Selenium Enhances the Apoptotic Efficacy of Docetaxel Through Activation of TRPM2 Channel in DBTRG Glioblastoma Cells
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.
KeywordsApoptosis Docetaxel Glioblastoma Selenium TRPM2 channel
N-(p-amylcinnamoyl) anthranilic acid
Denver Brain Tumor Research Group O5
- DHR 123
Nicotinamide adenine dinucleotide.
Nicotinamide adenine dinucleotide reduced form
Poly (ADP-ribose) polymerase 1
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.
The study was supported by BSN Health, Analysis and Innovation Ltd. Inc. Teknokent, Isparta, Turkey (Project No: 2018-02).
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.
- 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
- 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. https://doi.org/10.1016/j.jtemb.2017.04.012
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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