Neurotoxicity Research

, Volume 35, Issue 4, pp 797–808 | Cite as

Selenium Enhances the Apoptotic Efficacy of Docetaxel Through Activation of TRPM2 Channel in DBTRG Glioblastoma Cells

  • Kemal Ertilav
  • Mustafa Nazıroğlu
  • Zeki Serdar Ataizi
  • Nady BraidyEmail author
Original Article


The rate of mitosis of cancer cells is significantly higher than normal primary cells with increased metabolic needs, which in turn enhances the generation of reactive oxygen species (ROS) production. Higher ROS production is known to increase cancer cell dependence on ROS scavenging systems to counteract the increased ROS. Therapeutic options which selectively modulate the levels of intracellular ROS in cancers are likely candidates for drug discovery. Docetaxel (DTX) has demonstrated antitumor activity in preclinical and clinical studies. It is thought that DTX induces cell death through excessive ROS production and increased Ca2+ entry. The Ca2+ permeable TRPM2 channel is activated by ROS. Selenium (Se) has been previously used to stimulate apoptosis for the treatment of glioblastoma cells resistant to DTX. However, the potential mechanism(s) of the additive effect of DTX on TRPM2 channels in cancer cells remains unclear. The aim of this study was to evaluate the effect of combination therapy of DTX and Se on activation of TRPM2 in DBTRG glioblastoma cells. DBTRG cells were divided into four treatment groups: control, DTX (10 nM for 10 h), Se (1 μM for 10 h), and DTX+Se. Our study showed that apoptosis (Annexin V and propidium iodide), mitochondrial membrane depolarization (JC1), and ROS production levels were increased in DBTRG cells following treatment with Se and DTX respectively. Cell number and viability, and the levels of apoptosis, JC1, ROS, and [Ca2+]i, induced by DTX, were further increased following addition of Se. We also observed an additive increase in the activation of the NAD-dependent DNA repair enzyme poly (ADP-ribose) polymerase-1 (PARP-1) activity, which was accompanied by a decline in its essential substrate NAD+. As well, the Se- and DTX-induced increases in intracellular Ca2+ florescence intensity were decreased following treatment with the TRPM2 antagonist N-(p-amylcinnamoyl) anthranilic acid (ACA). Therefore, combination therapy with Se and DTX may represent an effective strategy for the treatment of glioblastoma cells and may be associated with TRPM2-mediated increases in oxidative stress and [Ca2+]i.


Apoptosis Docetaxel Glioblastoma Selenium TRPM2 channel 



N-(p-amylcinnamoyl) anthranilic acid




Calcium ion


Cyclic ADPR


Cumene hydroperoxide


Denver Brain Tumor Research Group O5

DHR 123

Dihydrorhodamine 123




5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzimidazolylcarbocyanine iodide




Nicotinamide adenine dinucleotide.


Nicotinamide adenine dinucleotide reduced form




Poly (ADP-ribose) polymerase 1


Propidium iodide


Reactive oxygen species




Transient receptor potential ankyrin 1


Transient receptor potential melastatin 2


Transient receptor potential vanilloid 1



The authors wish to thank technician Hulusi Gül (BSN Health, Analysis and Innovation Ltd. Inc. Teknokent, Isparta, Turkey) for their help in performing cell number and cell viability analyses.

Financial Support

The study was supported by BSN Health, Analysis and Innovation Ltd. Inc. Teknokent, Isparta, Turkey (Project No: 2018-02).

Authors’ Contributions

KE, MN, and NB formulated the present hypothesis and MN and NB were responsible for writing the report. MN performed the confocal analyses and the LC-MS analysis. ZSA was responsible for analysis of the data. KE, ZSA, and NB made critical revision of the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Alptekin M, Eroglu S, Tutar E, Sencan S, Geyik MA, Ulasli M, Demiryurek AT, Camci C (2015) Gene expressions of TRP channels in glioblastoma multiforme and relation with survival. Tumour Biol 36(12):9209–9213CrossRefGoogle Scholar
  2. Bao L, Chen SJ, Conrad K, Keefer K, Abraham T, Lee JP, Wang JF, Zhang XQ, Hirschler-Laszkiewicz I, Wang HG, Dovat S, Gans B, Madesh M, Cheung JY, Miller BA (2016) Depletion of the human ion channel TRPM2 in neuroblastoma demonstrates its key role in cell survival through modulation of mitochondrial reactive oxygen species and bioenergetics. J Biol Chem 291(47):24449–24464CrossRefGoogle Scholar
  3. Berthier S, Arnaud J, Champelovier P, Col E, Garrel C, Cottet C, Boutonnat J, Laporte F, Faure P, Hazane-Puch F (2017) Anticancer properties of sodium selenite in human glioblastoma cell cluster spheroids. J Trace Elem Med Biol 44:161–176.
  4. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem 53:452–458Google Scholar
  5. Braidy N, Berg J, Clement J, Poljak A, Grant R et al. (2018) Role of NAD+ and related precursors as therapeutic targets for age-related degenerative diseases: rationale, biochemistry, pharmacokinetics, and outcomes. Antioxidant and Redox SignallingGoogle Scholar
  6. Bustamante S, Jayasena T, Richani D, Gilchrist R, Wu L et al (2018) Quantifying the cellular NAD+ metabolome using a tandem liquid chromatography mass spectrometry approach. Metabolomics 14:15CrossRefGoogle Scholar
  7. Demirdas A, Naziroglu M, Ovey IS (2017) Duloxetine reduces oxidative stress, apoptosis, and Ca(2+) entry through modulation of TRPM2 and TRPV1 channels in the hippocampus and dorsal root ganglion of rats. Mol Neurobiol 54(6):4683–4695CrossRefGoogle Scholar
  8. Elf AK, Bernhardt P, Hofving T, Arvidsson Y, Forssell-Aronsson E, Wängberg B, Nilsson O, Johanson V (2017) NAMPT inhibitor GMX1778 enhances the efficacy of 177Lu-DOTATATE treatment of neuroendocrine tumors. J Nucl Med 58(2):288–292CrossRefGoogle Scholar
  9. Esposito E, Impellizzeri D, Mazzon E, Fakhfouri G, Rahimian R et al (2012) The NAMPT inhibitor FK866 reverts the damage in spinal cord injury. J Neuroinflammation 9:66Google Scholar
  10. Gallego-Yerga L, Posadas I, de la Torre C, Ruiz-Almansa J, Sansone F, Ortiz Mellet C, Casnati A, García Fernández JM, Ceña V (2017) Docetaxel-loaded nanoparticles assembled from beta-cyclodextrin/calixarene giant surfactants: physicochemical properties and cytotoxic effect in prostate cancer and glioblastoma cells. Front Pharmacol 8:249CrossRefGoogle Scholar
  11. Gao H, Zhang S, Yang Z, Cao S, Jiang X, Pang Z (2014) In vitro and in vivo intracellular distribution and anti-glioblastoma effects of docetaxel-loaded nanoparticles functioned with IL-13 peptide. Int J Pharm 466(1–2):8–17CrossRefGoogle Scholar
  12. Ghoochani A, Hatipoglu Majernik G, Sehm T, Wach S, Buchfelder M, Taubert H, Eyupoglu IY, Savaskan N (2016) Cabazitaxel operates anti-metastatic and cytotoxic via apoptosis induction and stalls brain tumor angiogenesis. Oncotarget 7(25):38306–38318CrossRefGoogle Scholar
  13. Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2(1):48–58CrossRefGoogle Scholar
  14. Hazane-Puch F, Arnaud J, Trocme C, Faure P, Laporte F et al (2016) Sodium selenite decreased HDAC activity, cell proliferation and induced apoptosis in three human glioblastoma cells. Anti Cancer Agents Med Chem 16(4):490–500CrossRefGoogle Scholar
  15. Hua H, Zhang X, Mu H, Meng Q, Jiang Y, Wang Y, Lu X, Wang A, Liu S, Zhang Y, Wan Z, Sun K (2018) RVG29-modified docetaxel-loaded nanoparticles for brain-targeted glioma therapy. Int J Pharm 543(1–2):179–189CrossRefGoogle Scholar
  16. Huang K, Bian D, Jiang B, Zhai Q, Gao N, Wang R (2017) TRPA1 contributed to the neuropathic pain induced by docetaxel treatment. Cell Biochem Funct 35(3):141–143CrossRefGoogle Scholar
  17. Joshi DC, Bakowska JC (2011) Determination of mitochondrial membrane potential and reactive oxygen species in live rat cortical neurons. J Vis Exp(51)Google Scholar
  18. Ju RJ, Mu LM, Li XT, Li CQ, Cheng ZJ et al. (2018) Development of functional docetaxel nanomicelles for treatment of brain glioma. Artif Cells Nanomed Biotechnol 46:sup1, 1180–1190Google Scholar
  19. Lev-Ram V, Ellisman MH (1995) Axonal activation-induced calcium transients in myelinating Schwann cells, sources, and mechanisms. J Neurosci 15(4):2628–2637CrossRefGoogle Scholar
  20. Naziroglu M (2009) Role of selenium on calcium signaling and oxidative stress-induced molecular pathways in epilepsy. Neurochem Res 34(12):2181–2191CrossRefGoogle Scholar
  21. Naziroglu M (2017) Activation of TRPM2 and TRPV1 channels in dorsal root ganglion by NADPH oxidase and protein kinase C molecular pathways: a patch clamp study. J Mol Neurosci 61(3):425–435CrossRefGoogle Scholar
  22. Naziroglu M, Braidy N (2017) Thermo-sensitive TRP channels: novel targets for treating chemotherapy-induced peripheral pain. Front Physiol 8:1040CrossRefGoogle Scholar
  23. Naziroglu M, Luckhoff A (2008a) A calcium influx pathway regulated separately by oxidative stress and ADP-ribose in TRPM2 channels: single channel events. Neurochem Res 33(7):1256–1262CrossRefGoogle Scholar
  24. Naziroglu M, Luckhoff A (2008b) Effects of antioxidants on calcium influx through TRPM2 channels in transfected cells activated by hydrogen peroxide. J Neurol Sci 270(1–2):152–158CrossRefGoogle Scholar
  25. Naziroglu M, Karaoglu 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(2–3):221–230CrossRefGoogle Scholar
  26. Naziroglu M, Tokat S, Demirci S (2012a) Role of melatonin on electromagnetic radiation-induced oxidative stress and Ca2+ signaling molecular pathways in breast cancer. J Recept Signal Transduct Res 32(6):290–297CrossRefGoogle Scholar
  27. Naziroglu M, Yildiz K, Tamturk B, Erturan I, Flores-Arce M (2012b) Selenium and psoriasis. Biol Trace Elem Res 150(1–3):3–9CrossRefGoogle Scholar
  28. Nur G, Naziroglu M, Deveci HA (2017) Synergic prooxidant, apoptotic and TRPV1 channel activator effects of alpha-lipoic acid and cisplatin in MCF-7 breast cancer cells. J Recept Signal Transduct Res 37(6):569–577CrossRefGoogle Scholar
  29. Park SO, Yoo YB, Kim YH, Baek KJ, Yang JH, Choi PC, Lee JH, Lee KR, Park KS (2015) Effects of combination therapy of docetaxel with selenium on the human breast cancer cell lines MDA-MB-231 and MCF-7. Ann Surg Treat Res 88(2):55–62CrossRefGoogle Scholar
  30. Saikali S, Avril T, Collet B, Hamlat A, Bansard JY, Drenou B, Guegan Y, Quillien V (2007) Expression of nine tumour antigens in a series of human glioblastoma multiforme: interest of EGFRvIII, IL-13Ralpha2, gp100 and TRP-2 for immunotherapy. J Neuro-Oncol 81(2):139–148CrossRefGoogle Scholar
  31. Sakalli Cetin E, Naziroglu M, Cig B, Ovey IS, Aslan Kosar P (2017) Selenium potentiates the anticancer effect of cisplatin against oxidative stress and calcium ion signaling-induced intracellular toxicity in MCF-7 breast cancer cells: involvement of the TRPV1 channel. J Recept Signal Transduct Res 37(1):84–93CrossRefGoogle Scholar
  32. Shi K, Zhou J, Zhang Q, Gao H, Liu Y, Zong T, He Q (2015) Arginine-glycine-aspartic acid-modified lipid-polymer hybrid nanoparticles for docetaxel delivery in glioblastoma multiforme. J Biomed Nanotechnol 11(3):382–391CrossRefGoogle Scholar
  33. Siegel RL, Miller KD, Jemal A (2018) Cancer statistics, 2018. CA Cancer J Clin 68(1):7–30CrossRefGoogle Scholar
  34. Tabaczar S, Koceva-Chyla A, Matczak K, Gwozdzinski K (2010) Molecular mechanisms of antitumor activity of taxanes. I. Interaction of docetaxel with microtubules. Postepy Hig Med Dosw (Online) 64:568–581Google Scholar
  35. Tanimura K, Uchino J, Tamiya N, Kaneko Y, Yamada T, Yoshimura K, Takayama K (2018) Treatment rationale and design of the RAMNITA study: a phase II study of the efficacy of docetaxel + ramucirumab for non-small cell lung cancer with brain metastasis. Medicine (Baltimore) 97(23):e11084CrossRefGoogle Scholar
  36. Uguz AC, Naziroglu M, Espino J, Bejarano I, Gonzalez D et al (2009) Selenium modulates oxidative stress-induced cell apoptosis in human myeloid HL-60 cells through regulation of calcium release and caspase-3 and -9 activities. J Membr Biol 232(1–3):15–23CrossRefGoogle Scholar
  37. Uslusoy F, Naziroglu M, Cig B (2017) Inhibition of the TRPM2 and TRPV1 channels through Hypericum perforatum in sciatic nerve injury-induced rats demonstrates their key role in apoptosis and mitochondrial oxidative stress of sciatic nerve and dorsal root ganglion. Front Physiol 8:335CrossRefGoogle Scholar
  38. Vercelli C, Barbero R, Cuniberti B, Odore R, Re G (2015) Expression and functionality of TRPV1 receptor in human MCF-7 and canine CF.41 cells. Vet Comp Oncol 13(2):133–142CrossRefGoogle Scholar
  39. Wang Q, Huang L, Yue J (2017) Oxidative stress activates the TRPM2-Ca(2+)-CaMKII-ROS signaling loop to induce cell death in cancer cells. Biochim Biophys Acta 1864(6):957–967CrossRefGoogle Scholar
  40. Wrobel JK, Wolff G, Xiao R, Power RF, Toborek M (2016) Dietary selenium supplementation modulates growth of brain metastatic tumors and changes the expression of adhesion molecules in brain microvessels. Biol Trace Elem Res 172(2):395–407CrossRefGoogle Scholar
  41. Xu HL, Mao KL, Lu CT, Fan ZL, Yang JJ, Xu J, Chen PP, ZhuGe DL, Shen BX, Jin BH, Xiao J, Zhao YZ (2016) An injectable acellular matrix scaffold with absorbable permeable nanoparticles improves the therapeutic effects of docetaxel on glioblastoma. Biomaterials 107:44–60CrossRefGoogle Scholar
  42. Yakubov E, Buchfelder M, Eyupoglu IY, Savaskan NE (2014) Selenium action in neuro-oncology. Biol Trace Elem Res 161(3):246–254CrossRefGoogle Scholar
  43. Zhang XQ, Lu JT, Jiang WX, Lu YB, Wu M, Wei EQ, Zhang WP, Tang C (2015) NAMPT inhibitor and metabolite protect mouse brain from cryoinjury through distinct mechanisms. Neuroscience 291:230–240CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Kemal Ertilav
    • 1
  • Mustafa Nazıroğlu
    • 2
    • 3
  • Zeki Serdar Ataizi
    • 4
  • Nady Braidy
    • 5
    Email author
  1. 1.Department of Neurosurgery, Faculty of MedicineSuleyman Demirel UniversityIspartaTurkey
  2. 2.Neuroscience Research CenterSuleyman Demirel UniversityIspartaTurkey
  3. 3.Drug Discovery and Development Research Group in Neuroscience, BSN Health, Analysis and Innovation, Goller Bolgesi TeknokentiSuleyman Demirel UniversityIspartaTurkey
  4. 4.Department of NeurosurgeryYunus Emre State HospitalEskişehirTurkey
  5. 5.Centre for Healthy Brain Ageing, School of PsychiatryUniversity of New South WalesSydneyAustralia

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