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Targeted Oncology

, Volume 9, Issue 1, pp 25–42 | Cite as

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

  • Victoria CavaliereEmail author
  • Daniela L. Papademetrio
  • Tomás Lombardo
  • Susana N. Costantino
  • Guillermo A. Blanco
  • Elida M. C. Álvarez
Original Research

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.

Keywords

Apoptosis Caffeic acid phenylethyl ester (CAPE) MG132 Mitochondria Leukemia 

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

Notes

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.

Conflict of interest

None.

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Copyright information

© Springer-Verlag France 2013

Authors and Affiliations

  • Victoria Cavaliere
    • 1
    Email author
  • Daniela L. Papademetrio
    • 1
  • Tomás Lombardo
    • 2
  • Susana N. Costantino
    • 1
  • Guillermo A. Blanco
    • 2
  • Elida M. C. Álvarez
    • 1
  1. 1.Laboratorio de Inmunología Tumoral (LIT), IDEHU-CONICET, Facultad de Farmacia y BioquímicaUniversidad de Buenos Aires (UBA)Buenos AiresArgentina
  2. 2.Laboratorio de Immunotoxicologia (LaITO), IDEHU-CONICET, Hospital de Clínicas, José de San MartínUniversidad de Buenos Aires (UBA)Buenos AiresArgentina

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