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Combination of dasatinib and okadaic acid induces apoptosis and cell cycle arrest by targeting protein phosphatase PP2A in chronic myeloid leukemia cells

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Abstract

Chronic myeloid leukemia (CML) is a cancer type of the white blood cells and because of BCR-ABL translocation it results in increased tyrosine kinase activity. For this purpose, dasatinib is the second-generation tyrosine kinase inhibitor that is used for inhibition of BCR-ABL. Effectively and safetly, dasatinib has been used for imatinib-intolerant/resistant CML patients. Protein phosphatase 2A (PP2A) is the major serine/threonine phosphatase ensuring cellular homeostasis in cells and is associated with many cancer types including leukemias. In this study, we aimed to investigate the effects of dasatinib and okadaic acid (OA), either alone or in combination, on apoptosis and cell cycle arrest and dasatinib effect on enzyme activity and protein-level changes of PP2A in K562 cell line. The cytotoxic effects of dasatinib were evaluated by WST-1 analysis. Apoptosis was determined by Annexin V and Apo-Direct assays by flow cytometry. Cell cycle arrest analysis was performed for the investigation of the cytostatic effect. We also used OA as a PP2A inhibitor to assess apoptosis and cell cycle arrest changes in case of reducing the level of PP2A. PP2A enyzme activity and protein levels of PP2A were examined by serine/threonine phosphatase assay and Western blot analysis, respectively. Apoptosis was increased with dasatinib and OA combination. Cell cycle arrest was determined especially after OA treatment. The enzyme activity was decreased depending on time after dasatinib application. PP2A regulatory and catalytic subunit protein levels were decreased compared to control. Targeting the PP2A by dasatinib and OA has potential for CML treatment.

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Abbreviations

CML:

Chronic myeloid leukemia

Das:

Dasatinib

OA:

Okadaic acid

TKI:

Tyrosine kinase inhibitor

WST-1:

Water soluble tetrazolium-1

CP:

Chronic phase

AP:

Accelerated phase

IC50 :

Inhibitory concentration 50%

FBS:

Fetal bovine serum

DMSO:

Dimethyl sulfoxide

FITC:

Fluorescein isothiocyanate

PI:

Propidium iodide

PBS:

Phosphate-buffered saline

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Acknowledgements

None.

Funding

This study was financially supported by Ege University Scientific Research Projects Fund (BAP) which is Grant Number 2015-TIP-007.

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Authors and Affiliations

  1. Medical Biology Department, Faculty of Medicine, Ege University, Izmir, Turkey

    Buket Ozel, Sezgi Kipcak, Cigir Biray Avci, Cumhur Gunduz & Nur Selvi Gunel

  2. Division of Haematology, Department of Internal Medicine, Faculty of Medicine, Ege University, Izmir, Turkey

    Guray Saydam

  3. Medical Biology Department, Faculty of Medicine, Beykent University, Istanbul, Turkey

    Cagdas Aktan

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  1. Buket Ozel
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Contributions

Conceptualization; BO, NSG, and GS. Material preparation, data collection, and investigation; BO, SK, and CA. Data analysis and interpretation; BO, CBA, and CG. Writing the manuscript; BO. All authors read and approved the final manuscript.

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Correspondence to Buket Ozel.

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Ozel, B., Kipcak, S., Biray Avci, C. et al. Combination of dasatinib and okadaic acid induces apoptosis and cell cycle arrest by targeting protein phosphatase PP2A in chronic myeloid leukemia cells. Med Oncol 39, 46 (2022). https://doi.org/10.1007/s12032-021-01643-2

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