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
10.3760/cma.j.issn.0254-5098.2014.01.002
- VernacularTitle:hTERT和GFAP双启动子条件复制腺病毒介导放射性碘治疗脑胶质瘤的实验研究
- Author:
Wei LI
;
Jian TAN
;
Peng WANG
;
Ning LI
;
Chengxia LI
- Publication Type:Journal Article
- Keywords:
Conditionally replicative adenovirus;
hNIS gene;
hTERT promoter;
GFAP promoter;
Glioma
- From:
Chinese Journal of Radiological Medicine and Protection
2014;34(1):3-7
- CountryChina
- Language:Chinese
-
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-E1 a-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.
