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When comparing SV40 T antigen-transformed human fibroblasts of a younger generation (24 population doubling) and aging stage (58 population doubling), we found that detachment of cells from the culture surface occurred more frequently in aging cells. DNA fragmentation and chromatin condensation which are typical findings of apoptosis occurred more frequently in aging cells as compared to cells of a younger generation. There is no increase in the p53 level or decrease in the SV40 T antigen level in aging cells as compared to cells of a younger generation. Retinoic acid treatment which can effectively suppress p21 gene expression did not prevent apoptosis. These findings indicate that apoptosis that occurs due to aging-transformed human fibroblasts is mediated through p53- and p21-independent pathways.

E5a of HPV-11 is a transforming oncogene. Previously, we have shown that E5a constitutively activates the expression of protooncogene c-jun by transcriptional regulation through the AP-1 binding site in the c-jun promoter. In the present study, we used two different types of cells: the E5a transfected NIH 3T3 cells and human epidermal keratinocytes, and selectively inhibited different signal transduction pathways to investigate effects of E5a on c-jun expression. We find that protein kinase C and ras-dependent pathways are important for the c-jun induction by E5a, but not the cAMP-dependent pathway.

Previously, we have shown that simian virus 40 (SV40) T antigen can directly cause apoptosis in immortalized human epithelial cells under normal growth conditions. In this study, we further characterized the mechanism of T-antigen-mediated apoptosis involving p53 and whether T antigen can suppress erbB2/neu gene expression. Our results show the differential regulation of erbB2/neu gene expression in different cell clones in response to T antigen transgene, indicating that the regression of erbB2/neu gene by SV40 T is cell-type dependent. Our previous study reported T-antigen-induced apoptosis in p53 mutant cells; however, in this study we find increased levels of p53 protein in T-antigen-containing cells. Therefore, we examined the transactivation function of p53 in these cells. Our data show the failure to transactivate p53, suggesting that increased p53 protein in T antigen expressing cells is functionless at least for transcriptional activation.

Previously, we had reviewed 43 cases of invasive cancers, adenosquamous cell carcinoma and adenocarcinoma for HPV type infections. With the same cases we extended the investigation to cytomegalovirus (CMV) and herpes simplex virus (HSV) infections. Results show that the prevalence of CMV and HSV infections from these cases of cervical carcinoma was 67 and 76%, respectively, by polymerase chain reaction. The results of the analysis of the association of HPV, CMV and HSV with various clinical characteristics of cervical cancer patients indicated that the correlation between HSV infections and clinical stages of squamous carcinoma was marginally significant (P = 0.068). HSV infections seemed to have a higher association with cell keratinization pattern as compared with the other two viral infections.

All-trans-retinoic acid (RA) affects cell growth and regulates gene expression. We examined the expression of topoisomerase 11 gene in Hep3B cells treated with RA. At low RA concentration which did not significantly affect the growth rate of Hep3B cells, RA inhibited the synthesis of topoisomerase II mRNA as revealed by Northern analysis and nuclear run-on analysis. These results indicated that the repression of topoisomerase II gene expression could be directly induced by RA rather than was a secondary event which occurred after cell growth was inhibited by RA. An unexpected finding is that after up to 72 h continuous exposure to RA, the topoisomerase II protein concentration remained unchanged.

E5a of HPV-11 is a transforming oncogene. Previously, we had shown that the E5a gene is required only for the initiation of transformation; c-jun might be involved in the maintenance of transformation. In this study, we exposed E5a transformed NIH 3T3 cells to antisense oligodeoxynucleotides complementary to the 24 nucleotides corresponding to the translation initiation site of the c-jun gene, and examined the effects of this treatment on cell proliferation. Results show that antisense c-jun oligodeoxynucleotides could repress c-jun production and inhibit cell proliferation in E5a transformed cells.