Efeito do meio condicionado em células pré-tratadas com extrato hidroalcoólico de guaraná em células tumorais

Abstract

The tumor microenvironment is a determinant in the programming, regression or progression of the tumor, as it has the ability to modulate tumor cells through the release of soluble factors in the environment. The study of the antineoplastic effect of drugs, as well as that of herbal medicines, encompasses the action of different cell types and how the action of these compounds modulate these cells present in the tumor microenvironment. In addition, in the scenario of herbal medicines with antineoplastic action, guarana (Paullinia cupana) has emerged as a candidate. Objective: The objective of this work was to evaluate the direct effect of the hydroalcoholic extract of guarana (EHG) on different cell lines and the effect of conditioned media (MCs) on macrophages and normal epithelial cells pretreated with EHG on the proliferative and migratory profile of breast and lung tumor cells. Materials and Methods: The values of the minimum inhibitory concentration of EHG, as well as the antioxidant activity and evaluation of the death profile were conducted in cell lines. Conditioned media were generated in cells pretreated with the highest non-cytotoxic concentration of EHG. The effects of secretoma were evaluated in tumor cells by means of the clonogenicity test, the assessment of viability, cell migration and analysis of the cell cycle profile. Results: The results showed that the values obtained from direct treatment with EHG were heterogeneous among the

different cell lines evaluated: MDA-MB231, MCF7, MCF10A, A549, BEAS-2B, NIH- 3T3, MRC5, NCTC clone 929, differentiated THP-1 in macrophage. The most

resistant strain was lung adenocarcinoma (IC50=17.652±10.22 μg/ml), while the most sensitive was the murine fibroblast (IC50=1.504±34.78 μg/ml). EHG demonstrated antioxidant activity by reducing the levels of reactive oxygen species (ROS) in the MDA-MB 231 strain, both at baseline and under oxidative stress conditions. Direct treatment of EHG for 24h with the IC50 value (12,022±10.73 μg/ml) directs BEAS-2B cells to an apoptotic death profile. The results showed that the MCs in MCF10A were able to reduce the viability and clonogenic capacity of A549, while those in MCF7 only reduced the clonogenic capacity. The MCF10A MC CTL increased the amount of A549 cell cycle G1 phase cells. MC from EHG-pretreated lung epithelial cells increases the wound closure capacity of A549 cells while reducing clonogenic capacity. The MCF7 lineage when treated with secretoma of differentiated cells into macrophages reduces viability within 72 hours of treatment, but the group that was stimulated with LPS restores cell viability, these data are not visualized in the clonogenic assay where all groups reduce clonogenic capacity. A549 is responsive only to groups that are stimulated with LPS in the clonogenic, increasing the percentage of migration of cells that were treated with secretoma of differentiated cells into macrophages pretreated with EHG and stimulated with LPS, while stimulating the formation of vacuoles in the tumor cells and changes morphology to fusiform, losing cell-cell adhesion. Conclusion: MCs act in different ways in the proliferative capacities of the studied tumor lines, however it is observed that the secretoma of cells pretreated with EHG increased the migratory capacity of A549. Future experiments are needed to elucidate the mechanism of action of the EHG in the modulation of macrophage secretama, as well as to evaluate the induction of the epithelial-mesenchymal transition process in tumor cells treated with the conditioned medium.