Avaliação do efeito antiproliferativo de bis-chalconas sintéticas (L05 E L08) em linhagens tumorais e não tumorais

Abstract

Cancer is characterized by a set of different types of malignant alterations, of genetic and epigenetic origin, which culminate in the disordered growth of abnormal cells. The most advanced pharmacological therapies currently involve the administration of chemotherapy and non-specific hormone-blocking agents, which lead to various side effects and impair adherence to treatment. For this reason, research into new synthetic molecules, such as the bis-chalcones L05 and L08, is promising, since there have been studies showing various anticancer effects of substances belonging to the same class. This study aimed to investigate the antiproliferative profile of the synthetic bis-chalcones L05 and L08 through in vitro experiments. The tests used to assess the cytotoxicity of the bis-chalcones against various tumor and non-tumor cell lines were: the MTT method, at 24, 48 and 72 hours, with treatment concentration decreasing by a ratio of 2, starting at 100 µM; the time-concentration pulse test, at 24 and 30 hours, also starting at a concentration of 100 µM; the MTT assay with SDS, at 72 hours, starting at a treatment concentration of 25 µM. In addition, the cell viability test was carried out using Trypan blue at 24 and 48 hours, at treatment concentrations of 2, 4, 8 and 16 µM, and finally, the morphological test using rapid panopticon, at the same concentrations and treatment times as the previous test. Using the MTT method, L05 showed an effect against four cancerous lineages, with the lowest IC50 values being 10.7 µM and 16.8 µM in Mel-501 and HCT-116, respectively, while showing no cytotoxicity for the non-tumorous lineage. It can be seen that L08 had a better effect on the cell lines tested, since it was active in a greater number of lines, with the lowest IC50 values being 2.5 µM and 4.7 µM in HCT-116 and Mel-501, respectively. In view of these results found in the 72-hour MTT, it was decided to continue the research using only the unpublished bis chalcone L08 and the HCT-116 strain. With regard to the cell viability test, it was observed that L08 was able to significantly reduce the number of viable cells, from the lowest concentration used for both treatment times, without modifying the number of non-viable cells with statistical significance. Morphological analysis showed the appearance of changes suggestive of the start of a cell death process, such as prolongation of the plasma membrane (“blebs” or apoptotic bodies), the appearance of vacuoles in the cytoplasm, chromatin condensation (pyknosis) and nuclear fragmentation, as well as the appearance of cell debris. Finally, the results of this study show a time-dependent cytotoxic effect of the synthetic and novel bis-chalcones L05 and L08, with the second sample being the most active, which also promoted the onset of cell death. In order to further investigate the mechanisms involved in cytotoxicity and cell death, further in vitro and in vivo experiments are needed to elucidate and ratify their antiproliferative action on HCT-116 and other cell lines.