| Effects of Wild - type p53 Gene Transfection into Human Colon Cancer Cell Line |
| Hyun Ok Kim, Woo Ho Kim, Soo In Bae, He Won Lee, Chong Jai Kim, Sung Youl Hong, Yong Il Kim |
1Department of Pathology, Seoul National University College of Medicine, Seoul, Korea.
2Department of Biotechnology, Sungkyunkwan University, Suwon, Korea.
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| ABSTRACT |
PURPOSE:
In colon cancer, the most frequent genetic alteration is found in p53 tumor suppressor gene residing on the short arm of chromosome 17. In order to investigate the significance of wild-type p53, we transfected wild type p53 into human colon cancer cell lines and analysed tbeir biologic effects.
MATERIALS AND METHODS:
For analysis of p53 status in cell lines, polymerase chain reaction-single stranded confonnation polymorphism (PCR-SSCP), PCR-direct sequencing and Western blot analysis were employed. Transient transfection with liposome-p53 complex was followed by cell biologic assay.
RESULTS:
We found that twelve of fifteen human colon cancer cell lines showed mutation of p53 by PCR-SSCP method. These results almost corresponded to p53 protein accumulations assessed by Westem blot using PAbl801. After transfection with lipafect- AMINE and wild type p53 complex on p53 mutant type cell line (LS1034), viability was reduced to 17.9%, and invasiveness was reduced to 37.3%. Morphologically, wild type p53 transfected cells showed lumen formation and apoptosis after induction of differentiation by Matrigel.
CONCLUSION:
Wild type p53 transfection into p53 mutated colon cancer ceil line resulted in restoration of tumor suppressor effect of p53, and this model would be one of the experimental systems for p53-based gene therapy. |
| Key words:
Colon cancer celi lines;p53 tumor suppressor gene;Liposome;Transfection |
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