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Evaluation of glucocorticoid sensitivity in 697 pre-B acute lymphoblastic leukemia cells after overexpression or silencing of MAP kinase phosphatase-1

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Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

To determine the effect of modulating MAP kinase phosphatase-1 (MKP-1) expression levels on cell death induced by glucocorticoid (GC) or hydroxyurea (HU) treatment in the human pre-B acute lymphoblastic leukemia cell line 697.

Methods

Stable MKP-1 overexpressing transformants of the 697 pre-B acute lymphoblastic leukemia cell line were created and tested for sensitivity to the GC triamcinolone acetonide (TA) and HU, and compared to a control 697 cell line containing normal MKP-1 expression levels. Small interfering RNAs (siRNAs) were designed to inhibit MKP-1 expression and evaluated for their effect on GC-mediated cell death.

Results

MKP-1 overexpression caused a phenotype of partial resistance to HU-induced apoptosis but not to GC-induced apoptosis. Electroporation of siRNAs effectively silenced MKP-1 expression, and increased sensitivity to TA by 9.6±1.9%.

Conclusions

Because MKP-1 protects certain tumor cells from chemotherapy-induced apoptosis, its inhibition is being considered as a possible strategy for combination cancer therapy. However, this study suggests that while MKP-1 inhibition may improve the efficacy of DNA damaging agents, it may have only limited utility in combination with glucocorticoids.

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Abbreviations

GC:

Glucocorticoid

ALL:

Acute lymphoblastic leukemia

GR:

Glucocorticoid receptor

MKP-1:

MAP kinase phosphatase-1

RNAi:

RNA interference

siRNA:

Small-interfering RNA

FITC:

Fluorescein isothiocyanate

μF:

Microfarads

TA:

Triamcinolone acetonide

HU:

Hydroxyurea

RT-PCR:

Reverse transcriptase-polymerase chain reaction

7-AAD:

7-aminoactinomycin D.

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Acknowledgements

The authors would like to thank Paul Hallberg for providing flow cytometry expertise, and Dr. Chris Franklin and Dr. Steve Keyes for providing MKP1 cDNA constructs

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Correspondence to Eric Wickstrom.

Additional information

Grant support: DOE/BER ER63055 to E.W.

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Abrams, M.T., Robertson, N.M., Litwack, G. et al. Evaluation of glucocorticoid sensitivity in 697 pre-B acute lymphoblastic leukemia cells after overexpression or silencing of MAP kinase phosphatase-1. J Cancer Res Clin Oncol 131, 347–354 (2005). https://doi.org/10.1007/s00432-004-0659-3

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  • DOI: https://doi.org/10.1007/s00432-004-0659-3

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