Atividade anticâncer de uma furano-naftoquinona em modelos de cultura celular de melanoma

Abstract

Cancer is a disease characterized by the uncontrolled growth and spread of abnormal cells. Among the main neoplasms, melanoma is the leading disease related to the skin by having high aggressiveness and resistance to conventional chemotherapy in advanced stages of the disease, accounting for 80% of deaths from skin cancer. In search of new therapeutic alternatives, has investigated the use of cell culture models that mimetizem the Tumor Microenvironment in vivo allowing more reliable analysis of in vitro activity of promising drugs for cancer treatment. One of the alternatives is the three-dimensional model of dermal skin structure, also called dermal equivalent. The 1 h-ciclopenta [b] nafto [2.3-d] furan-5.10 (3aH, 10bH)-Dione (IVS320), a furan naphthoquinone synthesized from 2-hydroxy-1.4-naphthoquinone (Lausona), presented excellent results of cytotoxicity against the strains of melanoma justifying the selection of this substance for this research project and the need for the development of other analysis showing their potential. Thus, the present study aimed to standardize the cell culture model in dermal equivalent, as well as assess the anticancer activity of IVS320 in human melanoma line SK-Mel-28. The model of study provided a comparison of the cytotoxic activity is genotoxic activity of Metalloproteinases and of IVS320 in SK-Mel-28 cultivated dermal equivalent culture in monolayer and the ability to inhibit cell migration and cell death mechanism. IVS320 performed for SK-cytotoxic Mel-28 in both cell culture models by the method of the MTT, however its activity was decreased when tested in the dermal equivalent model. CI50 average of IVS320 in SK-Mel-28 grown in monolayer was 0.5 μM in three days of treatment. Assessing the cytotoxicity in dermal equivalent model, introduced IVS320 increase in value from the CI50 concentration and time dependent, 8.05, 5.22 and 2.7 μM in 24, 48 and 72 hours of treatment respectively. The Comet test showed themselves simple tape breaks in three concentrations tested and double tape breaks only in the highest concentration in cells grown in monolayer. In the dermal equivalent model only damage were detected in neutral pH Comet in the two largest concentrations. By zymography assay was possible to assess the activity of Metalloproteinases-2 and-9. The results obtained showed the inhibitory effect of IVS320 enzyme activity in three concentrations tested in both cell culture models in treatment time of 18 hours. Additionally, cell motility test, it was observed that IVS320 also proved effective as the inhibition of this activity in all the three days of treatment (6, 12 and 24 hours) when tested in highest concentration. On differential staining LA/BE, noted an increase in the number of apoptotic cells of concentration dependent manner, however, the number of necrotic cells not varied. The results of this survey provide information about IVS320 and help in studies on alternative means in experimental Pharmacology through use of cell culture models dermal equivalent, and highlights the importance of new experimental models.