Nucl Med Mol Imaging.  2007 Jun;41(3):226-233.

Establishment of a Hepatocellular Carcinoma Cell Line Expressing Dual Reporter Genes: Sodium Iodide Symporter (NIS) and Enhanced Green Fluorescence Protein (EGFP)

• Kwak W
• Koo BC
• Kwon MS
• Lee YJ
• Lee HY
• Yoo J
• Kim T
• Chun KS
• Cheon GJ
• Lee SW
• Ahn BC
• Lee J

Abstract

PURPOSE: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging.
MATERIALS AND METHODS
A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken.
RESULTS
The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, T(1/2) of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft.
CONCLUSION
A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.

Keyword

dual reporter gene; HepG2 cell; NIS; EGFP

MeSH Terms

Animals
Carcinoma, Hepatocellular*
Cell Line*
Clone Cells
Fluorescence*
Gamma Cameras
Genes, Reporter*
Genetic Therapy
Hep G2 Cells
Heterografts
Hindlimb
Humans
Iodine
Ion Transport*
Mice
Mice, Nude
Molecular Imaging
Optical Imaging
Retroviridae
RNA, Messenger
Shoulder
Sodium Iodide*
Sodium*
Iodine
RNA, Messenger
Sodium
Sodium Iodide