Advertisement

Tumor Biology

, Volume 34, Issue 2, pp 1085–1095 | Cite as

Adenosine induces cell cycle arrest and apoptosis via cyclinD1/Cdk4 and Bcl-2/Bax pathways in human ovarian cancer cell line OVCAR-3

  • Saeid Shirali
  • Mahmoud Aghaei
  • Mahdi Shabani
  • Mojtaba Fathi
  • Majid Sohrabi
  • Marzieh Moeinifard
Research Article

Abstract

Adenosine is a regulatory molecule with widespread physiological effects in almost every cells and acts as a potent regulator of cell growth. Adenosine has been shown to inhibit cell growth and induce apoptosis in the several cancer cells via caspase activation and Bcl-2/Bax pathway. The present study was designed to understand the mechanism underlying adenosine-induced apoptosis in the OVCAR-3 human ovarian cancer cells. MTT viability, BrdU and cell counting assays were used to study the cell proliferation effect of adenosine in presence of adenosine deaminase inhibitor and the nucleoside transporter inhibitor. Cell cycle analysis, propidium iodide and annexin V staining, caspase-3 activity assay, cyclinD1, Cdk4, Bcl-2 and Bax protein expressions were assessed to detect apoptosis. Adenosine significantly inhibited cell proliferation in a concentration-dependent manner in OVCAR-3 cell line. Adenosine induced cell cycle arrest in G0/G1 phase via Cdk4/cyclinD1-mediated pathway. Adenosine induced apoptosis, which was determined by Annexin V-FITC staining and increased sub-G1 population. Moreover, down-regulation of Bcl-2 protein expression, up-regulation of Bax protein expression and activation of caspase-3 were observed in response to adenosine treatment. The results of this study suggest that extracellular adenosine induced G1 cell cycle arrest and apoptosis in ovarian cancer cells via cyclinD1/ Cdk4 and Bcl-2/Bax pathways and caspase-3 activation. These data might suggest that adenosine could be used as an agent for the treatment of ovarian cancer.

Keywords

Ovarian cancer Adenosine Cell cycle arrest Apoptosis 

Notes

Acknowledgment

This work was supported by a grant from Chalous Branch, Islamic Azad University, Chalous, Iran.

Conflicts of interest

None

References

  1. 1.
    Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, et al. Cancer statistics. CA Cancer J Clin. 2008;58:71–96.PubMedCrossRefGoogle Scholar
  2. 2.
    Chien JR, Aletti G, Bell DA, Keeney GL, Shridhar V, Hartmann LC. Molecular pathogenesis and therapeutic targets in epithelial ovarian cancer. J Cell Biochem. 2007;102:117–29.CrossRefGoogle Scholar
  3. 3.
    Aletti GD, Gallenberg MM, Cliby WA, Jatoi A, Hartmann LC. Current management strategies for ovarian cancer. Mayo Clin Proc. 2007;82:751–70.PubMedGoogle Scholar
  4. 4.
    Gessi S, Merighi S, Sacchetto V, Simioni C, Borea PA. Adenosine receptors and cancer. Biochim Biophys Acta. 2011;1808:1400–12.PubMedCrossRefGoogle Scholar
  5. 5.
    Schneider C, Wiendl H, Ogilive A. Biophasic cytotoxic mechanism of extracellulat ATP on U-937 human histiocytic leukemia cells: involvement of adenosine generation. Biochim Biophys Acta. 2001;1538:190–205.PubMedCrossRefGoogle Scholar
  6. 6.
    Di Iorio P, Kleywegt S, Ciccarelli R, Traversa U, Andrew CM, Crocker CE, et al. Mechanisms of apoptosis induced by purine nucleosides in astrocytes. Glia. 2002;38:179–90.PubMedCrossRefGoogle Scholar
  7. 7.
    Panjehpour M, Karami-Tehrani F. Adenosine modulates cell growth in the human breast cancer cells via adenosine receptors. Oncol Res. 2007;16:575–85.PubMedCrossRefGoogle Scholar
  8. 8.
    Gessi S, Merighi S, Varani K, Cattabriga E, Benini A, Mirandola P, et al. Adenosine receptors in colon carcinoma tissues and colon tumoral cell lines: focus on the A (3) adenosine subtype. J Cell Physiol. 2007;211:826–36.PubMedCrossRefGoogle Scholar
  9. 9.
    Tanaka Y, Yoshihara K, Tsuyuki M, Kamiya T. Apoptosis induced by adenosine in human leukemia HL-60 cells. Exp Cell Res. 1994;213:242–52.PubMedCrossRefGoogle Scholar
  10. 10.
    Merighi S, Mirandola P, Milani D, Varani K, Gessi S, Klotz KN, et al. Adenosine receptors as mediators of both cell proliferation and cell death of cultured human melanoma cells. J Invest Dermatol. 2002;119:923–33.PubMedCrossRefGoogle Scholar
  11. 11.
    Hashemi M, Karami-Tehrani F, Ghavami S, Maddika S, Los M. Adenosine and deoxyadenosine induces apoptosis in oestrogen receptor-positive and -negative human breast cancer cells via the intrinsic pathway. Cell Prolif. 2005;38:269–85.PubMedCrossRefGoogle Scholar
  12. 12.
    Salami S, Karami-Tehrani F. Biochemical studies of apoptosis induced by tamoxifen in estrogen receptor positive and negative breast cancer cell lines. Clin Biochem. 2003;36(4):247–53.PubMedCrossRefGoogle Scholar
  13. 13.
    Aghaei M, Karami-Tehrani F, Salami S, Atri M. Diagnostic value of adenosine deaminase activity in benign and malignant breast tumors. Arch Med Res. 2009;41:14–8.CrossRefGoogle Scholar
  14. 14.
    Schorl C, Sedivy JM. Analysis of cell cycle phases and progression in cultured mammalian cells. Methods. 2007;41:143–50.PubMedCrossRefGoogle Scholar
  15. 15.
    Ghavami S, Rashedi I, Dattilo BM, Eshraghi M, Chazin WJ, Hashemi M, et al. S100A8/A9 at low concentration promotes tumor cell growth via RAGE ligation and MAP-kinase dependent pathway. J Leukoc Biol. 2008;83:1484–92.PubMedCrossRefGoogle Scholar
  16. 16.
    Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991;139:271–9.PubMedCrossRefGoogle Scholar
  17. 17.
    Ghavami S, Eshragi M, Ande SR, Chazin WJ, Klonisch T, Halayko AJ, et al. S100A8/A9 induces autophagy and apoptosis via ROS-mediated cross-talk between mitochondria and lysosomes that involves BNIP3. Cell Res. 2010;20:314–31.PubMedCrossRefGoogle Scholar
  18. 18.
    Aghaei M, Panjehpour M, Karami-tehrani F, Salami S. Molecular mechanism of A3 adenosine receptor-induced G1 cell cycle arrest and apoptosis in androgen-dependent and independent prostate cancer cell lines: involvement of intrinsic pathway. J Cancer Res Clin Oncol. 2011;137:1511–23.PubMedCrossRefGoogle Scholar
  19. 19.
    Ghavami S, Kerkhoff C, Los M, Hashemi M, Sorg C, Karami-Tehrani F. Mechanism of apoptosis induced by S100A8/A9 in colon cancer cell lines: the role of ROS and the effect of metal ions. J Leuk Biol. 2004;76:169–75.CrossRefGoogle Scholar
  20. 20.
    Fredholm BB, IJzerman AP, Jacobson KA, Klotz KN, Linden J. International Union of Pharmacology. XXV. Nomenclature and classification of adenosine receptors. Pharmacol Rev. 2001;53:527–52.PubMedGoogle Scholar
  21. 21.
    Fishman P, Yehuda SB, Ohana G, Pathak S, Wasserman L, Barer F, et al. Adenosine acts as an inhibitor of lymphoma cell growth: a major role for the A3 adenosine receptor. Euro J Cancer. 2000;36:1452–8.CrossRefGoogle Scholar
  22. 22.
    Aghaei M, Karami-Tehrani F, Panjehpour M, Salami S, Fallahian F. Adenosine induces cell-cycle arrest and apoptosis in androgen-dependent and -independent prostate cancer cell lines, LNcap-FGC-10, DU-145, and PC3. Prostate. 2012;72:361–75.PubMedCrossRefGoogle Scholar
  23. 23.
    Kim SJ, Min HY, Chung HJ, Park EJ, Hong JY, Kang YJ, et al. Inhibition of cell proliferation through cell cycle arrest and apoptosis by thio-Cl-IB-MECA, a novel A3 adenosine receptor agonist, in human lung cancer cells. Cancer Lett. 2008;264:309–15.PubMedCrossRefGoogle Scholar
  24. 24.
    Ohana G, Bar-Yehuda S, Barer F, Fishman P. Differential effect of adenosine on tumor and normal cell growth: focus on the A3 adenosine receptor. J Cell Physiol. 2001;186:19–23.PubMedCrossRefGoogle Scholar
  25. 25.
    Meininger CJ, Schelling ME, Granger HJ. Adenosine and hypoxia stimulate proliferation and migration of endothelial cells. Am J Physiol. 1998;255:554–62.Google Scholar
  26. 26.
    Grant MB, Davis MI, Caballero S, Feoktistov I, Biaggioni I, Belardinelli L. Proliferation, migration, and ERK activation in human retinal endothelial cells through A(2B) adenosine receptor stimulation. Invest Ophthalmol Vis Sci. 2001;42:2068–73.PubMedGoogle Scholar
  27. 27.
    Abbracchio MP, Camurri A, Ceruti S, Cattabeni F, Falzano L, Giammarioli AM, et al. The A3 adenosine receptor induces cytoskeleton rearrangement in human astrocytoma cells via a specific action on Rho proteins. Ann N Y Acad Sci. 2001;939:63–73.PubMedCrossRefGoogle Scholar
  28. 28.
    Kohno Y, Sei Y, Koshiba M, Kim HO, Jacobson KA. Induction of apoptosis in HL-60 human promyelocytic leukemia cells by adenosine A(3) receptor agonists. Biochem Biophys Res Commun. 1996;219:904–10.PubMedCrossRefGoogle Scholar
  29. 29.
    Cory S, Adams JM. The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer. 2002;2:647–56.PubMedCrossRefGoogle Scholar
  30. 30.
    Appel E, Kazimirsky G, Ashkenazi E, Kim SG, Jacobson KA, Brodien C. Roles of BCL-2 and caspase 3 in the adenosine A3 receptor-induced apoptosis. J Mol Neurosci. 2001;17:285–92.PubMedCrossRefGoogle Scholar
  31. 31.
    Wu LF, Li GP, Feng JL, Pu ZJ. Molecular mechanisms of adenosine-induced apoptosis in human HepG2 cells. Acta Pharmacol Sin. 2006;27:477–84.PubMedCrossRefGoogle Scholar
  32. 32.
    Zhang YX, Yu SB, Ou-Yang JP, Xia D, Wang M, Li JR. Effect of protein kinase C alpha, caspase-3, and survivin on apoptosis of oral cancer cells induced by staurosporine. Acta Pharmacol Sin. 2005;26:1365–72.PubMedCrossRefGoogle Scholar
  33. 33.
    Kang CM, Sun Y, Jang IS, Park SC. Thymidine-dependent attenuation of the mitochondrial apoptotic pathway in adenosine induced apoptosis of HL-60 cell. J Cancer Res Clin Oncol. 2001;12:570–6.CrossRefGoogle Scholar
  34. 34.
    Tai CJ, Chang SJ, Chien LY, Leung PC, Tzeng CR. Adenosine triphosphate induces activation of caspase-3 in apoptosis of human granulosa-luteal cells. Endocr J. 2005;52:327–35.PubMedCrossRefGoogle Scholar
  35. 35.
    Saito M, Yaguchi T, Yasuda Y, Nakano T, Nishizaki T. Adenosine suppresses CW2 human colonic cancer growth by inducing apoptosis via A1 adenosine receptors. Cancer Lett. 2010;290:211–5.PubMedCrossRefGoogle Scholar
  36. 36.
    Shieh DE, Chen YY, Yen MH, Chiang LC, Lin CC. Emodininduced apoptosis through p53-dependent pathway in human hepatoma cells. Life Sci. 2004;74:2279–90.PubMedCrossRefGoogle Scholar
  37. 37.
    Dolle RE, Hoyer D, Prasad CV, Schmidt SJ, Helaszek CT, Miller RE, et al. P1 aspartate-based peptide alpha-((2,6-dichlorobenzoyl)oxy)methyl ketones as potent time-dependent inhibitors of interleukin-1 beta-converting enzyme. J Med Chem. 1994;37(5):563–4.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2013

Authors and Affiliations

  • Saeid Shirali
    • 1
  • Mahmoud Aghaei
    • 2
  • Mahdi Shabani
    • 3
  • Mojtaba Fathi
    • 4
  • Majid Sohrabi
    • 5
  • Marzieh Moeinifard
    • 4
  1. 1.Department of Laboratory Sciences, Chalous BranchIslamic Azad UniversityChalousIran
  2. 2.Department of Clinical Biochemistry, School of Pharmacy and Isfahan Pharmaceutical Sciences Research CenterIsfahan University of Medical SciencesIsfahanIran
  3. 3.Department of Antibody and Antigen Engendering, Monoclonal Antibody Research Center, Avicenna Research InstituteAcademic Center for Education, Culture and Research (ACECR)TehranIran
  4. 4.Department of Clinical Biochemistry, School of Medical ScienceTarbiat Modares UniversityTehranIran
  5. 5.Department of Bacteriology, School of Medical ScienceTarbiat Modares UniversityTehranIran

Personalised recommendations