단순 헤르페스 제 1형 티미딘 키나제 유전자 이입 간암세포주에서 방사표지 IVDU와 IVFRU의 섭취 평가 (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 |
이태섭1,4, 최태현1,4, 안순혁1, 우광선1, 정위섭1, 권희충3, 오옥두4, 최창운1,2, 임상무1,2., |
Tae Sup Lee,1,4 M.S., Tae Hyun Choi1,4 M.S., Soon Hyuk Ahn1, M.S., Kwang Sun Woo1,B.A., Wee Sup Jeong1, B.A., Hee Chung Kwon3, Ph.D., Ok Doo Awh4 Ph.D, Chang WoonChoi1,2 M.D., Sang Moo Lim1,2 M.D. |
Affiliation |
원자력의학원 싸이클로트론응용연구실1, 핵의학과2, 분자종양학연구실3, 연세대학교 보건과학대학 임상병리학과4. Laboratory of Cyclotron Application1, Department of Nuclear Medicine2, Laboratory of Molecular Oncology3,Korea Institute of Radiological and Medical Sciences(KIRAMS), Seoul, Korea; Department of Medical Laboratory Science, College of Health Science, Yonse |
Abstract |
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. |
Keyword |
Radiolabeled IVDU and IVFRU, Herpes simplex virus type 1 thymidine kinase(HSV1-tk), Geneimaging, Gene therapy |
Full text Article |
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