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Differential impact of adenosine nucleotides released by osteocytes on breast cancer growth and bone metastasis

Oncogene volume 34, pages 1831–1842 (2015)Cite this article

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Abstract

Extracellular ATP has been shown to either inhibit or promote cancer growth and migration; however, the mechanism underlying this discrepancy remained elusive. Here we demonstrate the divergent roles of ATP and adenosine released by bone osteocytes on breast cancers. We showed that conditioned media (CM) collected from osteocytes treated with alendronate (AD), a bisphosphonate drug, inhibited the migration of human breast cancer MDA-MB-231 cells. Removal of the extracellular ATP by apyrase in CM abolished this effect, suggesting the involvement of ATP. ATP exerted its inhibitory effect through the activation of purinergic P2X receptor signaling in breast cancer cells evidenced by the attenuation of the inhibition by an antagonist, oxidized ATP, as well as knocking down P2X7 with small interfering RNA (siRNA), and the inhibition of migration by an agonist, BzATP. Intriguingly, ATP had a biphasic effect on breast cancer cells—lower dosage inhibited but higher dosage promoted its migration. The stimulatory effect on migration was blocked by an adenosine receptor antagonist, MRS1754, ARL67156, an ecto-ATPase inhibitor, and A2A receptor siRNA, suggesting that in contrast to ATP, adenosine, a metabolic product of ATP, promoted migration of breast cancer cells. Consistently, non-hydrolyzable ATP, ATPγS, only inhibited but did not promote cancer cell migration. ATP also had a similar inhibitory effect on the Py8119 mouse mammary carcinoma cells; however, adenosine had no effect owing to the absence of the A2A receptor. Consistently, ATPγS inhibited, whereas adenosine promoted anchorage-independent growth of MDA-MB-231 cells. Our in vivo xenograft study showed a significant delay of tumor growth with the treatment of ATPγS. Moreover, the extent of bone metastasis in a mouse intratibial model was significantly reduced with the treatment of ATPγS. Together, our results suggest the distinct roles of ATP and adenosine released by osteocytes and the activation of corresponding receptors P2X7 and A2A signaling on breast cancer cell growth, migration and bone metastasis.

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References

  1. Kohno N . Treatment of breast cancer with bone metastasis: bisphosphonate treatment - current and future. Int J Clin Oncol 2008; 13: 18–23.

    Article  CAS  PubMed  Google Scholar 

  2. Jin JK, Dayyani F, Gallick GE . Steps in prostate cancer progression that lead to bone metastasis. Int J Cancer 2011; 128: 2545–2561.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Welch DR, Harms JF, Mastro AM, Gay CV, Donahue HJ . Breast cancer metastasis to bone: evolving models and research challenges. J Musculoskelet Neuronal Interact 2003; 3: 30–38.

    CAS  PubMed  Google Scholar 

  4. Roodman GD . Mechanism of bone metastasis. New Engl J Med 2004; 350: 1655–1664.

    Article  CAS  PubMed  Google Scholar 

  5. van der Pluijm G, Sijmons B, Vloedgraven H, Deckers M, Papapoulos S, Lowik C . Monitoring metastatic behavior of human tumor cells in mice with species-specific polymerase chain reaction: elevated expression of angiogenesis and bone resporption stimulators by breast cancer in bone metastases. J Bone Miner Res 2001; 16: 1077–1091.

    Article  CAS  PubMed  Google Scholar 

  6. Bonewald LF . Osteocytes as dynamic multifucntional cells. Ann NY Acad Sci 2007; 1116: 281–290.

    Article  CAS  PubMed  Google Scholar 

  7. Matsuo K . Cross-talk among bone cells. Curr Opin Nephrol Hypertens 2009; 18: 292–297.

    Article  CAS  PubMed  Google Scholar 

  8. Brown SA, Guise TA . Cancer-associated bone disease. Curr Osteopor Rep 2007; 5: 120–127.

    Article  Google Scholar 

  9. Plotkin LI, Manolagas SC, Bellido T . Transduction of cell survival signals by connexin-43 hemichannels. J Biol Chem 2002; 277: 8648–8657.

    Article  CAS  PubMed  Google Scholar 

  10. Genetos DC, Kephart CJ, Zhang Y, Yellowley CE, Donahue HJ . Oscillating fluid flow activation of gap junction hemichannels induces ATP release from MLO-Y4 osteocytes. J Cell Physiol 2007; 212: 207–214.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Shabbir M, Burnstock G . Purinergic receptor-mediated effects of adenosine 5′-triphosphate in urological malignant diseases. Int J Urol 2009; 16: 143–150.

    Article  CAS  PubMed  Google Scholar 

  12. White N, Burnstock G . P2 receptors and cancer. Trends Pharmacol Sci 2006; 27: 211–217.

    Article  CAS  PubMed  Google Scholar 

  13. Rapaport E, Fishman RF, Gercel C . Growth inhibition of human tumor cells in soft-agar cultures by treatment with low levels of adenosine 5′-triphosphate. Cancer Res 1983; 43: 4402–4406.

    CAS  PubMed  Google Scholar 

  14. Salvestrini V, Zini R, Rossi L, Gulinelli S, Manfredini R, Bianchi E et al. Purinergic signaling inhibits human acute myeloblastic leukemia cell proliferation, migration, and engraftment in immunodeficient mice. Blood 2012; 119: 217–226.

    Article  CAS  PubMed  Google Scholar 

  15. Rapaport E . Experimental cancer therapy in mice by adenine nucleotides. Eur J Cancer Clin Oncol 1988; 24: 1491–1497.

    Article  CAS  PubMed  Google Scholar 

  16. Deli T, Csernoch L . Extracellular ATP and cancer: an overview with special reference to P2 purinergic receptors. Pathol Oncol Res 2008; 14: 219–231.

    Article  CAS  PubMed  Google Scholar 

  17. Savariar EN, Felsen CN, Nashi N, Jiang T, Ellies LG, Steinbach P et al. Real-time in vivo molecular detection of primary tumors and metastases with ratiometric activatable cell-penetrating peptides. Cancer Res 2013; 73: 855–864.

    Article  CAS  PubMed  Google Scholar 

  18. Abbracchio MP, Burnstock G, Boeynaems JM, Barnard EA, Boyer JL, Kennedy C et al. International Union of Pharmacology LVIII: update on the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev 2006; 58: 281–341.

    Article  CAS  PubMed  Google Scholar 

  19. Batra N, Kar R, Jiang JX . Gap junctions and hemichannels in signal transmission, function and development of bone. Biochim Biophys Acta 2012; 1818: 1909–1918.

    Article  CAS  PubMed  Google Scholar 

  20. Burnstock G . Unresolved issues and controversites in purinergic signalling. J Physiol 2008; 586: 3307–3312.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Ohta A, Gorelik E, Prasad SJ, Ronchese F, Lukashev D, Wong MK et al. A2A adenosine receptor protects tumors from antitumor T cells. Proc Natl Acad Sci USA 2006; 103: 13132–13137.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Falzoni S, Donvito GD, Di Virgilio F . Detecting adenosine triphosphate in the pericellular space. Interface Focus 2013; 3: 20120101.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Leth-Larsen R, Lund R, Hansen HV, Laenkholm AV, Tarin D, Jensen ON et al. Metastasis-related plasma membrane proteins of human breast cancer cells identified by comparative quantitative mass spectrometry. Mol Cell Proteomics 2009; 8: 1436–1449.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Zhi X, Chen S, Zhou P, Shao Z, Wang L, Ou Z et al. RNA interference of ecto-5′-nucleotidase (CD73) inhibits human breast cancer cell growth and invasion. Clin Exp Metastasis 2007; 24: 439–448.

    Article  CAS  PubMed  Google Scholar 

  25. Wang L, Zhou X, Zhou T, Ma D, Chen S, Zhi X et al. Ecto-5′-nucleotidase promotes invasion, migration and adhesion of human breast cancer cells. J Cancer Res Clin Oncol 2008; 134: 365–372.

    Article  CAS  PubMed  Google Scholar 

  26. Jelassi B, Chantôme A, Alcaraz-Pérez F, Baroja-Mazo A, Cayuela ML, Pelegrin P et al. P2X(7) receptor activation enhances SK3 channels- and cystein cathepsin-dependent cancer cells invasiveness. Oncogene 2011; 30: 2108–2122.

    Article  CAS  PubMed  Google Scholar 

  27. Woodhouse EC, Amanatullah DF, Schetz JA, Liotta LA, Stracke ML, Clair T . Adenosine receptor mediates motility in human melanoma cells. Biochem Biophys Res Commun 1998; 246: 888–894.

    Article  CAS  PubMed  Google Scholar 

  28. Gessi S, Merighi S, Sacchetto V, Simioni C, Borea PA . Adenosine receptors and cancer. Biochim Biophys Acta 2011; 1808: 1400–1412.

    Article  CAS  PubMed  Google Scholar 

  29. Campbell JP, Merkel AR, Masood-Campbell SK, Elefteriou F, Sterling JA . Models of bone metastasis. J Vis Exp 2012; 67: e4260.

    Google Scholar 

  30. Kretschmann KL, Welm AL . Mouse models of breast cancer metastasis to bone. Cancer Metastasis Rev 2012; 31: 579–583.

    Article  PubMed  Google Scholar 

  31. Agteresch HJ, Dagnelie PC, Rietveld T, van den Berg JW, Danser AH, Wilson JH . Pharmacokinetics of intravenous ATP in cancer patients. Eur J Clin Pharmacol 2000; 56: 49–55.

    Article  CAS  PubMed  Google Scholar 

  32. Agteresch HJ, Burgers SA, van der GA, Wilson JH, Dagnelie PC . Randomized clinical trial of adenosine 5′-triphosphate on tumor growth and survival in advanced lung cancer patients. Anticancer Drugs 2003; 14: 639–644.

    Article  CAS  PubMed  Google Scholar 

  33. Beijer S, Hupperets PS, van den Borne BE, Eussen SR, van Henten AM, van den Beuken-van Everdingen M et al. Effect of adenosine 5′-triphosphate infusions on the nutritional status and survival of preterminal cancer patients. Anticancer Drugs 2009; 20: 625–633.

    Article  CAS  PubMed  Google Scholar 

  34. Zhang W, Tan S, Paintsil E, Dutschman GE, Gullen EA, Chu E et al. Analysis of deoxyribonucleotide pools in human cancer cell lines using a liquid chromatography coupled with tandem mass spectrometry technique. Biochem Pharmacol 2011; 82: 411–417.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Mishra S, Tang Y, Wang L, DeGraffenried L, Yeh IT, Werner S et al. Blockade of transforming growth factor-beta (TGFbeta) signaling inhibits osteoblastic tumorigenesis by a novel human prostate cancer cell line. Prostate 2011; 71: 1441–1454.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Dr Sumin Gu, Dr Sweta Mishra and Dr Tanuka Biswas for technical assistance. The work was supported by the Welch Foundation grant AQ-1507 and National Institutes of Health grant EY012085 to JXJ, ES022057 to L-ZS, CA118182 to LGE and the NCI Cancer Center Support grant 2 P30 CA054174-17 to Cancer Therapy and Research Center.

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Authors and Affiliations

  1. Department of Biochemistry, University of Texas Health Science Center, San Antonio, TX, USA

    J Z Zhou, M A Riquelme, X Gao & J X Jiang

  2. Department of Pathology, University of California, San Diego, La Jolla, CA, USA

    L G Ellies

  3. Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio, TX, USA

    L Z Sun

  4. Cancer Therapy and Research Center, University of Texas Health Science Center, San Antonio, TX, USA

    L Z Sun

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  1. J Z Zhou
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Correspondence to J X Jiang.

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Zhou, J., Riquelme, M., Gao, X. et al. Differential impact of adenosine nucleotides released by osteocytes on breast cancer growth and bone metastasis. Oncogene 34, 1831–1842 (2015). https://doi.org/10.1038/onc.2014.113

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