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Characterization of a multidrug-resistant chronic myeloid leukemia cell line presenting multiple resistance mechanisms

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Abstract

The multidrug-resistant (MDR) phenotype is multifactorial, and cell lines presenting multiple resistance mechanisms might be good models to understand the importance of the various pathways involved. The present work characterized a MDR chronic myeloid leukemia cell line, derived from K562 through a selective process using daunorubicin. This MDR cell line was shown to be resistant to vincristine, daunorubicin, and partially resistant to imatinib. It showed a slower duplication rate. Overexpression of ABCB1 and ABCC1 was observed at the protein and functional levels and the expression of CD95, a molecule related to cell death, was reduced in the MDR cell line. Conversely, no differences were observed related to the anti-apoptotic molecule Bcl-2 or p53 expression. The activation antigen CD69 was reduced in the MDR cell line and treatment with imatinib further decreased the expressed levels. Furthermore, secretion of IL-8 was diminished in the MDR cell line. When daunorubicin-selected cells were compared to another MDR cell line, Lucena 1, derived from the same parental line K562, and selected with vincristine, a different profile was observed in relation to most aspects studied. When both cell lines were silenced for ABCB1, differences in CD69 and CD95 were maintained, despite resistance reversal. These results reinforce the idea that cell lines selected in vitro may display multiple resistance strategies that may vary with the selective agent used as well as during different steps of the selection process.

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Abbreviations

CML:

Chronic myeloid leukemia

ABC transporters:

ATP binding cassette transporters

P gp:

P glycoprotein

BCRP:

Breast cancer resistant protein

MDR:

Multidrug resistance

MRP1:

Multidrug resistance related protein 1

IM:

Imatinibe mesylate

FCS:

Fetal calf serum

VCR:

Vincristine sulfate

DNR:

Daunorubicin hydrochloride

CDDP:

Cisplatin

VP:

Verapamil

DMSO:

Dimethyl sulfoxide

CFDA:

Carboxy fluorescein diacetate

Rho:

Rhodamine 123

INDO:

Indomethacin

PRB:

Probenicid

FACS:

Fluorescence cell sorter

PBS:

Phosphate buffered saline

HBSS:

Hank’s balanced salt solution

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Acknowledgments

We thank Thais M. Gameiro Marques and Bruno Paredes for their assistance in performing the IL-8 measurements. We also thank the Brazilian agencies: Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq, Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro—FAPERJ and FAPERJ/PP SUS, and INCT-Controle do Cancer for financial support.

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Authors and Affiliations

  1. Laboratório de Imunologia Tumoral, Instituto de Bioquímica Médica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Bloco H02, Sala 003, Rio de Janeiro, 21941-590, Brazil

    Nathalia Daflon-Yunes, Flavio Eduardo Pinto-Silva, Raphael Silveira Vidal, Bruna Fortunato Novis, Tandressa Berguetti & Vivian M. Rumjanek

  2. Instituto de Biofísica Carlos Chagas Filho, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Bloco G01, Rio de Janeiro, 21941-590, Brazil

    Raphael Silveira Vidal

  3. Laboratório de Artropodos e Hematofogos, Instituto de Bioquímica Médica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Bloco Dss, Sala 05, Rio de Janeiro, 21941-590, Brazil

    Raphael Rodrigues Soares Lopes & Carla Polycarpo

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  1. Nathalia Daflon-Yunes
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  2. Flavio Eduardo Pinto-Silva
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  3. Raphael Silveira Vidal
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  4. Bruna Fortunato Novis
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  6. Raphael Rodrigues Soares Lopes
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  8. Vivian M. Rumjanek
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Correspondence to Vivian M. Rumjanek.

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Nathalia Daflon-Yunes and Flavio Eduardo Pinto-Silva have contributed equally to this work.

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Daflon-Yunes, N., Pinto-Silva, F.E., Vidal, R.S. et al. Characterization of a multidrug-resistant chronic myeloid leukemia cell line presenting multiple resistance mechanisms. Mol Cell Biochem 383, 123–135 (2013). https://doi.org/10.1007/s11010-013-1761-0

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