Vesículas extracelulares derivadas de tecido adiposo como moduladoras do perfil metabólico de células MDAMB- 231: papel das espécies reativas de oxigênio

Abstract

The global scenario of obesity is a growing public health concern due to the multiple clinical and metabolic complications associated with this condition. Obesity is characterized by excess adipose tissue, which contributes significantly to chronic low-grade inflammation that can promote tumor development. Two factors that have been gaining prominence in the relationship between obesity and cancer are oxidative stress and cellular communication mediated by extracellular vesicles. In obesity, the release of these vesicles is increased, intensifying the transport of molecular signals that modulate the tumor microenvironment. Oxidative stress results from energy imbalance combined with the reduction in the antioxidant capacity of adipose tissue. It is closely linked to tumor progression and worse cancer prognosis in obese patients. The present study aimed to evaluate the production of intracellular reactive oxygen species (ROS) and their effects on cells of the MDA-MB-231 breast tumor cell line stimulated with vesicles derived from adipose tissue of obese and eutrophic individuals, the latter being the control group. For this purpose, the mitochondrial ROS synthesis inhibitor MitoTEMPO was used, and ROS production was evaluated through two probes: CM-H2DCFDA and MITOSOX. After analyzing ROS production, the cell migration process was assessed through the wound healing assay, and the MitoTEMPO inhibitor was used to evaluate the relationship between oxidative stress and cellular processes capable of favoring tumor development and progression. Finally, we assessed the global metabolic profile of the cells using the metabolomics technique with a non-targeted approach through LC/MS. For this experiment, the cells were stimulated with vesicles from eutrophic and obese patients in the presence and absence of MitoTEMPO, seeking to qualitatively and quantitatively evaluate the most relevant metabolites between the groups. Our results demonstrated that vesicles derived from obese adipose tissue increase mitochondrial ROS production in MDA-MB-231 cells, and this effect is suppressed in the presence of the inhibitor MitoTEMPO. Furthermore, an increase in the migration of cells stimulated by vesicles from obese adipose tissue was observed compared to the control group, suggesting that ROS mediates this migration. Together, these results indicate that vesicles derived from obese individuals increase ROS production in MDAMB-231 cells, which are related to changes in their function and metabolism, participating in the modulation of mechanisms related to tumor progression.