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Investigation of a new oxazolidine derivative in human resistance acute leukemia cells: deciphering its mechanism of action by label-free proteomic

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Abstract

The present study aimed to evaluate the mechanism of action of the antineoplastic activity of an oxazolidine derivative, LPSF/NB-3 (5-(4-cloro-benzilideno)-3-etil-2-tioxo-oxazolidin-4-ona). Cytotoxicity assays were performed in peripheral blood mononuclear cells (PBMCs) and resistant acute leukemia cell line (HL-60/MX1) by the MTT method. LPSF/NB-3 exhibited cytotoxicity in HL-60/MX1, but it was not toxic to healthy cells in the highest dose tested (100 μM). The protein extract of HL-60/MX1 cells treated with LPSF/NB-3 was subjected to proteomic analysis using two-dimensional chromatography coupled to mass spectrometry. We could identify a total of 2652 proteins, in which 633 were statistically modulated. Within the group of protein considered for the quantitative analysis with the established criteria, 262 were differentially expressed, 146 with increased expression and 116 with decreased expression in the sample treated with LPSF/NB-3 compared to the control. The following differentially expressed pathways were found: involving regulation of the cytoskeleton, DNA damage, and transduce cellular signals. Networks that were highlighted are related to the immune system. The ELISA technique was used to assess the immunomodulatory potential of LPSF/NB-3 in PBMCs. We observed significant decrease of IFNγ (p < 0.01) and dose-response pattern of the cytokines IL-6, IL-17A, IL-22, and IL-10. Therefore, results suggest that LPSF/NB-3 appears to modulate important pathways, including cell cycle and immune system regulatory pathways.

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Acknowledgments

We would like to thank the Brazilian National Research Council (CNPq), the Research Foundation of Pernambuco State (FACEPE), National Institute for Science and Technology in Pharmaceutical Innovation (INCT_if), and CAPES for student fellowships.

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Authors and Affiliations

  1. Research Center for Therapeutic Innovation (NUPIT-SG), Federal University of Pernambuco, Professor Moraes Rêgo s/n, Cidade Universitária, Recife, Pernambuco, 50670-901, Brazil

    Lidiane Vasconcelos do Nascimento Carvalho, Wanessa Layssa Batista de Sena, Michelly Cristiny Pereira, Maira Galdino da Rocha Pitta, Ivan da Rocha Pitta & Moacyr Jesus Barreto de Melo Rêgo

  2. Bone Marrow Transplant Center, National Institute of Cancer, INCA, Rio de Janeiro, Brazil

    Eliana Abdelhay

  3. Drug Planning and Synthesis Laboratory, Federal University of Pernambuco, Recife, Brazil

    Maria do Carmo Alves de Lima

  4. Department of Mass Spectrometry Research and Development, SpectraMass, Campinas, São Paulo, Brazil

    Gustavo Henrique Martins Ferreira Souza

  5. Molecular Biology and Proteomics of Blood Laboratory, LBCD-LADETEC, Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    Luciana Pizzatti

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  1. Lidiane Vasconcelos do Nascimento Carvalho
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Contributions

LVNC and WLBS conducted the bench experiments and made the writing of the manuscript. MGRP, EA, and MCAL provided the necessary reagents for carrying out the experiments and guided the progress of the study. MCP and LP contributed to in silico data analysis. IRP and MJBMR were responsible for study design and data interpretation. All authors read and approved the manuscript and all data were generated in-house and that no paper mill was used.

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Correspondence to Moacyr Jesus Barreto de Melo Rêgo.

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All volunteers signed a consent form and the study was approved by the Human Research Ethics Committee from the Federal University of Pernambuco (CEP/CCS/UFPE No. 11006/12).

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do Nascimento Carvalho, L.V., de Sena, W.L.B., Abdelhay, E. et al. Investigation of a new oxazolidine derivative in human resistance acute leukemia cells: deciphering its mechanism of action by label-free proteomic. Naunyn-Schmiedeberg's Arch Pharmacol 394, 1153–1166 (2021). https://doi.org/10.1007/s00210-020-02024-8

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