Avaliação dos efeitos de meio condicionado derivado de células de câncer de mama triplo negativo tratadas com ecdisterona sobre fibroblastos: um foco na sinalização purinérgica

Abstract

Triple-negative breast cancer (TNBC) is the most aggressive BC subtype, that presents an enrichment of fibroblasts in its immunosuppressive microenvironment that contributes to tumor cell progression and resistance. The mechanisms involved in this interaction remain poorly understood, especially when natural compounds are used. In the present study, the effects of Ecdysterone on modulating TNBC cell migration and how its conditioned medium alters the behavior of human fibroblasts were investigated, highlighting the role of purinergic signaling in this context. To this end, MDA-MB231 (TNBC) and HFF-1 (fibroblasts) cells were treated with Ecdysterone (20E), and horizontal and vertical migration assays were conducted with non-cytotoxic concentrations. Interestingly, Ecdysterone influenced the migratory behavior of MDA-MB-231 cells, especially vertically, suggesting its potential to contain the invasion of these cells. Furthermore, its conditioned medium inhibited horizontal fibroblast migration, which did not occur when HFF cells were treated directly with the compound. Computational analyses demonstrated the potential of 20E to interact with CD73 and the A2A and A2B adenosine receptors. For CD73, 20E exhibited a higher binding affinity than the endogenous ligand and comparable affinity to synthetic inhibitors. In fibroblasts treated with conditioned medium, ADORA2B (gene encoding A2B) transcript levels were significantly reduced, whereas NT5E (gene encoding CD73) transcript levels were increased. Taken together, the results demonstrate that 20E, even at non-cytotoxic concentrations, alters the behavior of TNBC cells and interferes with their communication with cells within the tumor microenvironment, suggesting the involvement of purinergic signaling and its potential role in modulating tumor–stroma interactions.