125I uptake in U251 glioma cell co-transfected with the human sodium/iodide symporter and the human thyroperoxidase
10.3760/cma.j.issn.0253-9780.2010.06.011
- VernacularTitle:hTPO和hNIS共转染胶质瘤细胞U251介导放射性碘摄取的研究
- Author:
Bei, WU
;
Jian, TAN
;
Lei, LONG
;
Wei, LI
- Publication Type:Journal Article
- Keywords:
Glioma;
Tumor cells,cultured;
Transfection;
Iodide peroxidase;
Sodium/iodide symporter;
Iodine radioisotopes
- From:Chinese Journal of Nuclear Medicine
2010;30(6):395-399
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the iodide uptake by U251 glioma cell lines which were transfered with both human sodium/iodide symporter (hNIS) and human thyroperoxidase (hTPO) genes. Methods Recombinant adenosine virus AdTPO was constructed through cloning, recombination, packaging and amplifying. The viral titers were calculated after purification. The protein expression of AdTPO was tested by Western-Blotting and the recombinant plasmids PcDNA3. 1/hNIS were constructed. After hNIS gene was transfected into human glioma cell lines U251 through liposome, the cell lines with stable hNIS expression (hNIS-U251) selected by G418 antibiotics were defined as hNIS-U251 group. Then, hTPO was transducted into hNIS-U251 with adenosine virus (AdTPO-hNIS-U251 group). U251 cells with no plasmid were used as the control group (U251). Cultured cells from each group were studied for 125I uptake as well as 125I efflux rate. Student-Newman-Keuls in multiple range test was used. Results AdTPO-hNIS-U2.51 with stable expression was successfully established by transfecting hNIS and hTPO genes into human glioma cell lines. The 125I uptake by AdTPO-hNIS-U251, hNIS-U251 and U251 cell lines was (74 647.53 ±3605.88), (55 769.96 ±4353.26) and ( 507.67 ± 57.69 ) counts/min, respectively ( F = 836. 17, P ＜ 0.05 ). The uptake compacity by AdTPO-hNIS-U251 was 147 fold higher than that by U251 (q =55.64, P＜0.01 ) and 1.3 fold higher that by hNIS-U251 (q = 14. 17, P ＜0.01 ). 125I efflux rate was prolonged in AdTPO-hNIS-U251 group and its effective half time was 13 min. Conclusion Enhanced 125I uptake by the human glioma cell lines can be achieved with combined transfection of hNIS and hTPO genes.
