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Aspirin induces apoptosis in human leukemia cells independently of NF-κB and MAPKs through alteration of the Mcl-1/Noxa balance

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Abstract

Aspirin and other non-steroidal anti-inflammatory drugs induce apoptosis in most cell types. In this study we examined the mechanism of aspirin-induced apoptosis in human leukemia cells. We analyzed the role of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs) pathways. Furthermore, we studied the changes induced by aspirin in some genes involved in the control of apoptosis at mRNA level, by performing reverse transcriptase multiplex ligation-dependent probe amplification (RT-MLPA), and at protein level by Western blot. Our results show that aspirin induced apoptosis in leukemia Jurkat T cells independently of NF-κB. Although aspirin induced p38 MAPK and c-Jun N-terminal kinase activation, selective inhibitors of these kinases did not inhibit aspirin-induced apoptosis. We studied the regulation of Bcl-2 family members in aspirin-induced apoptosis. Aspirin increased the mRNA levels of some pro-apoptotic members, such as BIM, NOXA, BMF or PUMA, but their protein levels did not change. In contrast, aspirin decreased the protein levels of Mcl-1. Interestingly, in the presence of aspirin the protein levels of Noxa remained high. This alteration of the Mcl-1/Noxa balance was also found in other leukemia cell lines and primary chronic lymphocytic leukemia cells (CLL). Furthermore, in CLL cells aspirin induced an increase in the protein levels of Noxa. Knockdown of Noxa or Puma significantly attenuated aspirin-induced apoptosis. These results indicate that aspirin induces apoptosis through alteration of the Mcl-1/Noxa balance.

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Abbreviations

NSAIDs:

Non-steroidal anti-inflammatory drugs

NF-κB:

Nuclear factor-κB

MAPK:

Mitogen-activated protein kinase

COX:

Cyclooxygenase

IκB:

Inhibitors of κB

IKKβ:

IκB kinase β

ERK:

Extracellular-regulated kinase

JNK:

c-Jun N-terminal kinase

TPA:

12-O-tetradecanoylphorbol 13-acetate

MAPKAPK 2:

MAPK-activated protein kinase 2

PI:

Propidium iodide

PPARδ:

Peroxisome proliferator-activated receptor δ

EMSA:

Electrophoretic mobility shift assay

ASA:

Aspirin

CHX:

Cycloheximide

RT-MLPA:

Reverse transcriptase multiplex ligation-dependent probe amplification

CLL:

Chronic lymphocytic leukemia

siRNA:

Small interfering RNA

MEFs:

Mouse embryonic fibroblasts

PBLs:

Peripheral blood lymphocytes

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Acknowledgments

We are grateful to Dr. Pura Muñoz and Dr. Francesc Ventura for kindly providing plasmids. We thank Alba Pérez-Perarnau and Camila Rubio for helpful discussions and suggestions. We also thank the Unitat de Biologia and the Unitat de Genòmica of the Serveis Cientificotècnics at the Universitat de Barcelona for their technical support. This study was supported by grants from the Ministerio de Educación y Ciencia and FEDER (SAF2007-60964) and from the Ministerio de Sanidad y Consumo (ISCIII-RETIC RD06/0020) to J.G. D.I.-S. is recipient of fellowship from the José Carreras International Leukemia Foundation (FIJC-07/ESP-FCAJAMADRID). M.d.F. is a recipient of a fellowship from the AGAUR-Generalitat de Catalunya, A.M.C. and D.M.G.-G. are recipients of research fellowships from the Ministerio de Educación y Ciencia. We thank the Language Services at the Universitat de Barcelona for correcting the English of the manuscript.

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The authors declare no competing financial interest.

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Authors and Affiliations

  1. Unitat de Bioquímica, Departament de Ciències Fisiològiques II, IDIBELL-Universitat de Barcelona, Campus de Bellvitge, C/Feixa Llarga s/n, Pavelló de Govern, 4a planta, 08907, L’Hospitalet de Llobregat, Barcelona, Spain

    Daniel Iglesias-Serret, Maria Piqué, Montserrat Barragán, Ana M. Cosialls, Antonio F. Santidrián, Diana M. González-Gironès, Llorenç Coll-Mulet, Mercè de Frias, Gabriel Pons & Joan Gil

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  1. Daniel Iglesias-Serret
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Correspondence to Joan Gil.

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Iglesias-Serret, D., Piqué, M., Barragán, M. et al. Aspirin induces apoptosis in human leukemia cells independently of NF-κB and MAPKs through alteration of the Mcl-1/Noxa balance. Apoptosis 15, 219–229 (2010). https://doi.org/10.1007/s10495-009-0424-9

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