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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. Jacobson
Original Article

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.

Keywords

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

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

Notes

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.

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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
  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

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