Avaliação in vitro e in silico da atividade do S-(-)-Álcool Perílico sobre linhagens celulares do Carcinoma Epidermóide de Língua

Abstract

Oral squamous cell carcinoma (OSCC) is the most frequent malignant neoplasm of the oral cavity and constitutes a public health problem due to its high incidence and mortality rate caused in many cases by therapeutic failure and tumor resistance. Therefore, the search for new biologically active molecules stands out, such as those found in products of natural origin. This work aims to evaluate the antineoplastic activity of S-(-)-perillyl alcohol (POH) in cell cultures of tongue CEO and to predict its affinity through a computer model on proteins that regulate the cell cycle. For this purpose, two cell lines of tongue CEO were used, HSC-3 and SCC-25. The following groups were analyzed: G0 (control; cells cultured in the absence of POH), G1 (cells treated with 40 μM cisplatin), G2 (cells treated with 0.5 mM POH), G3 (cells treated with 1 .0 mM), G4 (cells treated with 1.5 mM POH) and G5 (cells treated with 3.0 mM POH). Differences between these groups were investigated through the following assays: cell viability (Alamar Blue and Live/Dead assay) and migratory activity (Wound healing). Affinity prediction between POH and cell cycle control molecules were also performed using molecular docking using Molegro Virtual Docker, v. 6.0.1. The data was statistically treated by GraphPad Prism 6.0 (GraphPad Software, USA), parametric analysis using Anova test, Tukey post-test and Kruskal-Wallis non-parametric statistical test, followed by t student test were adopted for determination of differences between the experimental groups. The significance index considered in this work was 5%. For both cell viability assessment techniques (Alamar Blue and Live/dead assay) analyzed in this work, POH was able to reduce the cell viability of tongue CEO lines in a dose-dependent and time-dependent manner (p<0 .05). The concentrations of 1.5 mM and 3 mM of POH obtained better or similar results to those found in 40 μM cisplatin, for the two strains, in the evaluation of cell viability (p<0.05). The IC50 values of POH were 1.5 mM for the SCC-25 cell at all time intervals (24 h, 48 h and 72 h), since for the HSC-3 line they were 3 mM for 24 h and 48 h times and 1.5 mM for the 72 h interval. POH was able to inhibit the migration of the two DSC cell lines in a concentration-dependent manner (p≤0.05), compared to the control group. The ability of the POH molecule to bind to proteins responsible for cell cycle activation was evaluated using docking models. Among them, the protein GTPase Kras showed the best binding energy (-86.70 kcal/mol), featuring hydrogen bonds with residues THR58 (A) and ASP57 (A) and steric bonds with residues TRY32 (A) and ALA18 ( THE). The evidence from this study supports the idea that POH has antineoplastic activity on the CEO, suggesting that this molecule may be a strong candidate for the development of drugs aimed at the treatment of this pathology.