Biopolym. Cell. 2024; 40(4):305-313.
http://dx.doi.org/10.7124/bc.000B08
Molecular Biomedicine
Cytotoxic Effect of Endothelial-Monocyte Activating Polypeptide II on Glioma Cells in vitro
1, 2Shuba I. M., 2Lylo V. V., 2Karpova I. S., 1Glavatskyi O. Ya., 1Kornelyuk A. I.
  1. SI “A.P. Romodanov Neurosurgery Institute, NAMS of Ukraine”
    32, Platona Mayborody Str., Kyiv, Ukraine, 04050
  2. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143

Abstract

Aim. To investigate the in vitro cytotoxic effect of EMAP II on glioma cells by determining the viability of cell culture using the MTT test. Methods. The culture of the human glioma cell line U251MG and the primary cell culture obtained from the malignant glioma tissue fragments after surgical intervention were treated with EMAP II at different concentrations. Cell viability was determined by MTT assay. Results. Cytokine EMAP II exhibits dose–dependent cytotoxic properties in the U251GM cell culture. In the studied concentration range (1.024 nM – 10.0 μM), it exhibits in the MTT test a biphasic effect on cell survival with two statistically significant minima in the concentration range of 640.0 pM (71.6 ± 6.4 %) and 10.0 μM (49.2 ± 11.4 %). The obtained results may indicate that EMAP II can interact with its receptors at ultralow concentrations in a different way than at high concentrations. A dose–dependent cytotoxic effect of EMAP II on glioma cells in the primary culture was shown as well but in the glioblastoma cells this dependence was less pronounced than in the gliomas grade 3 of anaplasia ones. It may be connected with both individual features of patients and genetic differences between glioblastoma and diffuse gliomas grade 3 of anaplasia cells. Conclusions. The complex pattern of dose dependent action of EMAP II on glioma cells may be a result of the multifunctionality of this polypeptide and its ability to interact with various target receptors.
Keywords: cytokine EMAP II, glioma cell line U251GM, primary glioma cell culture, cytotoxicity
Full text: (PDF, in English)

References

[1] Zeng T, Cui D, Gao L. Glioma: an overview of current classifications, characteristics, molecular biology and target therapies. Front Biosci (Landmark Ed). 2015; 20(7):1104-15.
[2] Zozulya YuA, Vasil'yeva IG, Glavatskiy AYa, Rozumenko VD, Lisyaniy NI, Gridina NY. Gliomy golovnogo mozga, Ed. by YuA Zozulya. - K.: UIPK "YeksOb", 2007. - 630 p.
[3] Goodenberger ML, Jenkins RB. Genetics of adult glioma. Cancer Genet. 2012; 205(12):613-21.
[4] Arruebo M, Vilaboa N, Sáez-Gutierrez B, Lambea J, Tres A, Valladares M, González-Fernández A. Assessment of the evolution of cancer treatment therapies. Cancers (Basel). 2011; 3(3):3279-330.
[5] Khranovs'ka NM, Skachkova OV, Horbach OI, Zhukova VM, Hlavats'kyi OYa, Zemskova OV, Khmelnytskyi HV, Shuba IM. The first experience of using dendritic cells immunotherapy in complex treatment of patients with glioblastoma in Ukraine. Clin Oncol. 2019; 34(2):80-6.
[6] Moses MA, Brem H, Langer R. Advancing the field of drug delivery: taking aim at cancer. Cancer Cell. 2003; 4(5):337-41.
[7] Russo S, Cinausero M, Gerratana L, Bozza C, Iacono D, Driol P, Deroma L, Sottile R, Fasola G, Puglisi F. Factors affecting patient's perception of anticancer treatments side-effects: an observational study. Expert Opin Drug Saf. 2014; 13(2):139-50.
[8] Rahman M, Alam K, Hafeez A, Ilyas R, Beg S. Protein-based nanomedicines as anticancer drug delivery platforms. In Micro and Nano Technologies, Nanoformulation Strategies for Cancer Treatment, Elsevier. 2021; 153-69.
[9] Serna N, Sánchez-García L, Unzueta U, Díaz R, Vázquez E, Mangues R, Villaverde A. Protein-Based Therapeutic Killing for Cancer Therapies. Trends Biotechnol. 2018; 36(3):318-35.
[10] Shapira A, Livney YD, Broxterman HJ, Assaraf YG. Nanomedicine for targeted cancer therapy: towards the overcoming of drug resistance. Drug Resist Updat. 2011; 14(3):150-63.
[11] Bar-Zeev M, Livney YD, Assaraf YG. Targeted nanomedicine for cancer therapeutics: Towards precision medicine overcoming drug resistance. Drug Resist Updat. 2017; 31:15-30.
[12] Schwarz MA, Kandel J, Brett J, Li J, Hayward J, Schwarz RE, Chappey O, Wautier JL, Chabot J, Lo Gerfo P, Stern D. Endothelial-monocyte activating polypeptide II, a novel antitumor cytokine that suppresses primary and metastatic tumor growth and induces apoptosis in growing endothelial cells. J Exp Med. 1999; 190(3):341-54.
[13] van Horssen R, Eggermont AM, ten Hagen TL. Endothelial monocyte-activating polypeptide-II and its functions in (patho)physiological processes. Cytokine Growth Factor Rev. 2006; 17(5):339-48.
[14] Schwarz RE, Awasthi N, Konduri S, Cafasso D, Schwarz MA. EMAP II-based antiangiogenic-antiendothelial in vivo combination therapy of pancreatic cancer. Ann Surg Oncol. 2010; 17(5):1442-52.
[15] Ivakhno SS, Kornelyuk AI. Cytokine-like activities of some aminoacyl-tRNA synthetases and auxiliary p43 cofactor of aminoacylation reaction and their role in oncogenesis. Exp Oncol. 2004; 26(4):250-5.
[16] Reznikov AG, Chaykovskaya LV, Polyakova LI, Kornelyuk AI. Antitumor effect of endothelial monocyte-activating polypeptide-II on human prostate adenocarcinoma in mouse xenograft model. Exp Oncol. 2007; 29(4):267-71.
[17] Reznikov AG, Chaykovskaya LV, Polyakova LI, Kornelyuk AI, Grygorenko VN. Cooperative antitumor effect of endothelial-monocyte activating polypeptide II and flutamide on human prostate cancer xenografts. Exp Oncol. 2011; 33(4):231-4.
[18] Shuba IM, Lylo VV, Karpova IS, Glavatskyi OY, Kornelyuk OI. The primary culture of malignant glioma cells as a model for the study of anti-tumor activity of substances. Visnik ukrains'kogo tovaristva genetikiv i selekcioneriv. 2020; 17(2):196-203.
[19] Dubrovsky AL, Brown Jn, Kornelyuk AI, Murray JC, Matsuka GKh. Bacterial Expression of Full Length and Truncated Forms of Cytokine EMAP2 and Cytokine Like Domain of Mammalian Tyrosyl tRNA Synthetase. Biopolym Cell. 2000; 16:229-35.
[20] Kolomiiets LA, Vorobyova NV, Lozhko DM, Zayets VM, Kornelyuk AI. Stabilization of AIMP1/p43 and EMAP II recombinant proteins in the complexes with polysaccharide dextran-70. Pharmacol Rep. 2020; 72(1):238-45.
[21] Stockert JC, Horobin RW, Colombo LL, Blázquez-Castro A. Tetrazolium salts and formazan products in Cell Biology: Viability assessment, fluorescence imaging, and labeling perspectives. Acta Histochem. 2018; 120(3):159-67.
[22] Zaman R, Islam RA, Chowdhury EH. Evolving therapeutic proteins to precisely kill cancer cells. J Control Release. 2022; 351:779-804.
[23] Boohaker RJ, Lee MW, Vishnubhotla P, Perez JM, Khaled AR. The use of therapeutic peptides to target and to kill cancer cells. Curr Med Chem. 2012; 19(22):3794-804.
[24] Schwarz MA, Kandel J, Brett J, Li J, Hayward J, Schwarz RE, Chappey O, Wautier JL, Chabot J, Lo Gerfo P, Stern D. Endothelial-monocyte activating polypeptide II, a novel antitumor cytokine that suppresses primary and metastatic tumor growth and induces apoptosis in growing endothelial cells. J Exp Med. 1999; 190(3):341-54.
[25] Berger AC, Tang G, Alexander HR, Libutti SK. Endothelial monocyte-activating polypeptide II, a tumor-derived cytokine that plays an important role in inflammation, apoptosis, and angiogenesis. J Immunother. 2000; 23(5):519-27.
[26] Berger AC, Alexander HR, Tang G, Wu PS, Hewitt SM, Turner E, Kruger E, Figg WD, Grove A, Kohn E, Stern D, Libutti SK. Endothelial monocyte activating polypeptide II induces endothelial cell apoptosis and may inhibit tumor angiogenesis. Microvasc Res. 2000; 60(1):70-80.
[27] Kayton ML, Libutti SK. Endothelial monocyte activating polypeptide II (EMAP II) enhances the effect of TNF on tumor-associated vasculature. Curr Opin Investig Drugs. 2001; 2(1):136-8.
[28] Lans TE, Van Horssen R, Eggermont AM, Ten Hagen TL. Involvement of endothelial monocyte activating polypeptide II in tumor necrosis factor-alpha-based anti-cancer therapy. Anticancer Res. 2004; 24(4):2243-8.
[29] Crippa L, Gasparri A, Sacchi A, Ferrero E, Curnis F, Corti A. Synergistic damage of tumor vessels with ultra low-dose endothelial-monocyte activating polypeptide-II and neovasculature-targeted tumor necrosis factor-alpha. Cancer Res. 2008; 68(4):1154-61.
[30] Levanets OV, Naidenov VG, Odynets KA, Woodmaska MI, Matsuka GK, Kornelyuk AI. Homology of C-terminal non-catalytic domain of mammalian tyrosyl-tRNAsynthetase with cylokine EMAP II and non-catalytic domains of methionyl- and phenylalanyl-tRNA synthetases. Biopolym Cell. 1997; 13(6): 474-8.
[31] Kornelyuk AI, Tas M, Dubrovsky AL, Murray JC. Cytokine activity of the non-catalytic EMAP-2-like domain of mammalian tyrosyl-tRNA synthetase. Biopolym Cell. 1999; 15(2):168-72.
[32] Chen J, Liu L, Liu Y, Liu X, Qu C, Meng F, Ma J, Lin Y, Xue Y. Low-Dose Endothelial-Monocyte-Activating Polypeptide-II Induced Autophagy by Down-Regulating miR-20a in U-87 and U-251 Glioma Cells. Front Cell Neurosci. 2016; 10:128.
[33] Li Z, Ma J, Liu L, Liu X, Wang P, Liu Y, Li Z, Zheng J, Chen J, Tao W, Xue Y. Endothelial-Monocyte Activating Polypeptide II Suppresses the In Vitro Glioblastoma-Induced Angiogenesis by Inducing Autophagy. Front Mol Neurosci. 2017; 10:208.
[34] Xie H, Xue YX, Liu LB, Liu YH, Wang P. Role of RhoA/ROCK signaling in endothelial-monocyte-activating polypeptide II opening of the blood-tumor barrier: role of RhoA/ROCK signaling in EMAP II opening of the BTB. J Mol Neurosci. 2012; 46(3):666-76.
[35] Li Z, Liu YH, Liu XB, Xue YX, Wang P, Liu LB. Low-dose endothelial monocyte-activating polypeptide-II increases permeability of blood-tumor barrier via a PKC-ζ/PP2A-dependent signaling mechanism. Exp Cell Res. 2015; 331(2):257-66.
[36] Reddy S, Tatiparti K, Sau S, Iyer AK. Recent advances in nano delivery systems for blood-brain barrier (BBB) penetration and targeting of brain tumors. Drug Discov Today. 2021; 26(8):1944-52.
[37] Glavatskyi OY, Zemskova OV, Khmelnytskyi HV, Kardash KA, Shuba IM, Stuley VA. Temozolomide in glioblastoma treatment: 15-year clinical experience and analysis of its efficacy. Exp Oncol. 2020; 42(2):148-56.