Estabelecimento de modelo de carcinoma in situ para estudos de microambiente tumoral in vitro

Abstract

Three-dimensional in vitro cell culture models are becoming increasingly well-known and applicable in research laboratories due to their ability to mimic tissue-like structures compared to two-dimensional monolayer cell culture models. In cancer research, they have become valuable tools for investigating the tumor microenvironment, cancer stages, and the exploration of new therapeutic targets. In this regard, this study aims to contribute to the validation of an in vitro carcinoma in situ model for the study of the tumor microenvironment and to assess its suitability for anticancer therapy research. Two models were developed using squamous cell carcinoma cell lines: CCE-Skin and CCEinsitu-Skin, with the latter incorporating tumor spheroids into the experimental skin. Morphological studies were conducted to evaluate the structure and presence of cells in the models. For applicability studies, 22-β-hydroxytingenone (22-HTG) was selected based on cytotoxicity and cell invasion aspects. The results indicate that the HaCat-Skin model shows potential for irritation and permeation studies in in vitro reconstructed skin models. The squamous cell carcinoma spheroids exhibited improved morphology after 10 days of culture using micromolds. 22-β-hydroxytingenone demonstrated cytotoxicity in both models with and without tumor cells, and the models allowed for the determination of matrix metalloproteinase proteolytic activity after seven days of culture, as well as through the in situ zymography method. The obtained results are relevant for advancing research in the field of three-dimensional cell culture and may contribute to the development of new methodologies for cancer study and novel therapeutic approaches.