Requirement of Apoptotic Protease-Activating Factor-1 for Bortezomib-Induced Apoptosis but Not for Fas-Mediated Apoptosis in Human Leukemic Cells
Ottosson-Wadlund, Astrid ; Ceder, Rebecca ; Preta, Giulio ; Pokrovskaja, Katja ; Grafstrom, Roland C. ; Heyman, Mats ; Soderhall, Stefan ; Grander, Dan ; Hedenfalk, Ingrid LU
and Robertson, John D.
, et al.
(2013)
In Molecular Pharmacology
83(1).
p.245-255
- Abstract
- Bortezomib is a highly selective inhibitor of the 26S proteasome and has been approved for clinical use in the treatment of relapsing and refractory multiple myeloma and mantle cell lymphoma. Clinical trials are also underway to assess the role of bortezomib in several other human malignancies, including leukemia. However, the mechanism(s) by which bortezomib acts remain to be fully understood. Here, we studied the molecular requirements of bortezomib-induced apoptosis using the human T-cell leukemic Jurkat cells stably transfected with or without shRNA against apoptotic protease-activating factor-1 (Apaf-1). The Apaf-1-deficient Jurkat T cells were resistant to bortezomib-induced apoptosis, as assessed by caspase-3 activity,... (More)
- Bortezomib is a highly selective inhibitor of the 26S proteasome and has been approved for clinical use in the treatment of relapsing and refractory multiple myeloma and mantle cell lymphoma. Clinical trials are also underway to assess the role of bortezomib in several other human malignancies, including leukemia. However, the mechanism(s) by which bortezomib acts remain to be fully understood. Here, we studied the molecular requirements of bortezomib-induced apoptosis using the human T-cell leukemic Jurkat cells stably transfected with or without shRNA against apoptotic protease-activating factor-1 (Apaf-1). The Apaf-1-deficient Jurkat T cells were resistant to bortezomib-induced apoptosis, as assessed by caspase-3 activity, poly(ADP-ribose) polymerase cleavage, phosphatidylserine externalization, and hypodiploid DNA content. In contrast, Apaf-1-deficient cells were sensitive to Fas-induced apoptosis. Bortezomib induced an upregulation of the pro-apoptotic protein Noxa, loss of mitochondrial transmembrane potential, and release of cytochrome c in cells expressing or not expressing Apaf-1. Transient silencing of Apaf-1 expression in RPMI 8402 T-cell leukemic cells also diminished bortezomib-induced apoptosis. Fas-associated death domain (FADD)-deficient Jurkat cells were resistant to Fas-mediated apoptosis yet remained sensitive to bortezomib. Our results show that bortezomib induces apoptosis by regulating pathways that are mechanistically different from those activated upon death receptor ligation. Furthermore, in silico analyses of public transcriptomics data-bases indicated elevated Apaf-1 expression in several hematologic malignancies, including acute lymphoblastic and myeloid leukemia. We also noted variable Apaf-1 expression in a panel of samples from patients with acute lymphoblastic leukemia. Our results suggest that the expression of Apaf-1 may be predictive of the response to proteasome inhibition. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3481454
- author
- Ottosson-Wadlund, Astrid
; Ceder, Rebecca
; Preta, Giulio
; Pokrovskaja, Katja
; Grafstrom, Roland C.
; Heyman, Mats
; Soderhall, Stefan
; Grander, Dan
; Hedenfalk, Ingrid
LU
and Robertson, John D.
, et al. (More)
- Ottosson-Wadlund, Astrid
; Ceder, Rebecca
; Preta, Giulio
; Pokrovskaja, Katja
; Grafstrom, Roland C.
; Heyman, Mats
; Soderhall, Stefan
; Grander, Dan
; Hedenfalk, Ingrid
LU
; Robertson, John D.
and Fadeel, Bengt
(Less)
- organization
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Molecular Pharmacology
- volume
- 83
- issue
- 1
- pages
- 245 - 255
- publisher
- American Society for Pharmacology and Experimental Therapeutics
- external identifiers
-
- wos:000312798900023
- scopus:84871593969
- pmid:23093495
- ISSN
- 1521-0111
- DOI
- 10.1124/mol.112.080788
- language
- English
- LU publication?
- yes
- id
- ab184b07-32a3-42dc-9b0d-93faf28d2247 (old id 3481454)
- date added to LUP
- 2016-04-01 10:10:43
- date last changed
- 2022-01-25 20:31:27
@article{ab184b07-32a3-42dc-9b0d-93faf28d2247,
abstract = {{Bortezomib is a highly selective inhibitor of the 26S proteasome and has been approved for clinical use in the treatment of relapsing and refractory multiple myeloma and mantle cell lymphoma. Clinical trials are also underway to assess the role of bortezomib in several other human malignancies, including leukemia. However, the mechanism(s) by which bortezomib acts remain to be fully understood. Here, we studied the molecular requirements of bortezomib-induced apoptosis using the human T-cell leukemic Jurkat cells stably transfected with or without shRNA against apoptotic protease-activating factor-1 (Apaf-1). The Apaf-1-deficient Jurkat T cells were resistant to bortezomib-induced apoptosis, as assessed by caspase-3 activity, poly(ADP-ribose) polymerase cleavage, phosphatidylserine externalization, and hypodiploid DNA content. In contrast, Apaf-1-deficient cells were sensitive to Fas-induced apoptosis. Bortezomib induced an upregulation of the pro-apoptotic protein Noxa, loss of mitochondrial transmembrane potential, and release of cytochrome c in cells expressing or not expressing Apaf-1. Transient silencing of Apaf-1 expression in RPMI 8402 T-cell leukemic cells also diminished bortezomib-induced apoptosis. Fas-associated death domain (FADD)-deficient Jurkat cells were resistant to Fas-mediated apoptosis yet remained sensitive to bortezomib. Our results show that bortezomib induces apoptosis by regulating pathways that are mechanistically different from those activated upon death receptor ligation. Furthermore, in silico analyses of public transcriptomics data-bases indicated elevated Apaf-1 expression in several hematologic malignancies, including acute lymphoblastic and myeloid leukemia. We also noted variable Apaf-1 expression in a panel of samples from patients with acute lymphoblastic leukemia. Our results suggest that the expression of Apaf-1 may be predictive of the response to proteasome inhibition.}},
author = {{Ottosson-Wadlund, Astrid and Ceder, Rebecca and Preta, Giulio and Pokrovskaja, Katja and Grafstrom, Roland C. and Heyman, Mats and Soderhall, Stefan and Grander, Dan and Hedenfalk, Ingrid and Robertson, John D. and Fadeel, Bengt}},
issn = {{1521-0111}},
language = {{eng}},
number = {{1}},
pages = {{245--255}},
publisher = {{American Society for Pharmacology and Experimental Therapeutics}},
series = {{Molecular Pharmacology}},
title = {{Requirement of Apoptotic Protease-Activating Factor-1 for Bortezomib-Induced Apoptosis but Not for Fas-Mediated Apoptosis in Human Leukemic Cells}},
url = {{http://dx.doi.org/10.1124/mol.112.080788}},
doi = {{10.1124/mol.112.080788}},
volume = {{83}},
year = {{2013}},
}