Journal of Pharmaceutical Investigation

The Korean Society of Pharmaceutical Sciences and Technology (한국약제학회)
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EBV-Based Plasmid Encoding HSV-TK for Cytocidal Gene Therapy

HSV-TK 유전자를 암호화하는 EBV 유래 플라스미드를 이용한 유전자 치료

  • Oh, Sang-Taek (Research Institute of Immunobiology, Catholic Research Institutes of Medical Science, Catholic University) ;
  • Min, Kyoung-Ah (Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University) ;
  • Kim, Chong-Kook (Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University) ;
  • Lee, Suk-Kyeong (Research Institute of Immunobiology, Catholic Research Institutes of Medical Science, Catholic University)
  • 오상택 (가톨릭대학교 의과학연구원 면역생물학연구소) ;
  • 민경아 (서울대학교 약학대학) ;
  • 김종국 (서울대학교 약학대학) ;
  • 이숙경 (가톨릭대학교 의과학연구원 면역생물학연구소)
  • Published : 2003.12.20
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Abstract

Herpes simplex virus (HSV) thymidine kinase (TK) has been widely used for suicidal gene therapy in combination with nucleoside analogs such as ganciclovir (GCV). The use of HSV-TK is limited due to the side effect of GCV at high concentrations. Previous studies showed that stable transfectants of mutant HSV-TK with enhanced affinity to GCV were killed at lower GCV concentrations. In this study, we tested whether mutant HSV-TK can provide enhanced suicidal effect when transiently transfected with Epstein-Barr virus (EBV)-based plasmid. EBV-based plasmid which contains OriP and EBNA-1 sequence is well known for a stable episomal maintenance in human cells. Optimal transfection condition was assessed for SNU-638 gastric cancer cell line using polyetylnimine (PEI). Maximum transfection efficiency was achieved when DNA:PEI was 1:3 (w/v). Cytotoxicities of mutant and wild type HSV-TK were compared before and after partially selecting transfected cells. The cells were sensitive to $100\;{\mu}g/ml$ hygromycin. Following GCV treatment, more cells were killed after hygromycin selection than before selection. The mutant HSV-TK showed enhanced cytotoxicity compared with the wild type HSV-TK. Our results suggest that the EBV-based plasmid encoding mutant HSV-TK may be useful to treat the diseases caused by uncontrolled cell proliferation such as cancer and rheumatoid arthritis.

Keywords

References

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