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The Journal of Membrane Biology

, Volume 249, Issue 1–2, pp 129–140 | Cite as

Synergic Effects of Doxorubicin and Melatonin on Apoptosis and Mitochondrial Oxidative Stress in MCF-7 Breast Cancer Cells: Involvement of TRPV1 Channels

  • Pınar Aslan Koşar
  • Mustafa NazıroğluEmail author
  • İshak Suat Övey
  • Bilal Çiğ
Article

Abstract

Transient receptor transient receptor potential vanilloid 1 (TRPV1) is a Ca2+-permeable channel gated by oxidative stress and capsaicin (CAP) and modulated by melatonin (MEL) and capsazepine (CPZ). A combination of doxorubicin (DOX) and MEL may offer a potential therapy for breast cancer by exerting antitumor and anti-apoptotic effects and modulating Ca2+ influx and TRPV1 activity. We aimed to investigate the effects of MEL and DOX on the oxidative toxicity of MCF-7 human breast cancer cells, in addition to the activity of the TRPV1 channel and apoptosis. The MCF-7 cells were divided into the following six treatment groups: control, incubated with MEL (0.3 mM), incubated with 0.5 μM DOX, incubated with 1 μM DOX, incubated with MEL + 0.5 μM DOX, or incubated with MEL + 1 μM DOX. The intracellular free Ca2+ concentration was higher in the DOX groups than in the control, and the concentration was decreased by MEL. The intracellular free Ca2+ concentration was further increased by treatment with the TRPV1 channel activator CAP (0.01 mM), and it was decreased by the CPZ (0.1 mM). The intracellular production of reactive oxygen species, mitochondrial membrane depolarization, apoptosis level, procaspase 9 and PARP activities, and caspase 3 and caspase 9 activities were higher in the DOX and MEL groups than in the control. Apoptosis and the activity of caspase 9 were further increased in the DOX plus MEL groups. Taken together, the findings indicate that MEL supported the effects of DOX by activation of TRPV1 and apoptosis, as well as by inducing MCF-7 cell death. As the apoptosis and caspase activity of cancer cells increase because of their elevated metabolism, MEL may be useful in supporting their apoptotic capacity.

Keywords

Apoptosis Breast cancer cells Calcium signaling Doxorubicin Mitochondria Oxidative stress 

Abbreviations

[Ca2+]c

Intracellular free Ca2+ concentration

CAP

Capsaicin

CPZ

Capsazepine

DMSO

Dimethyl sulfoxide

DOX

Doxorubicin

EGTA

Ethylene glycol-bis[2-aminoethyl-ether]-N,N,N,N-tetraacetic acid

JC-1,1,1′,3,3′

Tetraethylbenzimidazolylcarbocyanine iodide

PARP

Poly (ADP-ribose) polymerase

ROS

Reactive oxygen species

TRP

Transient receptor potential

TRPM2

Transient receptor potential melastatin 2

TRPV1

Transient receptor potential vanilloid 1

Notes

Acknowledgments

MN formulated the present hypothesis and was responsible for writing the report. BÇ, ISÖ, and PAK were responsible for the analysis of the data.

Financial Support

This study received no financial support.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflicts of interest.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Pınar Aslan Koşar
    • 1
  • Mustafa Nazıroğlu
    • 2
    • 3
    • 4
    Email author
  • İshak Suat Övey
    • 3
    • 4
  • Bilal Çiğ
    • 3
    • 4
  1. 1.Department of Medical Biology, Faculty of MedicineSuleyman Demirel UniversityIspartaTurkey
  2. 2.Department of Biophysics, Faculty of MedicineSuleyman Demirel UniversityIspartaTurkey
  3. 3.Neuroscience Research CenterSuleyman Demirel UniversityIspartaTurkey
  4. 4.Department of Neuroscience, Health Science InstituteSuleyman Demirel UniversityIspartaTurkey

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