Uso de Herramientas Bioinformáticas para el análisis de la regulación epigenética de la expresión de proteínas involucradas en procesos tumorales

BUGNON VALDANO, MP; CAVATORTA, AL; MARZIALI, F; FACCIUTO, F; BOCCARDO, E; GARDIOL, D

Zavalla

Congreso; XV Congreso y XXXIII Reunión Anual de la Sociedad de Biología de Rosario; 2013

Sociedad de Biología de Rosario

Malignant transformation is associated with loss of cell polarity. We are focused on the study of polarity proteins, such as the human homologous of the Drosophila tumour suppressor, Disc large 1 (DLG1). DLG1 locates at cell-cell contacts, where it has both, structural and signalling roles. It has been proposed to have tumour suppressor functions: it inhibits epithelial cell proliferation; it is a target of several viral oncoproteins and its expression is markedly reduced or absent in the late stages of different tumours. However, DLG1 has also been proposed to have oncogenic functions: mainly, DLG1 is increased and delocalized in the intermediate stages of the malignant progression. We want to characterize the regulation of DLG1 in order to understand its function in carcinogenic processes. Altered patterns of DNA methylation are key mechanisms in tumour progression. To study whether epigenetic regulation could explain the unusual pattern of DLG1 in biopsies, we focus on the analysis of the methylation status of its promoter. Using several bioinformatics tools, we analysed the promoter and established the presence of a region with a high density of CpG dinucleotides, liable to be regulated by methylation. We applied the method of treatment of genomic DNA with sodium bisulfite, subsequent PCR and sequencing, establishing reliably the status of each CpG. We designed BSP primers, suitable for amplification of bisulfite-treated DNA. The goal of this study is to establish if there is a correlation between promoter methylation status and DLG1 expression.