Estabelecimento de um modelo tridimensional e avaliação do comportamento da 22ß-hidroxitingenona em células de melanoma

Abstract

In the study of cancer, three-dimensional cell culture models combined with investigating pharmacokinetic and cytotoxic properties in silico have become valuable tools for exploring the tumor microenvironment, disease advances and the search for new therapeutic targets. Evaluating the cellular metabolic profile in different tumor microenvironments of melanoma cell lines SKMEL-19, SKMEL-28 and AKMEL-103 against the action of 22-HTG was the objective of this study. Virtual 22-HTG screening showed interaction between 22-HTG and the AKR1B10 gene. Evaluating the generation and characterization of melanoma microtissues, it was observed that the SKMEL-19 and SKMEL-103 cell lines, when cultivated in spheroids, remained more regular on the fifth day of culture, at the 100x104 concentration, and the SKMEL-28 and MRC5 cell lines, when grown in spheroids remains more regular on the fifth day of cultivation, at the 50x104 concentration. To evaluate the metabolic profile, viability, glucose consumption and LDH activity assays were performed under three culture conditions (25, 5.5 and 0.5 mM of glucose). Regarding cell viability, a decrease was observed in SKMEL28 and MRC5 strains when cultured in 0.5 mM glucose. Regarding glucose consumption, all melanoma strains showed significant consumption (p<0.001) when compared to the normal strain in both culture media. In medium with scarce glucose (medium with 0.5 mM), strains MRC-5, SKMEL 28 and SKMEL 103 showed a significant increase in LDH enzymatic activity. According to the analyses, it was verified that the drug 22-HTG leads to a reduction in the enzymatic activity of the enzyme LDH and in the glucose uptake significantly in the SKMEL 28 and SKMEL 103 strains both grown in monolayers and in a three-dimensional model. This is the first study to describe the activity of 22β- hydroxytingenone against human melanoma cells grown in a three-dimensional model, which gives an innovative aspect to this work. The inclusion of the tumor microenvironment in a three-dimensional model using melanoma cells proved to be effective and feasible for the study of 22β-hydroxytingenone, revealing its ability to act differently in the metabolism of these cell lines. This approach is fundamental to better understand the interaction between natural substances and the tumor microenvironment, allowing the development of more effective and targeted therapeutic strategies against melanoma cancer.