In Vitro Uptakes of Radiolabeled IVDU and IVFRU in Herpes Simplex Virus Type-1 Thymidine Kinase (HSV1-tk) Gene Transduced Morris Hepatoma Cell Line.
- Author:
Tae Sup LEE
1
;
Tae Hyun CHOI
;
Soon Hyuk AHN
;
Kwang Sun WOO
;
Wee Sup JEONG
;
Hee Chung KWON
;
Ok Doo AWH
;
Chang Woon CHOI
;
Sang Moo LIM
Author Information
1. Laboratory of Cyclotron Application, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul, Korea. smlim328@kcch.re.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Radiolabeled IVDU and IVFRU;
Herpes simplex virus type;
thymidine kinase (HSV1-tk);
Gene imaging;
Gene therapy
- MeSH:
Animals;
Blotting, Northern;
Blotting, Western;
Cell Line*;
Gene Expression;
Genes, Reporter;
Genetic Therapy;
Herpes Simplex*;
Herpesvirus 1, Human;
Liver Neoplasms, Experimental*;
RNA, Messenger;
Simplexvirus*;
Thymidine Kinase*;
Thymidine*;
Zidovudine
- From:Korean Journal of Nuclear Medicine
2004;38(1):62-73
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: The herpes simplex virus type 1 thymidine kinase gene (HSV1-tk) is an attractive candidate as a reporter gene in noninvasive reporter gene monitoring system. The HSV1-tk gene was chosen as a reporter gene, because it has been extensively studied, and there are appropriate reporter probes, substrates of HSV1-tk gene product, to apply for HSV1-tk gene imaging. We used radiolabeled 5-iodovinyl-2'-deoxyuridine (IVDU) and 5-Iodovinyl-2'-fluoro-2'-deoxyuridine (IVFRU) as reporter probes for HSV1-tk gene monitoring system. MATERIALS AND METHODS: We prepared HSV1-tk gene transduced Morris hepatoma cell line using retroviral vector, MOLTEN containing HSV1-tk gene. And we confirmed the HSV1-tk gene expression by Northern blotting and Western blotting. We compared in vitro uptakes of radioiodinated IVDU and IVFRU to monitor HSV1-tk gene expression in Morris hepatoma cell line (MCA) and HSV1-tk gene tranduced MCA (MCA-tk) cells until 480 minutes. We also performed correlation analysis between percentage of HSV1-tk gene tranduced MCA cell % (MCA-tk%) and uptakes of radiolabeled IVDU or IVFRU. RESULTS: MCA-tk cell expressed HSV1-tk mRNA and HSV1-TK protein. Two compounds showed minimal uptake in MCA, but increased uptake was observed in MCA-tk. IVDU showed 4-fold higher accumulation than IVFRU at 480 min in MCA-tk (p< 0.01). Both IVDU and IVFRU uptake were linearly correlated (R2> 0.96) with increasing MCA-tk%. CONCLUSION: The radiolabeld IVDU and IVFRU showed higher specific accumulation in retrovirally HSV1-tk gene transfected Morris hepatoma cell line. Both IVDU and IVFRU could be used as good substrates for evaluation of HSV1-tk gene expression.
