Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul, Korea.
Int J Radiat Biol. 2011 Dec;87(12):1182-8. doi: 10.3109/09553002.2011.630440. Epub 2011 Nov 9.
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.
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.
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.
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.
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。
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。
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)。
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。