Dados do Trabalho

Título

1,4-NAPHTHOQUINONE (CNN1) PROMOTES CELL DEATH IN LEVELS SIMILAR TO IRINOTECAN TROUGH A DRUGGABLE TARGET IN MULTIDRUG RESISTANCE PHENOTYPE OF CHRONIC MYELOID LEUKEMIA CELL LINE

Introdução

Multidrug resistance (MDR) phenotype is well recognized in clinical practice and continues to be one of the major obstacles in Chronic Myeloid Leukemia (CML) treatment. Approximately 20–40% of patients develop resistance at some point during therapy. In this scenario, is important to develop anticancer agent able of circumscribe the multidrug resistance in CML.

Objetivo

Therefore, the aim of this study was to evaluate cytotoxicity and genotoxicity of CNN1 naphthoquinone and SN-38 against CML cell lines.

Métodos

Cytotoxicity analyses of the compounds CNN1 and SN-38 were performed by the Alamar Blue. Mitochondrial membrane potential analysis and membrane integrity were quantified by flow cytometry, followed by cell cycle. Genotoxicity was assessed by using comet assay. Analysis of morphological changes were stained using the Fast Panoptic kit (LaborClin®, PR, Brazil). Molecular docking was evaluated to observe Tyrosyl-DNA Phosphodiesterase 1 (TDP1) interaction with CNN1 using AutoDock Vina program.

Resultados

CNN1displayed similar cytotoxic activity when compared to SN-38 against FEPS cell line with a CI50 of 0.60 µM and CI50 of 0.50 µM, respectively. In membrane integrity test and mitochondrial membrane potential analysis, CNN1 promoted significant disruption of membrane integrity and depolarization of mitochondrial membrane potential more pronounced in FEPS (p< 0.001) cell line than SN-38. Moreover, CNN1 caused cell cycle arrest in G2/M phase in both cell lines (p< 0.001) and SN-38 also caused accumulation of cells in G2/M phase. Alkaline comet assay revealed that CNN1 induced genotoxic effect in K-562 and FEPS (p< 0.001), and did not induce significant DNA damage in PBMC cells. We have found typical hallmark of apoptosis, such phosphatidylserine externalization induced by CNN1 (p< 0.001) in levels similar to SN-38 in both cell lines. Molecular docking simulation predicted an -7,3 kcal/mol binding affinity of CNN1 with TDP1 with π interactions to HIS-263, HIS-493 and ASN-283 from the active site of the enzyme.

Conclusões

Our, results demonstrated that CNN1 to be a promising antitumoral agent against resistant (FEPS) and sensitive (K-562) leukemia cell lines, and revealed cell death in levels similar to irinotecan that can be correlated with pharmacological target for CML therapy.

Palavras-chave

Chronic Myeloid Leukemia; Naphthoquinone; Cytotoxic Activity, genotoxicity; TDP1 protein.

Financiador do resumo

This study was supported by Brazilian funding agencies National Counsel of Technological and Scientific Development (CNPq) and Coordination for the Improvement of Higher Education Personnel (CAPES).