Purinergic Signalling

, Volume 9, Issue 2, pp 271–280 | Cite as

A2B adenosine receptor blockade inhibits growth of prostate cancer cells

  • Qiang Wei
  • Stefano Costanzi
  • Ramachandran Balasubramanian
  • Zhan-Guo Gao
  • Kenneth A. JacobsonEmail author
Original Article


The role of the A2B adenosine receptor (AR) in prostate cell death and growth was studied. The A2B AR gene expression quantified by real-time quantitative RT-PCR and Western blot analysis was the highest among four AR subtypes (A1, A2A, A2B, and A3) in all three commonly used prostate cancer cell lines, PC-3, DU145, and LNCaP. We explored the function of the A2B AR using PC-3 cells as a model. The A2B AR was visualized in PC-3 cells by laser confocal microscopy. The nonselective A2B AR agonist NECA and the selective A2B AR agonist BAY60-6583, but not the A2A AR agonist CGS21680, concentration-dependently induced adenosine 3′,5′-cyclic monophosphate (cyclic AMP) accumulation. NECA diminished lactate dehydrogenase (LDH) release, TNF-α-induced increase of caspase-3 activity, and cycloheximide (CHX)-induced morphological changes typical of apoptosis in PC-3 cells, which were blocked by a selective A2B AR antagonist PSB603. NECA-induced proliferation of PC-3 cells was diminished by siRNA specific for the A2B AR. The selective A2B AR antagonist PSB603 was shown to inhibit cell growth in all three cell lines. Thus, A2B AR blockade inhibits growth of prostate cancer cells, suggesting selective A2B AR antagonists as potential novel therapeutics.


Prostate cancer Cancer Adenosine receptor A2B G protein-coupled receptor (GPCR) Cell proliferation 



Bicinchoninic acid

Cyclic AMP

Adenosine 3′,5′-cyclic monophosphate








Dulbecco’s modified eagle’s medium




Fetal bovine serum


Glyceraldehyde-3-phosphate dehydrogenase


4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid


N 6-(3-Iodobenzyl)adenosine-5′-N-methyluronamide




Roswell Park Memorial Institute medium


Real time polymerase chain reaction


Sodium dodecyl sulfate polyacrylamide gel electrophoresis


2,3-bis(2-Methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt






Tumor necrosis factor-alpha



This study was supported by the NIDDK Intramural Research Program, National Institutes of Health, Bethesda, MD, USA; the National Natural Science Foundation of China (no.: 30940072); Guangdong Province Science and Technology Program (2012B031800263); and Nanfang Hospital, Southern Medical University, Guangzhou, China. The authors thank Dr. Yafang Hu (Children's National Medical Center, Washington, DC, USA) for assistance in confocal microscopy experiments. We thank Prof. Ad IJzerman (Leiden University, The Netherlands) for providing LUF6210 (BAY60-6583).

Conflicts of interest



  1. 1.
    Damber JE, Aus G (2008) Prostate Cancer 371:1710–1721Google Scholar
  2. 2.
    Lappano R, Maggiolini M (2011) G protein-coupled receptors: novel targets for drug discovery in cancer. Nat Rev Drug Discov 10:47–60PubMedCrossRefGoogle Scholar
  3. 3.
    Dorsam RT, Gutkind JS (2007) G-protein-coupled receptors and cancer. Nat Rev Cancer 7:79–94PubMedCrossRefGoogle Scholar
  4. 4.
    Wei Q, Costanzi S, Liu QZ, Gao ZG, Jacobson KA (2011) Activation of the P2Y1 receptor induces apoptosis and inhibits proliferation of prostate cancer cells. Biochem Pharmacol 82:418–425PubMedCrossRefGoogle Scholar
  5. 5.
    Li S, Huang S, Peng SB (2005) Overexpression of G protein-coupled receptors in cancer cells: involvement in tumor progression. Int J Oncol 27:1329–1339PubMedGoogle Scholar
  6. 6.
    Ma DF, Kondo T, Nakazawa T, Niu DF, Mochizuki K, Kawasaki T, Yamane T, Katoh R (2010) Hypoxia-inducible adenosine A2B receptor modulates proliferation of colon carcinoma cells. Hum Pathol 41:1550–1557PubMedCrossRefGoogle Scholar
  7. 7.
    Stagg J, Divisekera U, McLaughlin N, Sharkey J, Pommey S, Denoyer D, Dwyer KM, Smyth MJ (2010) Anti-CD73 antibody therapy inhibits breast tumor growth and metastasis. Proc Natl Acad Sci USA 107:1547–1552PubMedCrossRefGoogle Scholar
  8. 8.
    Ryzhov S, Novitskiy SV, Zaynagetdinov R, Goldstein AE, Carbone DP, Biaggioni I, Dikov MM, Feoktistov I (2008) Host A2B adenosine receptors promote carcinoma growth. Neoplasia 10:987–995PubMedGoogle Scholar
  9. 9.
    Cekic C, Sag D, Li Y, Theodorescu D, Strieter RM, Linden J (2012) Adenosine A2B receptor blockade slows growth of bladder and breast tumors. J Immunol 188:198–205PubMedCrossRefGoogle Scholar
  10. 10.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  11. 11.
    Blom WM, de Bont HJ, Meijerman I, Mulder GJ, Nagelkerke JF (1999) Prevention of cycloheximide-induced apoptosis in hepatocytes by adenosine and by caspase inhibitors. Biochem Pharmacol 58:1891–1898PubMedCrossRefGoogle Scholar
  12. 12.
    Das A, Durrant D, Mitchell C, Mayton E, Hoke NN, Salloum FN, Park MA, Qureshi I, Lee R, Dent P, Kukreja RC (2010) Sildenafil increases chemotherapeutic efficacy of doxorubicin in prostate cancer and ameliorates cardiac dysfunction. Proc Natl Acad Sci USA 107:18202–18207PubMedCrossRefGoogle Scholar
  13. 13.
    Janssens R, Boeynaems JM (2001) Effects of extracellular nucleotides and nucleosides on prostate carcinoma cells. Br J Pharmacol 132:536–546PubMedCrossRefGoogle Scholar
  14. 14.
    Aghaei M, Karami-Tehrani F, Panjehpour M, Salami S, Fallahian F (2011) Adenosine induces cell-cycle arrest and apoptosis in androgen-dependent and -independent prostate cancer cell lines, LNcap-FGC-10, DU-145, and PC3. Prostate 72:361–375PubMedCrossRefGoogle Scholar
  15. 15.
    Minelli A, Bellezza I, Tucci A, Rambotti MG, Conte C, Culig Z (2009) Differential involvement of reactive oxygen species and nucleoside transporters in cytotoxicity induced by two adenosine analogues in human prostate cancer cells. Prostate 69:538–547PubMedCrossRefGoogle Scholar
  16. 16.
    Panjehpour M, Movahedian A, Sadeghi H, Eghbali B, Yekdaneh A (2010) Adenosine receptor expression in two different human cancer lines at molecular level. Iranian J Cancer Prevention 3:111–116Google Scholar
  17. 17.
    Fishman P, Bar-Yehuda S, Ardon E, Rath-Wolfson L, Barrer F, Ochaion A, Madi L (2003) Targeting the A3 adenosine receptor for cancer therapy: inhibition of prostate carcinoma cell growth by A3AR agonist. Anticancer Res 23:2077–2083PubMedGoogle Scholar
  18. 18.
    Kalhan A, Gharibi B, Vazquez M, Jasani B, Neal J, Kidd M, Modlin IM, Pfragner R, Rees DA, Ham J (2012) Adenosine A2A and A2B receptor expression in neuroendocrine tumours: potential targets for therapy. Purinergic Signal 8:265–274PubMedCrossRefGoogle Scholar
  19. 19.
    Hastie C, Saxton M, Akpan A, Cramer R, Masters JR, Naaby-Hansen S (2005) Combined affinity labelling and mass spectrometry analysis of differential cell surface protein expression in normal and prostate cancer cells. Oncogene 24:5905–5913PubMedCrossRefGoogle Scholar
  20. 20.
    Stagg J, Beavis PA, Divisekera U, Liu MC, Moeller A, Darcy PK, Smyth MJ (2012) CD73-deficient mice are resistant to carcinogenesis. Cancer Res 72:2190–2196PubMedCrossRefGoogle Scholar
  21. 21.
    Feng L, Sun X, Csizmadia E, Han L, Bian S, Murakami T, Wang X, Robson SC, Wu Y (2011) Vascular CD39/ENTPD1 directly promotes tumor cell growth by scavenging extracellular adenosine triphosphate. Neoplasia 13:206–216PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht (outside the USA) 2013

Authors and Affiliations

  • Qiang Wei
    • 1
    • 2
  • Stefano Costanzi
    • 3
  • Ramachandran Balasubramanian
    • 1
  • Zhan-Guo Gao
    • 1
  • Kenneth A. Jacobson
    • 1
    Email author
  1. 1.Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA
  2. 2.Department of Urology, Nan Fang HospitalSouthern Medical UniversityGuangzhouPeople’s Republic of China
  3. 3.Laboratory of Biological Modeling, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA

Personalised recommendations