Expression of A2A and A2B Adenosine Receptors in Human Breast Tumors

Document Type : Original Article (s)

Authors

School of Pharmacy and Bioinformatics Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.

Abstract

Background: The expression profile, signal transduction, molecular function and cell growth modulation of adenosine receptor subtypes in the human breast cancer cell lines are described; we decided to investigate the possible roles of adenosine receptors in the human breast tissues. In this study, we used Real-Time (RT) PCR to assess A2A and A2B gene expression in normal and tumoral breast tissues.Methods: Breast tumors and non-neoplastic mammary tissues (n = 15) were collected immediately after mastectomy and stored at -80°C until use. All tumors were histologically confirmed to be breast cancer. Total RNA was extracted and reverse transcribed to cDNA. PCR primers were synthesized from human adenosine receptor cDNA sequences. PCR was performed under optimized condition for each receptor subtype. Amplification of beta-actin mRNA served as control for RT-PCR. The PCR products were separated on 1.5% agarose gels.Finding: To elucidate the expression of A2A and A2B mRNA in breast carcinoma and normal tissues, we compared the level of A2A and A2B mRNA expression by RT-PCR analysis. All breast tumor and normal tissue specimens expressed A2A and A2B adenosine receptor transcripts. We observed that expression of A2B in tumor tissues is 1.36 fold of normal tissues but there was no difference between the expressions of A2A in tumor and normal tissues of the breast, when normalized against that of beta-actin.Conclusion: These results indicated for the first time, to our knowledge, the expression profile of A2A and A2B adenosine receptors in the human breast carcinoma. The present study showed that a high A2B gene expression level was found in breast tumor tissues in comparison with the normal tissues. However, further studies based on the Real-time quantitative RT-PCR are needed to confirm gene expression levels.

Keywords

References

  1. 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(4): 527-52.
  2. Fishman P, Bar-Yehuda S. Pharmacology and therapeutic applications of A3 receptor subtype. Curr Top Med Chem 2003; 3(4): 463-9.
  3. Dhalla AK, Shryock JC, Shreeniwas R, Belardinelli L. Pharmacology and therapeutic applications of A1 adenosine receptor ligands. Curr Top Med Chem 2003; 3(4): 369-85.
  4. Fredholm BB, Cunha RA, Svenningsson P. Pharmacology of adenosine A2A receptors and therapeutic applications. Curr Top Med Chem 2003; 3(4): 413-26.
  5. Volpini R, Costanzi S, Vittori S, Cristalli G, Klotz KN. Medicinal chemistry and pharmacology of A2B adenosine receptors. Curr Top Med Chem 2003; 3(4): 427-43.
  6. Blay J, White TD, Hoskin DW. The extracellular fluid of solid carcinomas contains immunosuppressive concentrations of adenosine. Cancer Res 1997; 57(13): 2602-5.
  7. Spychala J. Tumor-promoting functions of adenosine. Pharmacol Ther 2000; 87(2-3): 161-73.
  8. 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(1): 19-23.
  9. Panjehpour M, Karami-Tehrani F. An adenosine analog (IB-MECA) inhibits anchorage-dependent cell growth of various human breast cancer cell lines. Int J Biochem Cell Biol 2004; 36(8): 1502-9.
  10. Merighi S, Mirandola P, Varani K, Gessi S, Leung E, Baraldi PG et al. A glance at adenosine receptors: novel target for antitumor therapy. Pharmacol Ther 2003; 100(1): 31-48.
  11. Sitkovsky MV. Use of the A(2A) adenosine receptor as a physiological immunosuppressor and to engineer inflammation in vivo. Biochem Pharmacol 2003; 65(4): 493-501.
  12. Dubey RK, Gillespie DG, Jackson EK. A(2B) adenosine receptors stimulate growth of porcine and rat arterial endothelial cells. Hypertension 2002; 39(2 Pt 2): 530-5.
  13. 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(4): 923-33.
  14. National Breast Cancer Centre. Early detection of breast cancer. Position statement. Available at www.nbcc.org. au/resources /documents/ EDP_ earlydetectionposition0804. pdf [Accessed September 2007].
  15. Harirchi I, Ebrahimi M, Zamani N, Jarvandi S, Montazeri A. Breast cancer in Iran: a review of 903 case records. Public Health 2000; 114(2): 143-5.
  16. Mousavi SM, Montazeri A, Mohagheghi MA, Jarrahi AM, Harirchi I, Najafi M et al. Breast cancer in Iran: an epidemiological review. Breast J 2007; 13(4): 383-91.
  17. Sambrook J, Russel DW. Molecular cloning: a laboratory manual. 3rd ed. New York: Cold Spring Harbor; 2001.
  18. Serazin-Leroy V, Denis-Henriot D, Morot M, de Mazancourt P, Giudicelli Y. Semi-quantitative RT-PCR for comparison of mRNAs in cells with different amounts of housekeeping gene transcripts. Mol Cell Probes 1998; 12(5): 283-91.
  19. Dixon AK, Gubitz AK, Sirinathsinghji DJ, Richardson PJ, Freeman TC. Tissue distribution of adenosine receptor mRNAs in the rat. Br J Pharmacol 1996; 118(6): 1461-8.
  20. Panjehpour M, Castro M, Klotz KN. Human breast cancer cell line MDA-MB-231 expresses endogenous A2B adenosine receptors mediating a Ca2+ signal. Br J Pharmacol 2005; 145(2): 211-8.
  21. Panjehpour M, Karami-Tehrani F. Adenosine modulates cell growth in the human breast cancer cells via adenosine receptors. Oncol Res 2007; 16(12): 575-85.
  22. Zeng D, Maa T, Wang U, Feoktistov I, Biggioni I, Belardinelli L. Expression and function of A2B adenosine receptors in the U87MG tumor cells. Drug Dev Res 2003; 58(4): 405-11.
  23. Madi L, Ochaion A, Rath-Wolfson L, Bar-Yehuda S, Erlanger A, Ohana G, et al. The A3 adenosine receptor is highly expressed in tumor versus normal cells: potential target for tumor growth inhibition. Clin Cancer Res 2004; 10(13): 4472-9.
Journal of Isfahan Medical School
Volume 28, Issue 115 - Serial Number 115
September and October 2011
Pages 915-923

Files

History

  • Receive Date: 13 January 2011
  • Revise Date: 29 April 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics