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

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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.

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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

References

  • Bejarano I, Espino J, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2011) Pro-oxidant effect of melatonin in tumour leucocytes: relation with its cytotoxic and pro-apoptotic effects. Basic Clin Pharmacol Toxicol 108:14–20

    Article  CAS  PubMed  Google Scholar 

  • Caterina MJ, Schumacher MA, Tominaga M, Rosen TA, Levine JD, Julius D (1997) The capsaicin receptor: a heat-activated ion channel in the pain pathway. Nature 389(6653):816–824

    Article  CAS  PubMed  Google Scholar 

  • Celik O, Nazıroğlu M (2012) Melatonin modulates apoptosis and TRPM2 channels in transfected cells activated by oxidative stress. Physiol Behav 107:458–465

    Article  CAS  PubMed  Google Scholar 

  • Chen SJ, Hoffman NE, Shanmughapriya S, Bao L, Keefer K, Conrad K, Merali S, Takahashi Y, Abraham T, Hirschler-Laszkiewicz I, Wang J, Zhang XQ, Song J, Barrero C, Shi Y, Kawasawa YI, Bayerl M, Sun T, Barbour M, Wang HG, Madesh M, Cheung JY, Miller BA (2014) A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α. J Biol Chem 289:36284–36302

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Çiğ B, Nazıroğlu M (2015) Investigation of the effects of distance from sources on apoptosis, oxidative stress and cytosolic calcium accumulation via TRPV1 channels induced by mobile phones and Wi-Fi in breast cancer cells. Biochim Biophys Acta 1848:2756–2765

    Article  PubMed  Google Scholar 

  • Dziegiel P, Podhorska-Okolow M, Zabel M (2008) Melatonin: adjuvant therapy of malignant tumors. Med Sci Monit 14:RA64–RA70

    CAS  PubMed  Google Scholar 

  • Ekmekcioglu C (2006) Melatonin receptors in humans: biological role and clinical relevance. Biomed Pharmacother 60:97–108

    Article  CAS  PubMed  Google Scholar 

  • Ekmekcioglu C (2014) Expression and putative functions of melatonin receptors in malignant cells and tissues. Wien Med Wochenschr 164:472–478

    Article  PubMed  Google Scholar 

  • Espino J, Bejarano I, Redondo PC, Rosado JA, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2010) Melatonin reduces apoptosis induced by calcium signaling in human leukocytes: evidence for the involvement of mitochondria and Bax activation. J Membr Biol 233:105–118

    Article  CAS  PubMed  Google Scholar 

  • Espino J, Rodríguez AB, Pariente JA (2013) The inhibition of TNF-α-induced leucocyte apoptosis by melatonin involves membrane receptor MT1/MT2 interaction. J Pineal Res 54:442–452

    CAS  PubMed  Google Scholar 

  • Gilliam LA, Fisher-Wellman KH, Lin CT, Maples JM, Cathey BL, Neufer PD (2013) The anticancer agent doxorubicin disrupts mitochondrial energy metabolism and redox balance in skeletal muscle. Free Radic Biol Med 65:988–996

    Article  CAS  PubMed  Google Scholar 

  • González-Flores D, Rodríguez AB, Pariente JA (2014) TNFα-induced apoptosis in human myeloid cell lines HL-60 and K562 is dependent of intracellular ROS generation. Mol Cell Biochem 390:281–287

    Article  PubMed  Google Scholar 

  • Govender J, Loos B, Marais E, Engelbrecht AM (2014) Mitochondrial catastrophe during doxorubicin-induced cardiotoxicity: a review of the protective role of melatonin. J Pineal Res 57:367–380

    Article  CAS  PubMed  Google Scholar 

  • Grynkiewicz C, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    CAS  PubMed  Google Scholar 

  • Gupta S, Sharma B (2014) Pharmacological benefits of agomelatine and vanillin in experimental model of Huntington’s disease. Pharmacol Biochem Behav 122:122–135

    Article  CAS  PubMed  Google Scholar 

  • Heemskerk JW, Feijge MA, Henneman L, Rosing J, Hemker HC (1997) The Ca2+-mobilizing potency of alpha-thrombin and thrombin receptor-activating peptide on human platelets concentration and time effects of thrombin-induced Ca2+ signalling. Eur J Biochem 249:547–555

    Article  CAS  PubMed  Google Scholar 

  • Inoue S, Setoyama Y, Odaka A (2014) Doxorubicin treatment induces tumor cell death followed by immunomodulation in a murine neuroblastoma model. Exp Ther Med 7:703–708

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ip SW, Lan SH, Lu HF, Huang AC, Yang JS, Lin JP, Huang HY, Lien JC, Ho CC, Chiu CF, Wood W, Chung JG (2012) Capsaicin mediates apoptosis in human nasopharyngeal carcinoma NPC-TW 039 cells through mitochondrial depolarization and endoplasmic reticulum stress. Hum Exp Toxicol 31(6):539–549

    Article  CAS  PubMed  Google Scholar 

  • Koçkar MC, Nazıroğlu M, Celik O, Tola HT, Bayram D, Koyu A (2010) N-acetylcysteine modulates doxorubicin-induced oxidative stress and antioxidant vitamin concentrations in liver of rats. Cell Biochem Funct 28(8):673–677

    Article  PubMed  Google Scholar 

  • Köse SA, Nazıroğlu M (2015) N-acetyl cysteine reduces oxidative toxicity, apoptosis, and calcium entry through TRPV1 channels in the neutrophils of patients with polycystic ovary syndrome. Free Radic Res 49:338–346

    Article  PubMed  Google Scholar 

  • Kumar VS, Gopalakrishnan A, Nazıroğlu M, Rajanikant GK (2014) Calcium ion: the key player in cerebral ischemia. Curr Med Chem 21:2065–2075

    Article  PubMed  Google Scholar 

  • Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91:479–489

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Kim CN, Yang J, Jemmerson R, Wang X (1996) Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 86:147–157

    Article  CAS  PubMed  Google Scholar 

  • Nazıroğlu M (2007) New molecular mechanisms on the activation of TRPM2 channels by oxidative stress and ADP-ribose. Neurochem Res 32:1990–2001

    Article  PubMed  Google Scholar 

  • Nazıroğlu M (2012) Molecular role of catalase on oxidative stress-induced Ca(2+) signaling and TRP cation channel activation in nervous system. J Recept Signal Transduct Res 32:134–141

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Karaoğlu A, Aksoy AO (2004) Selenium and high dose vitamin E administration protects cisplatin-induced oxidative damage to renal, liver and lens tissues in rats. Toxicology 195:221–230

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Çelik Ö, Özgül C, Çiğ B, Doğan S, Bal R, Gümral N, Rodríguez AB, Pariente JA (2012) Melatonin modulates wireless (2.45 GHz)-induced oxidative injury through TRPM2 and voltage gated Ca(2+) channels in brain and dorsal root ganglion in rat. Physiol Behav 105:683–692

    Article  PubMed  Google Scholar 

  • Nemeth C, Humpeler S, Kallay E, Mesteri I, Svoboda M, Rögelsperger O, Klammer N, Thalhammer T, Ekmekcioglu C (2011) Decreased expression of the melatonin receptor 1 in human colorectal adenocarcinomas. J Biol Regul Homeost Agents 25:531–542

    CAS  PubMed  Google Scholar 

  • Özdemir ÜS, Nazıroğlu M, Şenol N, Ghazizadeh V (2015) Hypericum perforatum attenuates spinal cord injury-induced oxidative stress and apoptosis in the dorsal root ganglion of rats: involvement of TRPM2 and TRPV1 channels. Mol Neurobiol 23:1–12

    Google Scholar 

  • Radogna F, Paternoster L, De Nicola M, Cerella C, Ammendola S, Bedini A, Tarzia G, Aquilano K, Ciriolo M, Ghibelli L (2009) Rapid and transient stimulation of intracellular reactive oxygen species by melatonin in normal and tumor leukocytes. Toxicol Appl Pharmacol 239:37–45

    Article  CAS  PubMed  Google Scholar 

  • Reiter RJ, Tan DX, Osuna C, Gitto E (2000) Actions of melatonin in the reduction of oxidative stress: a review. J Biomed Sci 7:444–458

    Article  CAS  PubMed  Google Scholar 

  • Reuss S, Disque-Kaiser U, Binzen U, Greffrath W, Peschke E (2010) ‘TRPing’ synaptic ribbon function in the rat pineal gland: neuroendocrine regulation involves the capsaicin receptor TRPV1. Neuroendocrinology 92:133–142

    Article  CAS  PubMed  Google Scholar 

  • Sag CM, Köhler AC, Anderson ME, Backs J, Maier LS (2011) CaMKII-dependent SR Ca leak contributes to doxorubicin-induced impaired Ca handling in isolated cardiac myocytes. J Mol Cell Cardiol 51:749–759

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sag CM, Wagner S, Maier LS (2013) Role of oxidants on calcium and sodium movement in healthy and diseased cardiac myocytes. Free Radic Biol Med 63:338–349

    Article  CAS  PubMed  Google Scholar 

  • Shi Y (2002) Mechanisms of caspase activation and inhibition during apoptosis. Mol Cell 2002(9):459–470

    Article  Google Scholar 

  • Song N, Kim AJ, Kim HJ, Jee HJ, Kim M, Yoo YH, Yun J (2012) Melatonin suppresses doxorubicin-induced premature senescence of A549 lung cancer cells by ameliorating mitochondrial dysfunction. J Pineal Res 53:335–343

    Article  CAS  PubMed  Google Scholar 

  • Sung B, Prasad S, Ravindran J, Yadav VR, Aggarwal BB (2012) Capsazepine, a TRPV1 antagonist, sensitizes colorectal cancer cells to apoptosis by TRAIL through ROS-JNK-CHOP-mediated upregulation of death receptors. Free Radic Biol Med 53(10):1977–1987

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Susankova K, Tousova K, Vyklicky L, Teisinger J, Vlachova V (2006) Reducing and oxidizing agents sensitize heat-activated vanilloid receptor (TRPV1) current. Mol Pharmacol 70:383–394

    CAS  PubMed  Google Scholar 

  • Uğuz AC, Nazıroğlu M, Espino J, Bejarano I, Gonzalez D, Rodriguez AB, Pariente JA (2009) Selenium modulates oxidative stress induced cell apoptosis in human myeloid HL-60 cells via regulation of caspase-3, -9 and calcium influx. J Membr Biol 232:15–23

    Article  PubMed  Google Scholar 

  • Uğuz AC, Cig B, Espino J, Bejarano I, Nazıroğlu M, Rodríguez AB, Pariente JA (2012) Melatonin potentiates chemotherapy-induced cytotoxicity and apoptosis in rat pancreatic tumor cells. J Pineal Res 53:91–98

    Article  PubMed  Google Scholar 

  • Vercelli C, Barbero R, Cuniberti B, Racca S, Abbadessa G, Piccione F, Re G (2014) Transient receptor potential vanilloid 1 expression and functionality in mcf-7 cells: a preliminary investigation. J Breast Cancer 17:332–338

    Article  PubMed  PubMed Central  Google Scholar 

  • Wu TT, Peters AA, Tan PT, Roberts-Thomson SJ, Monteith GR (2014) Consequences of activating the calcium-permeable ion channel TRPV1 in breast cancer cells with regulated TRPV1 expression. Cell Calcium 56(2):59–67

    Article  CAS  PubMed  Google Scholar 

  • Yürüker V, Nazıroğlu M, Şenol N (2015) Reduction in traumatic brain injury-induced oxidative stress, apoptosis, and calcium entry in rat hippocampus by melatonin: possible involvement of TRPM2 channels. Metab Brain Dis 30:223–231

    Article  PubMed  Google Scholar 

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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.

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This study received no financial support.

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Authors and Affiliations

  1. Department of Medical Biology, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey

    Pınar Aslan Koşar

  2. Department of Biophysics, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey

    Mustafa Nazıroğlu

  3. Neuroscience Research Center, Suleyman Demirel University, Isparta, Turkey

    Mustafa Nazıroğlu, İshak Suat Övey & Bilal Çiğ

  4. Department of Neuroscience, Health Science Institute, Suleyman Demirel University, Isparta, Turkey

    Mustafa Nazıroğlu, İshak Suat Övey & Bilal Çiğ

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  1. Pınar Aslan Koşar
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  2. Mustafa Nazıroğlu
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  3. İshak Suat Övey
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  4. Bilal Çiğ
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Correspondence to Mustafa Nazıroğlu.

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Koşar, P.A., Nazıroğlu, M., Övey, İ.S. et al. Synergic Effects of Doxorubicin and Melatonin on Apoptosis and Mitochondrial Oxidative Stress in MCF-7 Breast Cancer Cells: Involvement of TRPV1 Channels. J Membrane Biol 249, 129–140 (2016). https://doi.org/10.1007/s00232-015-9855-0

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  • DOI: https://doi.org/10.1007/s00232-015-9855-0

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