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MELO, V. A. M. R. Construction of tools for study of the possible interaction between interferon-beta and p53. Master thesis – Instituto de Ciencias Biomedicas, Universidade de Sao Paulo, Sao Paulo, 2008. The formation of tumors is related to a combination of factors, including alteration in the control of cellular proliferation, alteration in response to extra cellular signals and tolerance of the tumor’s presence by the immune system. The p53 pathway represents a fundamental axis for the control of proliferation and induction of apoptosis. Interferon-beta (IFNβ) plays an important role in the modulation of immune response, participates in the creation of anti-tumor immunologic memory and also impacts the proliferative and apoptotic potential of tumor cells. Recently, data from the literature has shown that IFNβ activates transcription of p53 and that some components of IFN system exert their function through the p53/p14arf pathway. In this project, we constructed a series of tools to be used to explore the interactions between p53 and IFNβ in a melanoma model. The first tool, a derivative of the B16 cell line with reduced p53 expression due to a specific miRNA, will be useful for the measurement of the IFNβ response in presence or absence of cellular p53 protein. This project also includes the confirmation of plasmid and adenoviral vectors that carry the cDNAs of eGFP, Luciferase, p53 or murine IFNβ. These vectors can be used for the introduction of these factors, alone or in combination, in target cells for evaluation of their impact on gene expression, proliferation or cell death. In preliminary assays, plasmids encoding the Luciferase cDNA with expression directed by a p53-responsive promoter or a promoter responsive to components of IFN system were co-transfected in B16 cells with the p53 and IFNβ expression vectors. Even though the activity of either p53 or IFNβ alone was confirmed, an additive effect of these factors was not observed using this assay. Future studies of the possible interactions between the p53 and IFNβ pathways will benefit from the tools constructed in this project.

Construção de ferramentas para estudo da possível interação entre interferon-beta e p53.