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使用人甲状腺钠碘转运体和单体红色荧光蛋白融合报告基因的伽马相机和光学成像在小鼠模型中的应用。

Gamma camera and optical imaging with a fusion reporter gene using human sodium/iodide symporter and monomeric red fluorescent protein in mouse model.

机构信息

Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul, Korea.

出版信息

Int J Radiat Biol. 2011 Dec;87(12):1182-8. doi: 10.3109/09553002.2011.630440. Epub 2011 Nov 9.

Abstract

PURPOSE

Multimodality imaging contributes to the activation of translational research by compensating for its weak points. Herein, we developed a noninvasive dual-reporter gene system for nuclear and optical imaging.

MATERIALS AND METHODS

We constructed a fusion reporter vector concurrently expressing the human sodium/iodide symporter (hNIS) and monomeric red fluorescent protein (mCherry), and evaluated the function of this fusion reporter system under in vitro and in vivo conditions.

RESULTS

The expression of hNIS/mCherry fusion gene was confirmed in transfected cells using reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. As the numbers of cells increased, the fluorescence and 125I uptake increased in the hNIS/mCherry-transfected cells, and a high correlation between fluorescence intensity and radioactivity was noted. The fluorescence intensities and radioactivity signals were also well-correlated in HT-29-hNIS/mCherry tumors (R2=0.9304) in in vivo fluorescence and gamma camera imaging.

CONCLUSIONS

The dual-reporter imaging method using hNIS and mCherry genes reflected tumor extent as well as viable cell numbers, and correlated well with one another. This suggests that the hNIS/mCherry dual-reporter system can be a useful tool for multi-modal imaging.

摘要

目的

多模态成像通过弥补其弱点为转化研究提供了助力。在此,我们开发了一种用于核和光学成像的非侵入性双报告基因系统。

材料与方法

我们构建了一个融合报告载体,同时表达人钠/碘转运体(hNIS)和单体红色荧光蛋白(mCherry),并在体外和体内条件下评估了该融合报告系统的功能。

结果

通过逆转录聚合酶链反应(RT-PCR)和 Western blot 确认了转染细胞中 hNIS/mCherry 融合基因的表达。随着细胞数量的增加,hNIS/mCherry 转染细胞中的荧光和 125I 摄取增加,并且荧光强度与放射性之间存在高度相关性。在体内荧光和伽马相机成像中,HT-29-hNIS/mCherry 肿瘤中的荧光强度和放射性信号也具有很好的相关性(R2=0.9304)。

结论

使用 hNIS 和 mCherry 基因的双报告基因成像方法反映了肿瘤范围和存活细胞数量,并彼此高度相关。这表明 hNIS/mCherry 双报告系统可以成为多模态成像的有用工具。

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