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Additive apoptotic effect of STI571 with the cytoprotective agent amifostine in K-562 cell line

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Abstract

Purpose

To study the apoptotic effect of the 2-phenylaminopyrimidine derivative STI571 in combination with antioxidant agents on K-562 cell line derived from a Philadelphia chromosome-positive chronic myeloid leukemia patient.

Materials and methods

K-562 (BCR/ABL+), U-937, and HL60 (BCR/ABL−) leukemic cell lines were incubated with STI571 and the antioxidant agents catalase, glutathione, superoxide dismutase, and amifostine (AMI). Apoptotic effect was analyzed by morphological and flow cytometric criteria.

Results

STI571 at concentrations higher than 0.25 μmol L−1 produced apoptosis (P<0.05) in K-562 cells only after treatment for 72 h. At the mentioned concentrations, STI571 also induced an increase in the loss of mitochondrial transmembrane potential from 24.6 to 40%. Combination of STI571 (0.5 μmol L−1) with antioxidant agents showed that the cytoprotective agent AMI (0.75 mg mL−1) produced an additive effect in the proapoptotic activity of STI571 in K-562 cells at nuclear (58.8%±2.0 vs. 28.9%±3.3) and mitochondrial (53.3%±3.6 vs. 29.5%±1.2) levels.

Conclusions

Our results show that only AMI in combination with STI571, at submicromolar concentration, has an additive effect in K-562 cell line, and it does not have severe toxic effects on Philadelphia chromosome negative cells.

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Abbreviations

AlloBMT:

Allogeneic bone-marrow transplantation

AMI:

Amifostine

ATP:

Adenosine triphosphate

BC:

Fatal blastic phase

BCR/ABL−:

BCR/ABL negative cells

BCR/ABL+:

BCR/ABL positive cells

CA:

Continuous administration

CAT:

Catalase

CML:

Chronic myeloid leukemia

CP:

Chronic phase

ΔΨm:

Mitochondrial transmembrane potential

DMSO:

Dimethyl sulfoxide

EDTA:

Ethylenediaminetetraacetic acid

GSH:

Glutathione

HDC:

Highly damaged cells

IFN:

Interferon

LMA:

Low-melting agarose

MDC:

Middle damaged cells

PBS:

Phosphate-buffer solution

PI-3k:

Phosphatidylinositol-3 kinase

PMN:

Polymorphonuclear cells

Rh123:

Rhodamine 123

RNAse A:

Ribonuclease A

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

STI571:

2-Phenyaminopyrimidine derivative

UC:

Undamaged cells

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Acknowledgments

This paper was supported by grants from CONICET, ANPCyT, and Academia Nacional de Medicina.

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Authors and Affiliations

  1. Depto de Genética, Instituto de Investigaciones Hematológicas “Mariano R. Castex”, Academia de Medicina, J.A. Pacheco de Melo 3081, 1425, Buenos Aires, Argentina

    Luciano Vellón, Marcela González-Cid, Marcelo de Campos Nebel & Irene Larripa

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  1. Luciano Vellón
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  2. Marcela González-Cid
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  3. Marcelo de Campos Nebel
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  4. Irene Larripa
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Correspondence to Irene Larripa.

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Vellón, L., González-Cid, M., Nebel, M.d.C. et al. Additive apoptotic effect of STI571 with the cytoprotective agent amifostine in K-562 cell line. Cancer Chemother Pharmacol 55, 602–608 (2005). https://doi.org/10.1007/s00280-004-0952-5

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  • DOI: https://doi.org/10.1007/s00280-004-0952-5

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