ABSTRACT
OBJECTIVE: The goal of this study was to study whether estrogen could induce progression of cervical neoplasia by the influence of direct hormonal transactivation of the viral genes. METHODS: We examined the in vivo effect of estrogen on HPV-18 URR E6/E7 transgenic mice. We analyzed the growth stimulation of epithelial cells at squamo-columnar junction and vagina and the expression of HPV E6/E7 in transgenic mice. The promoter activity of HPV-18 URR after treatment of estrogen was also evaluated by in vitro transient transfection assay. RESULTS: The dysplastic lesions of lower genital tract were more frequently seen in the HPV-18 URR E6/E7 transgenic mice and estrogen-treated mice, when compared to those of control group (P < 0.05). The majority of cells in whole epithelial layer of cervix and vagina were proliferating cell nuclear antigen-positive (PCNA) by immunohistochemical staining in the estrogen-treated transgenic mice. Hyperplastic glandular lesions were also identified in estrogen-treated HPV-18 URR E6/E7 transgenic mice (5 of 21) and estrogen-treated nontransgenic mice (2 of 10). The level of E6/E7 transcripts in transgenic mouse was increased in the presence of estradiol. Treatment with 0.5 x 10(-6) M estradiol in the presence of cotransfection with the estrogen receptor expression vector and URR-CAT showed an additive effect of CAT activity (4.8-fold increase). CONCLUSION: The HPV E6 and E7 oncogenes implicated in cervical cancer are indeed capable of potentiating tumor formation in animal model. Continual estrogen-induced proliferation might be viewed by in vivo and in vitro mechanisms by which squamous cells as well as glandular cells in cervix and vagina became permissive for neoplastic progression in HPV-18 URR E6/E7 transgenic mice and their molecular systems.
Subject(s)
Cell Transformation, Viral/drug effects , DNA-Binding Proteins , Estradiol/toxicity , Oncogene Proteins, Viral/genetics , Papillomaviridae/genetics , Uterine Cervical Neoplasms/etiology , Vaginal Neoplasms/etiology , Animals , Epithelium/drug effects , Epithelium/virology , Female , Gene Expression Regulation, Viral/drug effects , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Transgenic , Regulatory Sequences, Nucleic Acid/drug effects , Regulatory Sequences, Nucleic Acid/genetics , Transcriptional Activation/drug effects , Up-Regulation/drug effects , Uterine Cervical Neoplasms/chemically induced , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/virology , Vaginal Neoplasms/chemically induced , Vaginal Neoplasms/pathology , Vaginal Neoplasms/virologyABSTRACT
We have previously reported that human papillomavirus (HPV) E7 interacts with IRF-1, a key regulator of cellular immune response, and abrogates its transactivation function at the molecular level in vitro. To confirm our previous data, we investigated in vivo the E7-mediated down-regulation of IRF-1 using HPV E7-inducible cells and transgenic mice expressing HPV-18 E6/E7. When E7 was induced in the absence of tetracycline, the expression of target genes of IRF-1 (TAP-1, IFN-beta, MCP-1 that are important for cellular immunity) was clearly reduced as determined by RT-PCR. In addition, the IRF-1 activity was down-regulated in E7-expressing cells into which IFN-beta-CAT reporter plasmid was transfected. To further investigate the E7-mediated immune regulation in vivo, we constructed transgenic mice expressing E6 and E7 genes of HPV-18 under the control of HPV-18 promoter (URR). From several rounds of breeding, we obtained from a transgenic line that developed a cervical dysplasia and expressed E6/E7 as determined by histological examination and RT-PCR, respectively. Subsequent RT-PCR analysis indicated that TAP-1, IFN-beta, and MCP-1 genes were less expressed in a cervical tissue derived from transgenic mouse, when compared with a cervix derived from normal mouse. From these results, we conclude that the E7 transgene expression inactivates the transactivation function of IRF-1 in vivo, which might be important for the elucidation of the E7-mediated immune evading mechanism that is frequently found in cervical cancer.
