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Caffeic acid phenylethyl ester and MG132, two novel nonconventional chemotherapeutic agents, induce apoptosis of human leukemic cells by disrupting mitochondrial function

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Abstract

The ability to modulate balance between cell survival and death is recognized for its great therapeutic potential. Therefore, research continues to focus on elucidation of cell machinery and signaling pathways that control cell proliferation and apoptosis. Conventional chemotherapeutic agents often have a cytostatic effect over tumor cells. New natural or synthetic chemotherapeutic agents have a wider spectrum of interesting antitumor activities that merit in-depth studies. In the present work, we aimed at characterizing the molecular mechanism leading to induction of cell death upon treatment of the lymphoblastoid cell line PL104 with caffeic acid phenylethyl ester (CAPE), MG132 and two conventional chemotherapeutic agents, doxorubicine (DOX) and vincristine (VCR). Our results showed several apoptotic hallmarks such as phosphatidylserine (PS) exposure on the outer leaflet of the cell membrane, nuclear fragmentation, and increase sub-G1 DNA content after all treatments. In addition, all four drugs downregulated survivin expression. CAPE and both chemotherapeutic agents reduced Bcl-2, while only CAPE and MG132 significantly increased Bax level. CAPE and VCR treatment induced the collapse of mitochondrial membrane potential (∆ψm). All compounds induced cytochrome c release from mitochondrial compartment to cytosol. However, only MG132 caused the translocation of Smac/DIABLO. Except for VCR treatment, all other drugs increased reactive oxygen species (ROS) production level. All treatments induced activation of caspases 3/7, but only CAPE and MG132 led to the activation of caspase 9. In conclusion, our results indicate that CAPE and MG132 treatment of PL104 cells induced apoptosis through the mitochondrial intrinsic pathway, whereas the apoptotic mechanism induced by DOX and VCR may proceed through the extrinsic pathway.

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Abbreviations

CAPE:

Caffeic acid phenylethyl ester

DOX:

Doxorubicine

VCR:

Vincristine

DMSO:

Dimethylsulfoxide

FCS:

Fetal calf serum

PI:

Propidium iodide

DAPI:

4′-6-Diamidino-2-phenylindole

∆ψm:

Mitochondrial membrane potential

ROS:

Reactive oxygen species

H2O2 :

Hydrogen peroxide

AO:

Acridine orange

EB:

Ethidium bromide

IAP:

Inhibitory apoptotic proteins

MMP:

Mitochondrial membrane permeabilization

XIAP:

X-linked inhibitor of apoptosis

cIAP-1:

Cellular inhibitor of apoptosis protein 1

cIAP-2:

Cellular inhibitor of apoptosis protein 2

AML:

Acute myeloid leukemia

ALL:

Acute lymphoblastic leukemia

CML:

Chronic myeloid leukemia

CLL:

Chronic lymphoblastic leukemia

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Acknowledgments

The authors thank Dr. Daniela Ureta (Servicio de Citometría de flujo, Departamento de Microbiología, Inmunología y Biotecnología, Fac. de Fcia. y Bioq., UBA, Argentina) for technical assistance and UBA and CONICET for financial support.

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Authors and Affiliations

  1. Laboratorio de Inmunología Tumoral (LIT), IDEHU-CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires (UBA), Buenos Aires, Argentina

    Victoria Cavaliere, Daniela L. Papademetrio, Susana N. Costantino & Elida M. C. Álvarez

  2. Laboratorio de Immunotoxicologia (LaITO), IDEHU-CONICET, Hospital de Clínicas, José de San Martín, Universidad de Buenos Aires (UBA), Buenos Aires, Argentina

    Tomás Lombardo & Guillermo A. Blanco

Authors
  1. Victoria Cavaliere
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  2. Daniela L. Papademetrio
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  4. Susana N. Costantino
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  5. Guillermo A. Blanco
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  6. Elida M. C. Álvarez
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Corresponding author

Correspondence to Victoria Cavaliere.

Additional information

Victoria Cavaliere, Daniela Papademetrio and Tomás Lombardo are fellows from CONICET.

Guillermo Blanco and Elida Álvarez are members of the National Research Career CONICET.

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Cavaliere, V., Papademetrio, D.L., Lombardo, T. et al. Caffeic acid phenylethyl ester and MG132, two novel nonconventional chemotherapeutic agents, induce apoptosis of human leukemic cells by disrupting mitochondrial function. Targ Oncol 9, 25–42 (2014). https://doi.org/10.1007/s11523-013-0256-y

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  • DOI: https://doi.org/10.1007/s11523-013-0256-y

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