenovirus hNIS gene hTERT promoter GFAP promoter Glioma |
| FundProject:国家自然科学基金（81301244，81171372）；天津市应用基础及前沿技术研究计划（12JCZDJC26000） |
| Author Name | Affiliation | E-mail | | Li Wei | Department of Nuclear Medicine, Tianjin Medical University Hospital, Tianjin 300052, China | | | Tan Jian* | Department of Nuclear Medicine, Tianjin Medical University Hospital, Tianjin 300052, China | tanpost@163.com | | Wang Peng | Department of Nuclear Medicine, Tianjin Medical University Hospital, Tianjin 300052, China | | | Li Ning | Department of Nuclear Medicine, Tianjin Medical University Hospital, Tianjin 300052, China | | | Li Chengxia | Department of Nuclear Medicine, Tianjin Medical University Hospital, Tianjin 300052, China | |
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| Abstract:: |
| Objective To explore the possibility of using 131I as a targeted therapy method for malignant glioma by infecting U87 and U251 cells with conditionally replicative adenovirus Ad-Tp-E1a-Gp-NIS. Methods Human telomerase reverse transcriptase (hTERT) promoter and glial fibrillary acidic protein (GFAP) promoter were cloned and their transcriptional activities were detected by luciferase assay. The conditionally replicative adenovirus Ad-Tp-E1a-Gp-NIS was constructed, purified, and transfected into U87 and U251 glioma cells. For these transfected cells, the selective replication ability was evaluated by plaque forming assay, and protein expression was detected by Western blot assay. 125I-iodide uptake and exflux, the clonoy formation of 131I-iodide treated cells were also measured. Results Transcriptions activity of the GFAP and hTERT promoters was 59.75%-62.10% (F=11.89, P<0.01) in U87 cells and 37.31%-49.00% (F=5.87, P<0.05) in U251 cells. The Ad-Tp-E1a-Gp-NIS could be selectively replicated and the hNIS gene was successfully expressed in the hTERT-positive and GFAP-positive glioma cells which showed two protein bands with relative molecular mass of 120×103 and 49×103 in Western blot assay. After infection with Ad-Tp-E1a-Gp-NIS, the cell ability of 125I uptake was increased by 78.80 (F=2 914.58, P<0.01) and 92.48 (F=2 275.91, P<0.01) times in U87 and U251 cells, respectively. The GFAP-negative MRC-5 cells could not take in 125I. The in vitro clonogenic assay indicated that, after 131I treatment, more than 90% of the transfected cells were killed, while only about 65% (t=11.73-78.33, P<0.01) of control cells were killed. Conclusions The Ad-Tp-E1a-Gp-NIS has a good ability in selective replication and the enhancement of antitumor therapy effect by increasing tumor-specific iodide uptake in malignant glioma cells. |
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