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Inhibitory effect of temozolomide on apoptosis induction of cinnamaldehyde in human glioblastoma multiforme T98G cell line

  • Hedieh Abband (Department of Anatomy, Cellular and Molecular Research Center, School of Medicine, Guilan University of Medical Sciences) ;
  • Sara Dabirian (Department of Pharmaceutical Biotechnology, School of Pharmacy, Guilan University of Medical Sciences) ;
  • Adele Jafari (Department of Physiology, School of Medicine, Guilan University of Medical Sciences) ;
  • Mehran Nasiri (Department of Anatomy, Cellular and Molecular Research Center, School of Medicine, Guilan University of Medical Sciences) ;
  • Ebrahim Nasiri (Department of Anatomy, Cellular and Molecular Research Center, School of Medicine, Guilan University of Medical Sciences)
  • Received : 2023.06.05
  • Accepted : 2023.09.19
  • Published : 2024.03.31

Abstract

Glioblastoma is the most common primary malignant brain tumor in adults. Temozolomide (TMZ) is an FDA-approved drug used to treat this type of cancer. Cinnamaldehyde (CIN) is a derivative of cinnamon extract and makes up 99% of it. The aim of this study was to investigate the in vitro combined effect of CIN and TMZ on human glioblastoma multiforme T98G cell line viability. In this study, we used 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-tertazolium bromide (MTT) method to evaluate the extent of IC50, acridine orange, Giemsa and Hoechst staining to evaluate the manner of apoptosis and the Western blotting method to examine the expression change of apoptotic proteins. Our results show that TMZ has an inhibitory effect on CIN when both used in combination at concentrations of 300 and 100 µM (P<0.05) and has a cytotoxic effect when used alone at the same concentrations (P<0.05). The western blotting result showed that TMZ at concentrations of 2,000 and 1,000 µM significantly increased Bax expression and decreased Bcl2 expression (P<0.05), indicating that TMZ induced apoptosis through the mitochondrial pathway. However, CIN had no effect on Bax and Bcl2 expressions, thus causing apoptosis from another pathway. Also, the Bax:Bcl2 expression ratio at concentrations combined was lower than that for TMZ 1,000 µM and higher than that for CIN 150 and 100 µM (P<0.05), which confirms the inhibitory effect of TMZ on CIN. From the present study, we conclude that TMZ in combination with CIN has an inhibitory effect on increasing the cytotoxicity rate.

Keywords

Acknowledgement

We greatly appreciate the support extended by Dr. Adele Jafari at Cellular and Molecular Research Center and Vice-Chancellor for Research of the Guilan University of Medical Sciences (IR.GUMS.REC.1394.233).

References

  1. Ostrom QT, Gittleman H, Truitt G, Boscia A, Kruchko C, Barnholtz-Sloan JS. CBTRUS statistical report: primary brain and other central nervous system tumors diagnosed in the United States in 2011-2015. Neuro Oncol 2018;20(suppl_4):iv1-86. Erratum in: Neuro Oncol 2021;23:508-22.  https://doi.org/10.1093/neuonc/noy171
  2. Hanif F, Muzaffar K, Perveen K, Malhi SM, Simjee ShU. Glioblastoma multiforme: a review of its epidemiology and pathogenesis through clinical presentation and treatment. Asian Pac J Cancer Prev 2017;18:3-9. 
  3. Ostrom QT, Gittleman H, Liao P, Rouse C, Chen Y, Dowling J, Wolinsky Y, Kruchko C, Barnholtz-Sloan J. CBTRUS statistical report: primary brain and central nervous system tumors diagnosed in the United States in 2007-2011. Neuro Oncol 2014;16(Suppl 4):iv1-63.  https://doi.org/10.1093/neuonc/nou223
  4. Preusser M, de Ribaupierre S, Wohrer A, Erridge SC, Hegi M, Weller M, Stupp R. Current concepts and management of glioblastoma. Ann Neurol 2011;70:9-21.  https://doi.org/10.1002/ana.22425
  5. Stupp R, Brada M, van den Bent MJ, Tonn JC, Pentheroudakis G. High-grade glioma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 2014;25 Suppl 3:iii93-101.  https://doi.org/10.1093/annonc/mdu050
  6. Lee SY. Temozolomide resistance in glioblastoma multiforme. Genes Dis 2016;3:198-210.  https://doi.org/10.1016/j.gendis.2016.04.007
  7. Alonso MM, Gomez-Manzano C, Bekele BN, Yung WK, Fueyo J. Adenovirus-based strategies overcome temozolomide resistance by silencing the O6-methylguanine-DNA methyltransferase promoter. Cancer Res 2007;67:11499-504.  https://doi.org/10.1158/0008-5472.CAN-07-5312
  8. Ramirez YP, Mladek AC, Phillips RM, Gynther M, Rautio J, Ross AH, Wheelhouse RT, Sakaria JN. Evaluation of novel imidazotetrazine analogues designed to overcome temozolomide resistance and glioblastoma regrowth. Mol Cancer Ther 2015;14:111-9.  https://doi.org/10.1158/1535-7163.MCT-14-0113
  9. Park I, Mukherjee J, Ito M, Chaumeil MM, Jalbert LE, Gaensler K, Ronen SM, Nelson SJ, Pieper RO. Changes in pyruvate metabolism detected by magnetic resonance imaging are linked to DNA damage and serve as a sensor of temozolomide response in glioblastoma cells. Cancer Res 2014;74:7115-24.  https://doi.org/10.1158/0008-5472.CAN-14-0849
  10. Smalley S, Chalmers AJ, Morley SJ. mTOR inhibition and levels of the DNA repair protein MGMT in T98G glioblastoma cells. Mol Cancer 2014;13:144. 
  11. Cen L, Carlson BL, Pokorny JL, Mladek AC, Grogan PT, Schroeder MA, Decker PA, Anderson SK, Giannini C, Wu W, Ballman KV, Kitange GJ, Sarkaria JN. Efficacy of protracted temozolomide dosing is limited in MGMT unmethylated GBM xenograft models. Neuro Oncol 2013;15:735-46.  https://doi.org/10.1093/neuonc/not010
  12. Melguizo C, Prados J, Gonzalez B, Ortiz R, Concha A, Alvarez PJ, Madeddu R, Perazzoli G, Oliver JA, Lopez R, Rodriguez-Serrano F, Aranega A. MGMT promoter methylation status and MGMT and CD133 immunohistochemical expression as prognostic markers in glioblastoma patients treated with temozolomide plus radiotherapy. J Transl Med 2012;10:250. 
  13. Sangal A. Role of cinnamon as beneficial antidiabetic food adjunct: a review. Adv Appl Sci Res 2011;2:440-50. 
  14. Vangalapati M, Sree Satya N, Surya Prakash DV, Avanigadda S. A review on pharmacological activities and clinical effects of cinnamon species. Res J Pharm Biol Chem Sci 2012;3:653-63. 
  15. Koh WS, Yoon SY, Kwon BM, Jeong TC, Nam KS, Han MY. Cinnamaldehyde inhibits lymphocyte proliferation and modulates T-cell differentiation. Int J Immunopharmacol 1998;20:643-60.  https://doi.org/10.1016/S0192-0561(98)00064-2
  16. Kwon BM, Lee SH, Choi SU, Park SH, Lee CO, Cho YK, Sung ND, Bok SH. Synthesis and in vitro cytotoxicity of cinnamaldehydes to human solid tumor cells. Arch Pharm Res 1998;21:147-52.  https://doi.org/10.1007/BF02974019
  17. Shaughnessy DT, Setzer RW, DeMarini DM. The antimutagenic effect of vanillin and cinnamaldehyde on spontaneous mutation in Salmonella TA104 is due to a reduction in mutations at GC but not AT sites. Mutat Res 2001;480-481:55-69.  https://doi.org/10.1016/S0027-5107(01)00169-5
  18. Ka H, Park HJ, Jung HJ, Choi JW, Cho KS, Ha J, Lee KT. Cinnamaldehyde induces apoptosis by ROS-mediated mitochondrial permeability transition in human promyelocytic leukemia HL-60 cells. Cancer Lett 2003;196:143-52.  https://doi.org/10.1016/S0304-3835(03)00238-6
  19. Ng LT, Wu SJ. Antiproliferative activity of cinnamomum cassia constituents and effects of pifithrin-alpha on their apoptotic signaling pathways in Hep G2 cells. Evid Based Complement Alternat Med 2011;2011:492148. 
  20. Kanzawa T, Kondo Y, Ito H, Kondo S, Germano I. Induction of autophagic cell death in malignant glioma cells by arsenic trioxide. Cancer Res 2003;63:2103-8. 
  21. Kanzawa T, Germano IM, Komata T, Ito H, Kondo Y, Kondo S. Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ 2004;11:448-57.  https://doi.org/10.1038/sj.cdd.4401359
  22. Jakubowicz-Gil J, Langner E, Badziul D, Wertel I, Rzeski W. Apoptosis induction in human glioblastoma multiforme T98G cells upon temozolomide and quercetin treatment. Tumour Biol 2013;34:2367-78.  https://doi.org/10.1007/s13277-013-0785-0
  23. Pasi F, Persico MG, Buroni FE, Aprile C, Hodolic M, Corbella F, Nano R, Facoetti A, Lodola L. Uptake of 18F-FET and 18F-FCH in human glioblastoma T98G cell line after irradiation with photons or carbon ions. Contrast Media Mol Imaging 2017;2017:6491674. 
  24. Niknezhad F, Sayad-Fathi S, Karimzadeh A, Ghorbani-Anarkooli M, Yousefbeyk F, Nasiri E. Improvement in histology, enzymatic activity, and redox state of the liver following administration of Cinnamomum zeylanicum bark oil in rats with established hepatotoxicity. Anat Cell Biol 2019;52:302-11.  https://doi.org/10.5115/acb.18.180
  25. Figul M, Soling A, Dong HJ, Chou TC, Rainov NG. Combined effects of temozolomide and the ribonucleotide reductase inhibitors didox and trimidox in malignant brain tumor cells. Cancer Chemother Pharmacol 2003;52:41-6.  https://doi.org/10.1007/s00280-003-0611-2
  26. Chen JC, Hsieh PS, Chen SM, Hwang JH. Effects of cinnamaldehyde on the viability and expression of chemokine receptor genes in temozolomide-treated glioma cells. In Vivo 2020;34:595-9.  https://doi.org/10.21873/invivo.11812
  27. Wu SJ, Ng LT, Lin CC. Cinnamaldehyde-induced apoptosis in human PLC/PRF/5 cells through activation of the proapoptotic Bcl-2 family proteins and MAPK pathway. Life Sci 2005;77:938-51.  https://doi.org/10.1016/j.lfs.2005.02.005
  28. Wu SJ, Ng LT. MAPK inhibitors and pifithrin-alpha block cinnamaldehyde-induced apoptosis in human PLC/PRF/5 cells. Food Chem Toxicol 2007;45:2446-53.  https://doi.org/10.1016/j.fct.2007.05.032
  29. Wu SJ, Ng LT, Lin CC. Effects of vitamin E on the cinnamaldehyde-induced apoptotic mechanism in human PLC/PRF/5 cells. Clin Exp Pharmacol Physiol 2004;31:770-6.  https://doi.org/10.1111/j.1440-1681.2004.04091.x
  30. Zhang JH, Liu LQ, He YL, Kong WJ, Huang SA. Cytotoxic effect of trans-cinnamaldehyde on human leukemia K562 cells. Acta Pharmacol Sin 2010;31:861-6.  https://doi.org/10.1038/aps.2010.76
  31. Grootjans S, Hassannia B, Delrue I, Goossens V, Wiernicki B, Dondelinger Y, Bertrand MJ, Krysko DV, Vuylsteke M, Vandenabeele P, Vanden Berghe T. A real-time fluorometric method for the simultaneous detection of cell death type and rate. Nat Protoc 2016;11:1444-54.  https://doi.org/10.1038/nprot.2016.085
  32. Hitomi J, Christofferson DE, Ng A, Yao J, Degterev A, Xavier RJ, Yuan J. Identification of a molecular signaling network that regulates a cellular necrotic cell death pathway. Cell 2008;135:1311-23.  https://doi.org/10.1016/j.cell.2008.10.044
  33. Richards R, Schwartz HR, Honeywell ME, Stewart MS, Cruz-Gordillo P, Joyce AJ, Landry BD, Lee MJ. Drug antagonism and single-agent dominance result from differences in death kinetics. Nat Chem Biol 2020;16:791-800.  https://doi.org/10.1038/s41589-020-0510-4
  34. Chio CC, Tai YT, Mohanraj M, Liu SH, Yang ST, Chen RM. Honokiol enhances temozolomide-induced apoptotic insults to malignant glioma cells via an intrinsic mitochondrion-dependent pathway. Phytomedicine 2018;49:41-51.  https://doi.org/10.1016/j.phymed.2018.06.012
  35. Sur P, Sribnick EA, Patel SJ, Ray SK, Banik NL. Dexamethasone decreases temozolomide-induced apoptosis in human gliobastoma T98G cells. Glia 2005;50:160-7. https://doi.org/10.1002/glia.20168