Establishment of a Hepatocellular Carcinoma Cell Line Expressing Dual Reporter Genes: Sodium Iodide Symporter (NIS) and Enhanced Green Fluorescence Protein (EGFP)
作者： Byeong-Cheol Ahn , Sang-Woo Lee , Jae-Tae Lee , Jeong-Soo Yoo , Won-Jung Kwak
DOI:
关键词: Fluorescence microscope 、 Cell 、 Transfection 、 Gene 、 Sodium-iodide symporter 、 Molecular biology 、 Reporter gene 、 Biology 、 Green fluorescent protein 、 Genetic enhancement
摘要: Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular studies such as therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), the functionalities were evaluated in vivo by nuclear optical imaging. Materials Methods: A pRetro-PN vector was after separating NIS from pcDNA-NIS. RSV-EGFP-WPRE fragment separated pLNRGW cloned into vector. The final named pRetro-PNRGW. human (HepG2) cells transfected retrovirus containing EGFP (HepG2-NE). Expression confirmed RT-PCR, radioiodine uptake efflux studies. RT-PCR microscope. HepG2 HepG2-NE implanted shoulder hindlimb nude mice, then image, gamma camera image I-124 microPET undertaken. Results: successfully constructed. showed mRNA expression. About 50% fluorescence. iodine NIS-expressed about 9 times higher than control. In study, min. xenograft high signal-to-background fluorescent spots iodine-uptake compared to those xenograft. Conclusion: could be used a potential either therapeutic or gene.
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