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Korean Journal of Obstetrics & Gynecology 2004;47(5):870-879.
Published online May 1, 2004.
Changes of Transfection Efficiencies of Plasmid LacZ Genes in Human Gynecologic Cancer Cell Lines.
Byung Hoon Kim, Su Mi Bae, You Jin Han, Hyeon Seung Lee, Yoon Sung Jo, Keun Ho Lee, Soo Young Hur, Jae Hoon Kim, Young Wook Kim, Duck Yong Ro, Joon Mo Lee, Sung Eun Namkoong, Dong Jae Kim, Chong Kook Kim, Woong Shick Ahn
1Catholic Research Institutes of Medical Science, The Catholic University of Korea, Korea.
2Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea, Korea.
3Department of Statistics, College of Medicine, The Catholic University of Korea, Korea.
4College of Pharmacy, Seoul National University, Korea.
Abstract
OBJECTIVE
The transfection efficiencies of gynecologic cancer cell lines were investigated by different mediated transfection methods using recombinant LacZ plasmid (pRcCMVLacZ and pAAVCMVLacZ). METHODS: In this study, the gynecologic cancer cell lines were used CaSki, SiHa (cervical, HPV16+, wild type p53 gene), HeLa, HeLa S3 (cervical, HPV18+, wild type p53 gene), C33A, HT3 (cervical, HPV-, p53 mutant), HckE6/E7 (cervical, HPV16 immortalized keratocyte), PA-1 (ovary, wild type p53), SKOV-3, A2774 (ovary, p53del) and OVCAR-3 (ovary, p53 mutant). The pRcCMVLacZ and pAAVCMVLacZ plasmid transfection were performed by using liposome system such as Ca2+-phosphate, Fugen6(TM), Lipofection(TM), Lipogen(TM) and N-stearyl lactobionamide (N-SLBA) with X-gal staining. The LacZ gene was used the reporter gene for the transfection efficiencies evaluation. RESULTS: Each of cell lines were showed different transfection efficiencies by Ca2+-phosphate, Fugen6(TM), Lipofectin(TM), Lipogen(TM) and N-SLBA. Each of cell were revealed that HeLa S3, HT3 and A2774 were high transfection efficiency using the pRcCMVLacZ by the Lipogen(TM), SiHa, HeLa, QGU, OVCAR-3 and PA-1 were high efficiency using the pAAVCMVLacZ by Lipofectin(TM), CaSki was high efficiency using the pRcCMVLacZ by the Lipogen(TM), A2774 and Cx16.2 were high efficiency using the pRcCMVLacZ by the Lipofectin(TM), SKOV-3 and HkcE6/E7 were high efficiency using pAAVCMVLacZ by the Lipogen(TM). CONCLUSION: As a result, We proved that each of cell lines differed trasnfection efficiencies according to mediated transfection and recombinant LacZ plasmid style. Above all, Lipofectin(TM) mediated transfection was showed high efficiency at the most of cell lines.
Key Words: pRcCMALacZ, pAAVCMVLacZ, Gynecology cancer, Gene transfection, X-gal staining


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