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Constitutive presence of cytochrome c in the cytosol of a chemoresistant leukemic cell line

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Abstract

The release of holocytochrome c (cyt c) from mitochondria into the cytosol is reportedly a landmark of the execution phase of apoptosis. As shown here, the P-glycoprotein- (P-gp) expressing K562/ADR cell line (but not the parental K562 cell line) exhibits both cytosolic and mitochondrial cyt c in the absence of any signs of apoptosis. K562/ADR cells were found to be relatively resistant to a variety of different inducers of apoptosis, and blocking the P-gp did not reverse this resistance. The release of cyt c in non-apoptotic K562/ADR cells was not accompanied by that of any other mitochondrial apoptogenic protein, such as AIF or Smac/DIABLO, and was inhibited by Bcl-2 over expression. In addition, using a cell-free system, we show that mitochondria isolated from K562/ADR cells spontaneously released cyt c. These data suggest that cyt c release may be compatible with the preservation of mitochondrial integrity and function, as well as cell proliferation.

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Abbreviations

Ac-DEVD-AMC:

acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin

Ac-LEHD-AMC:

acetyl-Leu-Glu-His-Asp-7-amido-4-methylcoumarin

BSA:

bovine serum albumin

ANT:

Adenine Nucleotide Transporter

apo-cyt c:

apocytochrome c

cyt c:

holocytochrome c

DTT:

dithiothreitol

MDR:

multidrug resistance

PBS:

phosphate buffered saline

P-gp:

P-glycoprotein

PTP:

mitochondrial permeability transition pore

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

VDAC:

Voltage Dependent Anionic Channel

XIAP:

X-chromosome-linked inhibitor of apoptosis protein

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Authors and Affiliations

  1. Equipe 4, UMR 601 INSERM/Université de Nantes, 9 Quai Moncousu

    L. Oliver, M.-T. LeCabellec, K. Meflah & F. M. Vallette

  2. Faculté de Médecine de Nantes, Centre de Microscopie Electronique de l’Ensemble Santé, 44035, Nantes Cedex 01, France

    G. Pradal

  3. CNRS UMR1599, Institut Gustave Roussy, 39 rue Camille-Desmoulins, F-94805, Villejuif, France

    G. Kroemer

  4. Equipe 4, UMR 601 INSERM/Université de Nantes, 9 Quai Moncousu, 44035, Nantes Cedex 01, France

    F. M. Vallette

Authors
  1. L. Oliver
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  2. M.-T. LeCabellec
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  3. G. Pradal
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  4. K. Meflah
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  5. G. Kroemer
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  6. F. M. Vallette
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Correspondence to F. M. Vallette.

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Oliver, L., LeCabellec, MT., Pradal, G. et al. Constitutive presence of cytochrome c in the cytosol of a chemoresistant leukemic cell line. Apoptosis 10, 277–287 (2005). https://doi.org/10.1007/s10495-005-0802-x

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  • DOI: https://doi.org/10.1007/s10495-005-0802-x

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