Hypersensitivity of mtDNA-depleted cells to staurosporine-induced apoptosis: Roles of Bcl-2 downregulation and cathepsin B

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Abstract

We show that mitochondrial DNA (mtDNA)-depleted 143B cells are hypersensitive to staurosporine- induced cell death as evidenced by a more pronounced DNA fragmentation, a stronger activation of caspase-3, an enhanced poly- (ADP-ribose) polymerase-1 (PARP-1) cleavage, and a more dramatic cytosolic release of cytochrome c. We also show that B-cell CLL/ lymphoma-2 (Bcl-2), B-cell lymphoma extra large (Bcl-X L), and myeloid cell leukemia-1 (Mcl-1) are constitutively less abundant in mtDNA-depleted cells, that the inhibition of Bcl-2 and Bcl-X L can sensitize the parental cell line to staurosporine-induced apoptosis, and that overexpression of Bcl-2 or Bcl-X L can prevent the activation of caspase-3 in ρ°143B cells treated with staurosporine. Moreover, the inactivation of cathepsin B with CA074-Me significantly reduced cytochrome c release, caspase-3 activation, PARP-1 cleavage, and DNA fragmentation in mtDNA-depleted cells, whereas the pancaspase inhibitor failed to completely prevent PARP-1 cleavage and DNA fragmentation in these cells, suggesting that caspaseindependent mechanisms are responsible for cell death even if caspases are activated. Finally, we show that cathepsin B is released in the cytosol of ρ° cells in response to staurosporine, suggesting that the absence of mitochondrial activity leads to a facilitated permeabilization of lysosomal membranes in response to staurosporine. © 2011 the American Physiological Society.

Original languageEnglish
Pages (from-to)1090-1106
Number of pages17
JournalAmerican Journal of Physiology - Cell Physiology
Volume300
Issue number5
DOIs
Publication statusPublished - May 2011

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Cathepsin B
Staurosporine
B-Cell Lymphoma
Mitochondrial DNA
Hypersensitivity
Down-Regulation
Apoptosis
DNA Fragmentation
Caspase 3
Cytochromes c
Cell Death
Myeloid Leukemia
Myeloid Cells
Caspases
Cytosol
Cell Line
Membranes
Poly (ADP-Ribose) Polymerase-1

Keywords

  • Lysosome
  • Mitochondrial DNA depletion

Cite this

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title = "Hypersensitivity of mtDNA-depleted cells to staurosporine-induced apoptosis: Roles of Bcl-2 downregulation and cathepsin B",
abstract = "We show that mitochondrial DNA (mtDNA)-depleted 143B cells are hypersensitive to staurosporine- induced cell death as evidenced by a more pronounced DNA fragmentation, a stronger activation of caspase-3, an enhanced poly- (ADP-ribose) polymerase-1 (PARP-1) cleavage, and a more dramatic cytosolic release of cytochrome c. We also show that B-cell CLL/ lymphoma-2 (Bcl-2), B-cell lymphoma extra large (Bcl-X L), and myeloid cell leukemia-1 (Mcl-1) are constitutively less abundant in mtDNA-depleted cells, that the inhibition of Bcl-2 and Bcl-X L can sensitize the parental cell line to staurosporine-induced apoptosis, and that overexpression of Bcl-2 or Bcl-X L can prevent the activation of caspase-3 in ρ°143B cells treated with staurosporine. Moreover, the inactivation of cathepsin B with CA074-Me significantly reduced cytochrome c release, caspase-3 activation, PARP-1 cleavage, and DNA fragmentation in mtDNA-depleted cells, whereas the pancaspase inhibitor failed to completely prevent PARP-1 cleavage and DNA fragmentation in these cells, suggesting that caspaseindependent mechanisms are responsible for cell death even if caspases are activated. Finally, we show that cathepsin B is released in the cytosol of ρ° cells in response to staurosporine, suggesting that the absence of mitochondrial activity leads to a facilitated permeabilization of lysosomal membranes in response to staurosporine. {\circledC} 2011 the American Physiological Society.",
keywords = "Lysosome, Mitochondrial DNA depletion",
author = "Guillaume Rommelaere and S{\'e}bastien Michel and Ludovic Mercy and Antoine Fattaccioli and Catherine Demazy and Noelle Ninane and Andr{\'e}e Houbion and Patricia Renard and Thierry Arnould",
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T1 - Hypersensitivity of mtDNA-depleted cells to staurosporine-induced apoptosis: Roles of Bcl-2 downregulation and cathepsin B

AU - Rommelaere, Guillaume

AU - Michel, Sébastien

AU - Mercy, Ludovic

AU - Fattaccioli, Antoine

AU - Demazy, Catherine

AU - Ninane, Noelle

AU - Houbion, Andrée

AU - Renard, Patricia

AU - Arnould, Thierry

PY - 2011/5

Y1 - 2011/5

N2 - We show that mitochondrial DNA (mtDNA)-depleted 143B cells are hypersensitive to staurosporine- induced cell death as evidenced by a more pronounced DNA fragmentation, a stronger activation of caspase-3, an enhanced poly- (ADP-ribose) polymerase-1 (PARP-1) cleavage, and a more dramatic cytosolic release of cytochrome c. We also show that B-cell CLL/ lymphoma-2 (Bcl-2), B-cell lymphoma extra large (Bcl-X L), and myeloid cell leukemia-1 (Mcl-1) are constitutively less abundant in mtDNA-depleted cells, that the inhibition of Bcl-2 and Bcl-X L can sensitize the parental cell line to staurosporine-induced apoptosis, and that overexpression of Bcl-2 or Bcl-X L can prevent the activation of caspase-3 in ρ°143B cells treated with staurosporine. Moreover, the inactivation of cathepsin B with CA074-Me significantly reduced cytochrome c release, caspase-3 activation, PARP-1 cleavage, and DNA fragmentation in mtDNA-depleted cells, whereas the pancaspase inhibitor failed to completely prevent PARP-1 cleavage and DNA fragmentation in these cells, suggesting that caspaseindependent mechanisms are responsible for cell death even if caspases are activated. Finally, we show that cathepsin B is released in the cytosol of ρ° cells in response to staurosporine, suggesting that the absence of mitochondrial activity leads to a facilitated permeabilization of lysosomal membranes in response to staurosporine. © 2011 the American Physiological Society.

AB - We show that mitochondrial DNA (mtDNA)-depleted 143B cells are hypersensitive to staurosporine- induced cell death as evidenced by a more pronounced DNA fragmentation, a stronger activation of caspase-3, an enhanced poly- (ADP-ribose) polymerase-1 (PARP-1) cleavage, and a more dramatic cytosolic release of cytochrome c. We also show that B-cell CLL/ lymphoma-2 (Bcl-2), B-cell lymphoma extra large (Bcl-X L), and myeloid cell leukemia-1 (Mcl-1) are constitutively less abundant in mtDNA-depleted cells, that the inhibition of Bcl-2 and Bcl-X L can sensitize the parental cell line to staurosporine-induced apoptosis, and that overexpression of Bcl-2 or Bcl-X L can prevent the activation of caspase-3 in ρ°143B cells treated with staurosporine. Moreover, the inactivation of cathepsin B with CA074-Me significantly reduced cytochrome c release, caspase-3 activation, PARP-1 cleavage, and DNA fragmentation in mtDNA-depleted cells, whereas the pancaspase inhibitor failed to completely prevent PARP-1 cleavage and DNA fragmentation in these cells, suggesting that caspaseindependent mechanisms are responsible for cell death even if caspases are activated. Finally, we show that cathepsin B is released in the cytosol of ρ° cells in response to staurosporine, suggesting that the absence of mitochondrial activity leads to a facilitated permeabilization of lysosomal membranes in response to staurosporine. © 2011 the American Physiological Society.

KW - Lysosome

KW - Mitochondrial DNA depletion

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U2 - 10.1152/ajpcell.00037.2010

DO - 10.1152/ajpcell.00037.2010

M3 - Article

C2 - 21068357

VL - 300

SP - 1090

EP - 1106

JO - American Journal of Physiology - Cell Physiology

JF - American Journal of Physiology - Cell Physiology

SN - 0363-6143

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