Investigation the Effect of Adenosine A1 Receptor Agonist and Antagonist on P53 Gene Expression, and Apoptosis Pathways and Rate in U87Mg Multiform Glioblastoma

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Assistant Professor, Department of Anatomical Sciences, School of Medicine, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

3 Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 Associate Professor, Molecular Medicine Research Center, Hormozgan Health Institute, Hormozgan University of Medical Science, Bandar Abbas, Iran

5 Assistant Professor, Department of Biology and Anatomical Sciences, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

Abstract

Background: Improper prognosis in brain cancers requires new treatments. Using family of purinergic receptors with confirmed apoptotic effect can be beneficial. As the role of type A1 receptor in multiform glioblastoma in relation to the P53 gene and apoptotic pathways is nor reported, we studied the role of agonist (N6-cyclopentyladenosine or CPA) and antagonist (8-cyclopentyl-1,3-dipropylxanthine or DPCPX) of this receptor on cell apoptosis and also expression of P53 genes and caspases 7, 8, and 9.Methods: In this study, MTT assay was used to investigate the rate of cellular proliferation, and flowcytometry method with annexin and Pi was also used to investigate early and late cell apoptosis. To evaluate the internal and external apoptotic pathways expression of P53 genes and caspases 7, 8, and 9, real-time reverse transcription polymerase chain reaction (real-time RT PCR) was used.Findings: The treatment of U87Mg cells with DPCPX increased the expression of P53 gene. Expression of caspase 7 as an executive caspase and caspase 9 as a caspase of the mitochondrial pathway of apoptosis increased, but no expression change was observed in the caspase 8 gene.Conclusion: The results of MTT and flowcytometry showed that DPCPX, in addition to suppressing cell proliferation, stimulated apoptosis in U87Mg cells. Inhibition of adenosine A1 receptors by stimulating the expression of genes involved in apoptotic pathways, especially mitochondrial pathway genes, suppressed cell proliferation and induced apoptosis in U87Mg cells.

Keywords

References

  1. Young A, Mittal D, Stagg J, Smyth MJ. Targeting cancer-derived adenosine: New therapeutic approaches. Cancer Discov 2014; 4(8): 879-88.
  2. Goldschneider D, Mehlen P. Dependence receptors: A new paradigm in cell signaling and cancer therapy. Oncogene 2010; 29(13): 1865-82.
  3. Gessi S, Varani K, Merighi S, Morelli A, Ferrari D, Leung E, et al. Pharmacological and biochemical characterization of A3 adenosine receptors in Jurkat T cells. Br J Pharmacol 2001; 134(1): 116-26.
  4. Sek K, Molck C, Stewart GD, Kats L, Darcy PK, Beavis PA. Targeting adenosine receptor signaling in cancer immunotherapy. Int J Mol Sci 2018; 19(12).
  5. Merighi S, Battistello E, Giacomelli L, Varani K, Vincenzi F, Borea PA, et al. Targeting A3 and A2A adenosine receptors in the fight against cancer. Expert Opin Ther Targets 2019; 23(8): 669-78.
  6. Jaafaru MS, Nordin N, Rosli R, Shaari K, Bako HY, Noor NM, et al. Prospective role of mitochondrial apoptotic pathway in mediating GMG-ITC to reduce cytotoxicity in H2O2-induced oxidative stress in differentiated SH-SY5Y cells. Biomed Pharmacother 2019; 119: 109445.
  7. Timofeev O, Klimovich B, Schneikert J, Wanzel M, Pavlakis E, Noll J, et al. Residual apoptotic activity of a tumorigenic p53 mutant improves cancer therapy responses. EMBO J 2019; 38(20): e102096.
  8. Yue X, Zhao Y, Xu Y, Zheng M, Feng Z, Hu W. Mutant p53 in Cancer: Accumulation, gain-of-function, and therapy. J Mol Biol 2017; 429(11): 1595-606.
  9. Bykov VJN, Eriksson SE, Bianchi J, Wiman KG. Targeting mutant p53 for efficient cancer therapy. Nat Rev Cancer 2018; 18(2): 89-102.
  10. Benit CP, Kerkhof M, Duran-Pena A, Vecht CJ. Seizures as complications in cancer. In: Schiff D, Arrillaga I, Wen PY, editors. Cancer neurology in clinical practice: Neurological complications of cancer and its treatment. Cham, Switzerland: Springer International Publishing; 2018. p. 153-69.
  11. Pearson JRD, Regad T. Targeting cellular pathways in glioblastoma multiforme. Signal Transduction and Targeted Therapy 2017; 2(1): 17040.
  12. Vlacich G, Tsien CI. High-Grade gliomas. In: Chang EL, Brown PD, Lo SS, Sahgal A, Suh JH, editors. Adult CNS radiation oncology: Principles and practice. Cham, Switzerland: Springer International Publishing; 2018. p. 83-102.
  13. Hanif F, Muzaffar K, Perveen K, Malhi SM, Simjee S. Glioblastoma multiforme: A review of its epidemiology and pathogenesis through clinical presentation and treatment. Asian Pac J Cancer Prev 2017; 18(1): 3-9.
  14. Batash R, Asna N, Schaffer P, Francis N, Schaffer M. Glioblastoma multiforme, diagnosis and treatment; recent literature review. Curr Med Chem 2017; 24(27): 3002-9.
  15. Ceruti S, Abbracchio MP. Adenosine signaling in glioma cells. Adv Exp Med Biol 2020; 1202: 13-33.
  16. Niechi I, Uribe-Ojeda A, Erices JI, Torres A, Uribe D, Rocha JD, et al. Adenosine depletion as a new strategy to decrease glioblastoma stem-like cells aggressiveness. Cells 2019; 8(11).
  17. Jafari SM, Joshaghani HR, Panjehpour M, Aghaei M. A2B adenosine receptor agonist induces cell cycle arrest and apoptosis in breast cancer stem cells via ERK1/2 phosphorylation. Cell Oncol (Dordr) 2018; 41(1): 61-72.
  18. Oakes E, Anderson A, Cohen-Gadol A, Hundley HA. Adenosine deaminase that acts on RNA 3 (ADAR3) binding to glutamate receptor subunit B Pre-mRNA inhibits RNA editing in glioblastoma. J Biol Chem 2017; 292(10): 4326-35.
  19. Leone RD, Emens LA. Targeting adenosine for cancer immunotherapy. J Immunother Cancer 2018; 6(1): 57.
  20. Vigano S, Alatzoglou D, Irving M, Menetrier-Caux C, Caux C, Romero P, et al. Targeting adenosine in cancer immunotherapy to enhance T-cell function. Front Immunol 2019; 10: 925.
  21. Sai K, Yang D, Yamamoto H, Fujikawa H, Yamamoto S, Nagata T, et al. A(1) adenosine receptor signal and AMPK involving caspase-9/-3 activation are responsible for adenosine-induced RCR-1 astrocytoma cell death. Neurotoxicology 2006; 27(4): 458-67.
  22. Yasuda Y, Saito M, Yamamura T, Yaguchi T, Nishizaki T. Extracellular adenosine induces apoptosis in Caco-2 human colonic cancer cells by activating caspase-9/-3 via A(2a) adenosine receptors. J Gastroenterol 2009; 44(1): 56-65.
  23. Chen Y, Yang SH, Hueng DY, Syu JP, Liao CC, Wu YC. Cordycepin induces apoptosis of C6 glioma cells through the adenosine 2A receptor-p53-caspase-7-PARP pathway. Chem Biol Interact 2014; 216: 17-25.
  24. Fianco G, Mongiardi MP, Levi A, De Luca T, Desideri M, Trisciuoglio D, et al. Caspase-8 contributes to angiogenesis and chemotherapy resistance in glioblastoma. Elife 2017; 6.
  25. Saito M, Yaguchi T, Yasuda Y, Nakano T, Nishizaki T. Adenosine suppresses CW2 human colonic cancer growth by inducing apoptosis via A(1) adenosine receptors. Cancer Lett 2010; 290(2): 211-5.
  26. Lin Z, Yin P, Reierstad S, O'Halloran M, Coon VJ, Pearson EK, et al. Adenosine A1 receptor, a target and regulator of estrogen receptoralpha action, mediates the proliferative effects of estradiol in breast cancer. Oncogene 2010; 29(8): 1114-22.
  27. Saitoh M, Nagai K, Nakagawa K, Yamamura T, Yamamoto S, Nishizaki T. Adenosine induces apoptosis in the human gastric cancer cells via an intrinsic pathway relevant to activation of AMP-activated protein kinase. Biochem Pharmacol 2004; 67(10): 2005-11.
Journal of Isfahan Medical School
Volume 38, Issue 601 - Serial Number 601
November and December 2021
Pages 875-881

Files

History

  • Receive Date: 30 April 2020
  • Revise Date:
  • Accept Date: 06 April 2022

Share

How to cite

Statistics