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Korean Journal of Obstetrics & Gynecology 2004;47(2):258-263.
Published online February 1, 2004.
Development of Trans Attack Anticancer Gene Vaccine Interact in HPV Oncoprotein Expression Inhibition.
Hee Joong Lee, Sung Ha Lee, Hyun Jung Kim, Jae Hoon Kim, Young Ok Lew, Jin Ok Lee, Young Wan Kim, Woong Shick Ahn, Chong Kook Kim, Jun Mo Lee, Dae Hoon Kim, Sung Eun Namkoong, Kim Dou Kang
1Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea.
2Catholic Research Institutes of Medical Science, College of Medicine, The Catholic University of Korea.
3College of Pharmacy, Seoul National University, Korea.
Abstract
OBJECTIVE
Adeno-associated virus Rep 78 protein is known to inhibit the promoter site of several onco-genes and viral gene, including the human papillomavirus type 16 E6 transforming genes. In this study, we investigated AAV Rep 78 mediated inhibition of HPV 16 E6 promotor activity. METHODS: pcDNA3.1/V5/His-Topo vector, cloned by AAV mediated Rep 78, is transfected into cervical cancer cell line (Caski). After that, we confirmed HPV16 derived E6 expression and cell growth inhibition. RESULTS: Transfection rate of Rep78 GFP-vector, approximately from 30 to 60 per-cent, is highly expressed at first day. But E6 expression is lower at this day. The growth of CaSki and HeLa cervical cancer cell lines was inhibited by Rep78 (p<0.05). But, the other cervical cancer cells were unaffected by Rep78 transfection. CONCLUSION: In spite of the high Rep78 transfection efficiency and expression rate, we could not show the cervical cancer cell growth inhibition. In our data, long term expression of Rep78 strategy is needed for cervical carcinoma gene therapy using adeno-associated virus vector.
Key Words: Rep78, AAV (adeno-associated virus), E6 oncoprotein, HPV
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