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. 2010 Jun;13(6):575-9.
doi: 10.3779/j.issn.1009-3419.2010.06.002.

[Study on the iodine 125 uptake of H460 lung cancer cell line by co-transfection with the human sodium/iodide symporter and the human thyroperoxidase]

[Article in Chinese]
Wei Li  1 , Jian Tan, Lei Long
Affiliations

[Study on the iodine 125 uptake of H460 lung cancer cell line by co-transfection with the human sodium/iodide symporter and the human thyroperoxidase]

[Article in Chinese]
Wei Li et al. Zhongguo Fei Ai Za Zhi. 2010 Jun.

Abstract
in English, Chinese

Background and objective: Lung cancer harms people's health or even lives severely. Especially, the therapy of non-small cell lung cancer (NSCLC) has not been obviously improved for many years. The aim of this study is to transfer the human sodium/iodide symporter (hNIS) and the human thyroperoxidase (hTPO) genes into H460 lung cancer cell line, and to study the uptake ability of iodide after co-transfected hTPO and hNIS gene in cell lines.

Methods: Through cloning, recombination, packaging and amplifying, the recombinant adenosine virus (AdTPO) was constructed. Then the protein expression of AdTPO was tested by Western blot. After transfected hNIS gene into human lung cancer cell line H460 through liposome, stably expressing hNIS gene cell lines (hNIS-H460) selected by G418 antibiotics was determined as hNIS-H460 group. Using AdTPO, hTPO gene was transducted into hNIS-H460, as AdTPO-hNIS-H460 group. H460 cell without hNIS gene was applied as control group (H460). Then, we investigated the 125I uptake assay of the above cells.

Results: We were successful in co-transfecting hNIS and hTPO gene into human lung cell lines H460, and were obtained hNIS and hTPO gene lung cancer cell lines (hNIS-H460 and AdTPO-hNIS-H460). In AdTPO-hNIS-H460, hNIS-H460 and H460, the uptake ability of 125I was (59 637.67 +/- 1 281.13), (48 622.17 +/- 2 242.28) and (1 440.17 +/- 372.86) counts x min(-1). The uptake ability of 125I was 41 fold higher in AdTPO-hNIS-H460 than in blank control H460 (P < 0.01), and 34 fold higher in hNIS-460 than in blank control H460 (P < 0.01), and 1.2 fold higher in AdTPO-hNIS-H460 than in hNIS-H460 (P < 0.01).

Conclusion: The uptake ability of 125I could increase by co-transfected hNIS and hTPO genes into human lung cancer cell lines H460.

背景与目的: 肺癌严重危害人类生命和健康，目前国内外肺癌的发病率和死亡率仍在不断上升。尤其是非小细胞肺癌（non-small cell lung cacer, NSCLC）治疗效果多年来一直没有显著提高。本研究旨在将人甲状腺过氧化物酶（hTPO）基因及人钠碘转运体（hNIS）基因共转入肺癌细胞系后研究其摄碘能力的变化。

方法: 克隆，重组，包装并扩增纯化得到重组腺病毒（AdTPO），测定病毒滴度，Western blot检测重组腺病毒的表达。使用脂质体转染法将hNIS基因转染入人肺癌细胞系H460中，经G418抗生素筛选获得稳定表达hNIS的细胞系hNIS-H460，为hNIS-H460组；使用重组腺病毒将hTPO基因转导入hNIS-H460中，使肺癌细胞系获得hTPO基因，为AdTPO-hNIS-H460组；未转入hTPO和hNIS的细胞为对照组（H460）。然后进行三组稳定表达细胞系的体外摄125Ⅰ实验。三组间两两比较用q检验（Newman-Keuls法）。

结果: AdTPO-hNIS-H460细胞、hNIS-H460细胞和H460细胞所摄取125Ⅰ分别为（59 637.67±1 281.13）、（48 622.17±2 242.28.）和（1 440.17±372.86）计数·min-1。三组间总体具有统计学差异（P < 0.01）。AdTPO-hNIS-H460组较对照组（H460）摄125Ⅰ能力增高约41倍（P < 0.01），hNIS-H460组较对照组（H460）摄碘能力增高约34倍（P < 0.01）。AdTPO-hNIS-H460组较hNIS-H460组高约1.2倍（P < 0.01）。

结论: 将hTPO和hNIS基因共转染至肺癌细胞系H460后，可有效提高H460的摄碘能力。

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Figures

1
1
PacI酶切AdTPO载体 Ad-TPO vector digested with PacI
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2
Western blot结果。M：Marker条带；1：pAdTrack-CMV-TPO转染后细胞系；2：对照组细胞系。 Result of Western blot. M: Marker; 1: lysate of cells transfected with pAdTrack-CMV-TPO; 2: lysate of control cells.
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G418筛选后各细胞系的摄125Ⅰ能力 The 125Ⅰ uptake ability of cell lines after selected by G418 antibiotics
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各组细胞125Ⅰ外流图 125Ⅰ efflux experiments of cell lines
See this image and copyright information in PMC

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