Title:Proapoptotic Effects of triazol-1,4-Naphthoquinones Involve Intracellular ROS Production and MAPK/ERK Pathway in Human Leukemia Cells
VOLUME: 20 ISSUE: 17
Author(s):Tangbadioa H. Coulidiati, Bruna B. Dantas, Glaucia V. Faheina-Martins, Enéas Ricardo de Morais Gomes, Juan C.R. Gonçalves* and Demetrius A. Machado de Araújo
Affiliation:Laboratory of Cellular and Molecular Biotechnology, Department of Biotechnology, Federal University of Paraiba, 58051-900, Joao Pessoa-PB, Laboratory of Cellular and Molecular Biotechnology, Department of Biotechnology, Federal University of Paraiba, 58051-900, Joao Pessoa-PB, Laboratory of Cellular and Molecular Biotechnology, Department of Biotechnology, Federal University of Paraiba, 58051-900, Joao Pessoa-PB, Laboratory of Cellular and Molecular Biotechnology, Department of Biotechnology, Federal University of Paraiba, 58051-900, Joao Pessoa-PB, Laboratory of Cellular and Molecular Biotechnology, Department of Biotechnology, Federal University of Paraiba, 58051-900, Joao Pessoa-PB, Laboratory of Cellular and Molecular Biotechnology, Department of Biotechnology, Federal University of Paraiba, 58051-900, Joao Pessoa-PB
Keywords:Cancer, acute leukemia, naphthoquinone, cytotoxicity, HL-60, reactive oxygen species.
Abstract:
Background: The natural products constitute an important source of antitumor and cytotoxic agents.
Naphthoquinones are effectively quinones present in different plants, with demonstrated anticancer activities. A
recent study conducted by our group demonstrated the antileukemic potential of two novel triazol-1,4-
naphthoquinones derivatives, PTN (2-(4-Phenyl-1H-1,2,3-triazol-1-yl)-1,4-naphthoquinone) and MPTN (2-[4-
(4-Methoxyphenyl)-1H-1,2,3-triazol-1-yl]-1,4-naphthoquinone). Although, the mechanisms underlying the
proapoptotic effects of PTN and MPTN have not been fully elucidated so far.
Objective: The aim of this study was to evaluate the proapoptotic mechanism of PTN and MPTN in human
acute leukemia cells.
Methods: We used fluorescence microscopy to observe acridine orange and annexin V staining cells. Flow
cytometry assay has also been used for ROS quantification, BAX and cytochrome c proteins expression and
apoptosis analysis. MTT assay and western blotting technique have been performed as well for MAPK pathway
analysis.
Results: By using the acridine orange and annexin V staining with fluorescence microscopy, we have characterized
the proapoptotic effects of PTN and MPTN in HL-60 cells involving the intrinsic mitochondrial pathway,
since these compounds promoted an increase in the intracellular BAX and cytochrome c protein levels (p<0.05).
We further demonstrated that apoptosis induction in HL-60 cells was mediated by increasing intracellular ROS
levels via ERK but not p38 MAPKs pathway.
Conclusion: Taken together, these results have demonstrated that PTN and MPTN are promising tools for the
development of new anti-leukemic drugs.
