Determinación del Impacto del Crecimiento de Cédulas de Cáncer de mama en cultivos en 3D organotípos sobre el perfil de metilación global del ADN y su efecto en la invasión y metástasis.

Abstract

RESUMEN El cáncer de mama es la neoplasia más común en mujeres y la principal causa de muerte por cáncer en México desde 2006. Su desarrollo se ve influenciado por alteraciones genéticas y epigenéticas. Se propone el uso de cultivos en 3D organotípicos para simular mejor el microambiente tumoral en comparación con los cultivos 2D. Este estudio analiza el efecto del cultivo 3D en el perfil de metilación del ADN de diferentes líneas celulares de cáncer de mama, identificando genes metilados asociados a la invasión y metástasis.

ABSTRACT “Breast cancer is the most frequent neoplasia among the female population in the world; in Mexico since 2006, it is the leading cause of cancer death in women. The constant accumulation of genetic and epigenetic alterations in cells influence the development and progression of breast cancer and these changes vary according to the molecular subtype and culture methods. In general, monolayer culture (2D) is the most widely used for the study of cancer biology, however, it presents certain limitations to faithfully recreate the microenvironment and molecular processes of tumors in vivo, therefore, it is has proposed the use of 3D organotypic culture methods that allow optimizing culture conditions to recreate biological and molecular characteristics such as: cell-cell and cell-extracellular matrix interactions, cell heterogeneity, differential growth kinetics, development of different gradients in the microenvironment, so that cells receive stimuli from the local environment resulting in a change in the activity of signaling pathways, gene expression and alteration of the epigenetic pattern that includes DNA methylation. DNA methylation is an epigenetic modification that is widely influenced by the tumor microenvironment and is frequently altered in cancer. In this study, we analyzed the impact of 3D culture of breast cancer cells (Hs578T, BT-474 and SK-BR-3) on the global DNA methylation profile using methylation microarrays, in addition, using a in vivo model of Xenotransplants of breast cancer cells that allowed us to evaluate the phenotypes described in the literature and the invasive and metastatic capacity of the study cell lines that correspond to different molecular subtypes, to classify them according to their aggressiveness and distinguish their invasive capacity and metastatic in vivo. The results obtained allowed us to identify the differentially methylated genes at the promoter level between our cell lines, to group them into shared and exclusive genes of each study cell line, with the above a genetic signature was associated with invasion and metastasis for the three cell lines. study, in addition, a gene was suggested as a potential target for epigenetic therapy against breast cancer due to its molecular characteristics and potential interactions with other proteins”.