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Effect of GCV on Neuroblastoma Cell Line Expressed by HSV-TK Gene with Retroviral Vector

Journal of the Korean Pediatric Society 1997;40(12):1719-1724.
Published online December 15, 1997.
Effect of GCV on Neuroblastoma Cell Line Expressed by HSV-TK Gene with Retroviral Vector
Hyun Sang Cho1, Chuhl Joo Lyu1, Yeun Soo Kim2, Tae Soo Kim2, Byung Soo Kim1
1Department of Pediatrics, College of Medicine, Yonsei University, Seoul, Korea
2Yonsei Cancer Center, Cancer Institute, Seoul, Korea
Retroviral Vector에 의해 HSV-TK 유전자가 발현된 신경아세포주의 성장에 Ganciclovir가 미치는 영향
조현상1, 유철주1, 김연수2, 김태수2, 김병수1
1연세대학교 의과대학 소아과학교실
2연세대학교 의과대학 연세 암센터, 암연구소
Abstract
Background
: Gene transfer with vectors derived from murine retroviruses is restricted to cells which are proliferating and synthesizing DNA at the time of infection. Accordingly, selective introduction of genes encoding for susceptibility to otherwise nontoxic drugs(suicide genes) into proliferating tumor may be used to treat cancer. We investigated the efficacy of in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase(HSV-tk) gene followed by administration of the antiviral drug ganciclovir.
Methods
: The LNC/tK vector was transfered in vitro into mouse Neuro 2a cell lines(ATCC) and the transduced cell lines were selected in G-418, 500μg/ml, for 14 days. One×104 cells were cultured in 96 well culture plates in increasing concentrations of ganciclovir for 72 hours. The sesitivity to ganciclivir of these HSV-tk transduced, G-418 selected cells was measured with MTT assay
Results
: The survival of HSV-tk transduced 1×104 neuro 2a cell lines is 103±3.5%, 68±4.2%, 54±3.8%, 17±2.6%, 13±3.1% at the concentration of 0, 0.1, 1.0, 10, 20μg/ml ganciclovir, respectively. And the survival of HSV-tk not transduced 1×104 neuro 2a cell lines is 100±4.5%, 97±5.6%, 104±3.5%, 106±3.8%, 101±4.2%.
Conclusion
: We concluded that in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase gene followed by administration of the antiviral drug ganciclovir is very effective.
Key Words: Retrovirus, HSV-tk, Neuroblastoma


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