Growth Inhibitory Effect of Human Cervical Cancer Cells with the Direct Transfer of Liposome Complexed Recombinant pRcCMVp53

Kim, J C; Seo, S Y; Shin, B Y; An, W S; Han, Y J; Lee, J M; Namkoong, S E; Lee, H Y; Kim, S J; Kim, S P; Kim, D J; Park, Y S; Kim, J K; Yang, J M; Park, S H

Abstract

The Catholic University of Korea,1 Catholic Research Institutes of Medical Science,2 Department of Biostatistics, The Catholic University of Korea,3 Dept of medical technology, College of Health Science, Yonsei University,4 College of Pharmacy, Seoul National University,5 Dept of Biology, Sogang University,6 Genetic Engineering Research Institute, Korea Institute of Science and Technology7 Purpose: We investigated cell growth inhibitory effect of the wild type p53 gene into the cervical cancer cells via the recombinant p53 plasmid, pRcCMVp53 with lipofectin. METHOD: Inhibition of the growth of cervical cells as determined by a cell count assay. The cells were inoculated at density of 104 cells/well in each 12 well plate, 24 hrs before infection. At each indicated point, cells in three wells on each well plate were trypsinized and counted. The mean cell counts of triplicated well after transfection at day 1~6 was checked. Inhibition of the growth of cervical cells were checked by ELISA assay, MTT assay. The cells inoculated at densities of 5x103/well in each 96 plate 24 hrs before infection. At each indicated point cells of three wells were transfected for 6 days, harvested and counted by liquid scintillation counter the mean cpm per triplicate wells were plated for ELISA and MTT assay. One-way ANOVA F-test and multiple comparison Tukey method was used for statistical analysis. RESULT: Inhibition of the growth of cervical cells in cell count showed CaSki[89%], SiHa[59.2%], HeLa[86%], HeLaS3[78%], C33A[91%] and HT3[74%]. ELISA assay showed CaSki[70%], SiHa[53%], HeLa[73%], HeLaS3[50%], C33A[67%] and HT3[73%]. MTT assay showed HeLa[33%], HeLaS3[28%], SiHa[75%], CaSki[38%], C33A[33%] and HT3[53%].
CONCLUSION
These results indicate that transfection of cervical cancer cells with the wild type p53 gene via pRcCMVp53 with lipofectin is a potential novel approach to the gene therapy of cervical cancer.