Efeitos da Quercetina sobre a viabilidade e morte celular e expressão gênica de marcadores do ciclo celular em células de Melanoma cutâneo murino B16F10

Abstract

Introduction: Cutaneous melanoma develops from melanocytes, the cells responsible for skin pigmentation, considered a type of cancer that is clinically aggressive due to the precociousness of its development of metastasis. Quercetin (Querc) is a naturally occurring polyphenol that acts as a negative modulator of molecular pathways related to the progression of malignances, including those responsible for cell proliferation and apoptosis. Aim: In the present study, we investigated the effects of Querc on cell viability, death and expression of markers of cell cycle of murine cutaneous Melanoma B16F10 cells. Methods: In our study, B16F10 cells in a suspension of 2 × 104 cells were cultured on the presence of Querc at doses of 200, 100, 50, 25, 12.5 and 6.25 µM for a time period of 48 hours. At the end of this period, we analyzed cell viability using the MTT colorimetric assay (3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromidefor colorimetric assay), the cell proliferation by the expression of mRNA of the A2, B1, D3 and E1 cyclins, using qPCR and colorimetric fluorescence assay with ethidium bromide and orange acridine (BE / AA) for establishing cell death rates. As a control, B16F10 cells cultured in the absence of Querc were subjected to the same analyzes. Results: Our results showed significant differences in the cell viability of B16F10 cells exposed to Querc, highlighting the concentration of 200 µM as the most effective one, since the drug was able of decreasing cell viability by more than 50% at its highest concentration, increasing the events of cell death over 50% as well. Regarding the molecular events that regulate the cell cycle, Querc at the concentration of 200 µM was able of increasing the expression of the cyclin D3, which acts as a blocker of the cell cycle transition from phase G1 to S1, leading, consequently, to a increase on the frequency of B16F10 cells in the G0 / G1 phases of the cell cycle. All of these molecular results were consistent with the changes in the cyclines expression observed in B16F10 cells when treated with Querc at its highest concentration. Conclusions: Our results showed that the treatment with Querc acted as an inductor of cell death, a modulator of cell cycle markers expression and promoter of cell death in B16F10 cells. These findings demonstrate the potential use of Querc as a nutraceutical with adjunctive anti-cancer therapeutic properties.