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Korean Journal of Obstetrics & Gynecology 1999;42(5):1108-1116.
Published online January 1, 2001.
Growth Inhibition of Cervical Cancer Cell by Lipofectin with Adeno Associated Viral Plasmid [pAAVCMVp53/lipofectin].
S Y Seo, J Y Lee, W S An, G N Jo, Y J Han, J M Lee, S E Namkoong, H Y Lee, S J Kim, S P Kim, D J Kim, Y S Park, J K Kim, J M Yang, S H Park
1Dept of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea.
2Catholic Research Institute of Medical Science.
3Department of Statistics, College of Medicine, The Catholic University of Korea.
4Department of medical technology, College of Health Science, Yonsei University.
5College of Pharmacy, Seoul National University.
6Department of Biology, Sogang University.
7Genetic Engineering Research Institute, Korea Institute of Science and Technology.
Abstract
PURPOSE
Cervical cancer is strongly associated with human papilloma virus [HPV] and the transforming viral genes E6 and E7 are steadily expressed by the tumor cells. To evaluate the transfection efficiency of pAAVCMVp53 by lipofectin and explore the growth inhibitory potential of an pAAVCMVp53 in various cervical cancer cell lines, we constructed pAAVCMVp53. METHODS: CaSki, SiHa, HeLa, HeLaS3, C33A and HT3 cervical cancer cell lines were used for evaluate the growth inhibitory effect of pAAVCMVp53 transferred by lipofectin. These cells were cultured for 6 days, we divided transfection groups into five groups, control, pAAVCMVp53, lipofectin, pAAVCMVp53/lipofectin and pAAVCMV/lipofectin. Cell proliferation was checked by cell counting after Trypan blues staining, cell viability assay by ELISA method stained by neutral red, MTT assay and [3H] Thymidine incorporation assay. And Western blot was performed for confirming p53 gene expression after 1, 3, 5 days. For the statistical analysis one-way ANOVA F-test, multiple comparison Tukey method was used. RESULTS: The third day after gene transfection showed the highest expression of p53 protein. Transfection efficiency was about 10%~37.5%. pAAVCMVp53 inhibited cell growth with lipofection by cell counting after 6 day culture about 60.8% CaSki and 58.5% in SiHa cells, 35.5% in HeLa and 67.4% in HeLaS3, 53.1% in C33A and 86.9% in HT3 compared to control cells. CONCLUSION: These data suggest that transfection of cervical cancer cells with pAAVCMVp53 with lipofectin is a potential novel approach to the gene therapy of cervical cancer.
Key Words: p53, pAAVCMVp53, cervical cancer, gene therapy


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