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Molecular and Cellular Biochemistry

, Volume 383, Issue 1–2, pp 123–135 | Cite as

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

  • Nathalia Daflon-Yunes
  • Flavio Eduardo Pinto-Silva
  • Raphael Silveira Vidal
  • Bruna Fortunato Novis
  • Tandressa Berguetti
  • Raphael Rodrigues Soares Lopes
  • Carla Polycarpo
  • Vivian M. RumjanekEmail author
Article

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.

Keywords

P glycoprotein Multidrug resistance Leukemia Cell cycle CD69 

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

Notes

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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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Nathalia Daflon-Yunes
    • 1
  • Flavio Eduardo Pinto-Silva
    • 1
  • Raphael Silveira Vidal
    • 1
    • 2
  • Bruna Fortunato Novis
    • 1
  • Tandressa Berguetti
    • 1
  • Raphael Rodrigues Soares Lopes
    • 3
  • Carla Polycarpo
    • 3
  • Vivian M. Rumjanek
    • 1
    Email author
  1. 1.Laboratório de Imunologia Tumoral, Instituto de Bioquímica Médica, Centro de Ciências da SaúdeUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  2. 2.Instituto de Biofísica Carlos Chagas Filho, Centro de Ciências da SaúdeUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  3. 3.Laboratório de Artropodos e Hematofogos, Instituto de Bioquímica Médica, Centro de Ciências da SaúdeUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil

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