Skip to main content
SpringerLink
Log in

A2B adenosine receptor blockade inhibits growth of prostate cancer cells

  • Original Article
  • Published:
Purinergic Signalling Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

BCA:

Bicinchoninic acid

Cyclic AMP:

Adenosine 3′,5′-cyclic monophosphate

CGS21680:

(2-[p-(2-Carboxyethyl)phenylethylamino]-5′-N-ethylcarboxamidoadenosine)

CHX:

Cycloheximide

DAPI:

4′,6-diamidino-2-phenylindole

DMEM:

Dulbecco’s modified eagle’s medium

DOX:

Doxorubicin

FBS:

Fetal bovine serum

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HEPES:

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

IB-MECA:

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

MRS2365:

(N)-Methanocarba-2′-deoxy-2-methylthio-adenosine-5′-diphosphate

RPMI:

Roswell Park Memorial Institute medium

RT-PCR:

Real time polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

XTT:

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

NECA:

5′-N-Ethylcarboxamidoadenosine

PSB603:

8-[4-[4-(4-Chlorophenzyl)piperazide-1-sulfonyl)phenyl]]-1-propylxanthine

TNF-α:

Tumor necrosis factor-alpha

References

  1. Damber JE, Aus G (2008) Prostate Cancer 371:1710–1721

    Google Scholar 

  2. Lappano R, Maggiolini M (2011) G protein-coupled receptors: novel targets for drug discovery in cancer. Nat Rev Drug Discov 10:47–60

    Article  PubMed  CAS  Google Scholar 

  3. Dorsam RT, Gutkind JS (2007) G-protein-coupled receptors and cancer. Nat Rev Cancer 7:79–94

    Article  PubMed  CAS  Google Scholar 

  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 82:418–425

    Article  PubMed  CAS  Google Scholar 

  5. Li S, Huang S, Peng SB (2005) Overexpression of G protein-coupled receptors in cancer cells: involvement in tumor progression. Int J Oncol 27:1329–1339

    PubMed  CAS  Google Scholar 

  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 41:1550–1557

    Article  PubMed  CAS  Google Scholar 

  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 107:1547–1552

    Article  PubMed  CAS  Google Scholar 

  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 10:987–995

    PubMed  CAS  Google Scholar 

  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 188:198–205

    Article  PubMed  CAS  Google Scholar 

  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 25:402–408

    Article  PubMed  CAS  Google Scholar 

  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 58:1891–1898

    Article  PubMed  CAS  Google Scholar 

  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 107:18202–18207

    Article  PubMed  CAS  Google Scholar 

  13. Janssens R, Boeynaems JM (2001) Effects of extracellular nucleotides and nucleosides on prostate carcinoma cells. Br J Pharmacol 132:536–546

    Article  PubMed  CAS  Google Scholar 

  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 72:361–375

    Article  PubMed  Google Scholar 

  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 69:538–547

    Article  PubMed  CAS  Google Scholar 

  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 3:111–116

    CAS  Google Scholar 

  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 23:2077–2083

    PubMed  CAS  Google Scholar 

  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 8:265–274

    Article  PubMed  CAS  Google Scholar 

  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 24:5905–5913

    Article  PubMed  CAS  Google Scholar 

  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 72:2190–2196

    Article  PubMed  CAS  Google Scholar 

  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 13:206–216

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

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

None.

Author information

Author notes

  1. Stefano Costanzi

    Present address: Department of Chemistry, American University, Washington, DC, 20016, USA

Authors and Affiliations

  1. Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892-0810, USA

    Qiang Wei, Ramachandran Balasubramanian, Zhan-Guo Gao & Kenneth A. Jacobson

  2. Department of Urology, Nan Fang Hospital, Southern Medical University, Guangzhou, 510515, People’s Republic of China

    Qiang Wei

  3. Laboratory of Biological Modeling, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892-0810, USA

    Stefano Costanzi

Authors
  1. Qiang Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stefano Costanzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ramachandran Balasubramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhan-Guo Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kenneth A. Jacobson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kenneth A. Jacobson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wei, Q., Costanzi, S., Balasubramanian, R. et al. A2B adenosine receptor blockade inhibits growth of prostate cancer cells. Purinergic Signalling 9, 271–280 (2013). https://doi.org/10.1007/s11302-012-9350-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11302-012-9350-3

Keywords

Search

Navigation