Purpose: We investigated the time-course expression patterns of p53 and E6 on cervical cancer cells to obtain a molecular level understanding of cell-dependent tumor growth suppression effects of recombinant adenovirus expressing p53 in vitro and in vivo.
Methods: Four human papillomavirus (HPV)-infected human cervical cancer cell lines (HPV 16-positive cells, CaSki and SiHa cells; and HPV 18-positive cells, HeLa and HeLaS3 cells) were used. Also, HPV negative C33A and HT3 cell line that has a mutation on p53 gene were used. After infection with AdCMVp53, the cell growth inhibition was studied via cell count assay, MTT assay, and Neutral red assay. After transfecting AdCMVp53 and AdCMVLacZ into the cancer cells-xenografted nude mice, antitumor effects were investigated for 1 month, respectively.
Results: For each cervical cancer cell, IC50 was as follows; CaSki (68.5 multiplicity of infection, or MOI), SiHa (43.5 MOI), HeLa (31 MOI), HeLaS3 (42 MOI), C33A (21 MOI), and HT3 (62 MOI). In particular, complete inhibition of cell growth was observed at 125 MOI in both CaSki and SiHa cells. However, the complete inhibition was detected at 62.5 MOI in HeLa and HeLaS3. In contrast, at these MOI, no suppression of cell growth was observed when cells were infected with recombinant adenovirus expressing beta-gal as a negative control. The levels of p53 protein were notably expressed in CaSki and HeLa more than in SiHa and HeLaS3 on days 2 and 4. However, the p53 was only detected in HeLaS3 on day 6. In contrast, p53 expression was continually maintained in C33A and HT3 during the same periods. After transfection AdCMVp53 into CaSki- and SiHa-xenografted nude mice, the size of tumor was remarkably decreased in SiHa cells as compared to AdCMVLacZ transfection.
Conclusion: The adenovirus-mediated p53 gene transfection was done effectively in vitro and in vivo. Also, the antitumor effects were accomplished via differential role of p53-specific apoptotic cell death, which is dependent upon the cervical cancer cell line.