Skip to main content
SpringerLink
Log in

Characterization of neuroblastoma bone invasion/metastasis in established bone metastatic model of SY5Y and KCNR cell lines

  • Original Paper
  • Published:
Child's Nervous System Aims and scope Submit manuscript

Abstract

Objects

To determine the mechanism of neuroblastoma (NB) bone invasion/metastasis, it is necessary to investigate the bone invasion/metastasis-related factors in the bone invasion/metastasis process. Some evidence has suggested that various proteins were involved in bone osteolytic response. The invasion/metastasis property and gene expression of NB, however, are still unknown.

Methods

Single-cell suspensions of SY5Y and KCNR cells were injected directly into the femur of nude mice. Radiological and histological analyses, immunohistochemistry analyses, and western blot assay were performed to characterize bone metastasis mechanism in these bone metastasis models.

Results

SY5Y and KCNR NB cells result in osteolytic responses in bone metastasis model. Osteoprotegerin (OPG), receptor activator of NF-kappaB ligand (RANKL), parathyroid hormone-related peptide (PTHrP), endothelin 1 (ET-1), and CXCR4 were examined and compared among in vitro, in vivo, and normal bone, respectively. PTHrP, OPG, RANKL, and ET-1 except CXCR4 in SY5Y and KCNR NB cells xenografts were strikingly upregulated compared with normal bone and NB cells. However, significantly stronger expression of PTHrP and RANKL was presented than ET-1 and OPG; furthermore, the ratios of expression of PTHrP, RANKL to OPG, and ET-1 were also markedly increased in vivo versus in vitro.

Conclusions

Our study provided evidence that NB cell may enhance bone invasion through PTHrP, OPG, RANKL, and ET-1, especially PTHrP and RANKL which may display stronger effects. CXCR4 appeared not participating in bone invasion, but in tumor growth, and homing to bone. Targeting PTHrP, OPG, ET-1, and RANKL may provide a new insight and method for patient therapy by inhibiting NB bone metastasis and invasiveness.

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

Similar content being viewed by others

References

  1. Ara T, DeClerck YA (2006) Mechanisms of invasion and metastasis in human neuroblastoma. Cancer Metastasis Rev 25:645–657

    Article  PubMed  Google Scholar 

  2. Armstrong JL, Martin S, Illingworth NA, Jamieson D, Neilson A, Lovat PE, Redfern CP, Veal GJ (2012) The impact of retinoic acid treatment on the sensitivity of neuroblastoma cells to fenretinide. Oncol Rep 27:293–298

    CAS  PubMed  Google Scholar 

  3. Bilir A, Erguven M, Yazihan N, Aktas E, Oktem G, Sabanci A (2010) Enhancement of vinorelbine-induced cytotoxicity and apoptosis by clomipramine and lithium chloride in human neuroblastoma cancer cell line SH-SY5Y. J Neurooncol 100:385–395

    Article  CAS  PubMed  Google Scholar 

  4. Brodeur GM (2003) Neuroblastoma: biological insights into a clinical enigma. Nat Rev Cancer 3:203–216

    Article  CAS  PubMed  Google Scholar 

  5. Carlisle AJ, Lyttle CA, Carlisle RY, Maris JM (2009) CXCR4 expression heterogeneity in neuroblastoma cells due to ligand-independent regulation. Mol Canc 8:126

    Article  Google Scholar 

  6. Chappard D, Bouvard B, Baslé MF, Legrand E, Audran M (2011) Bone metastasis: histological changes and pathophysiological mechanisms in osteolytic or osteosclerotic localizations. A review. Morphologie 95:65–75

    Article  CAS  PubMed  Google Scholar 

  7. Chawla M, Kumar R, Agarwala S, Bakhshi S, Gupta DK, Malhotra (2010) A Role of positron emission tomography-computed tomography in staging and early chemotherapy response evaluation in children with neuroblastoma. Indian J Nucl Med 25:147–155

    PubMed  Google Scholar 

  8. Davidoff AM (2012) Neuroblastoma. Semin Pediatr Surg 21:2–14

    Article  PubMed  Google Scholar 

  9. Dickey A, Schleicher S, Leahy K, Hu R, Hallahan D, Thotala DK (2011) GSK-3β inhibition promotes cell death, apoptosis, and in vivo tumor growth delay in neuroblastoma Neuro-2A cell line. J Neurooncol 104:145–153

    Article  CAS  PubMed  Google Scholar 

  10. Enjuanes A, Ruiz-Gaspà S, Peris P, Ozalla D, Álvarez L, Combalia A, Martínez de Osaba MJ, Monegal A, Pares A, Guañabens N (2010) The effect of the alendronate on OPG/RANKL system in differentiated primary human osteoblasts. Endocrine 37:180–186

    Article  CAS  PubMed  Google Scholar 

  11. Granchi D, Corrias MV, Garaventa A, Baglìo SR, Cangemi G, Carlini B, Paolucci P, Giunti A, Baldini N (2011) Neuroblastoma and bone metastases: clinical significance and prognostic value of Dickkopf 1 plasma levels. Bone 48:152–159

    Article  CAS  PubMed  Google Scholar 

  12. Gulenchyn KY, Yao X, Asa SL, Singh S, Law C (2012) Radionuclide therapy in neuroendocrine tumours: a systematic review. Clin Oncol (R Coll Radiol) 24:294–308

    Article  CAS  Google Scholar 

  13. Ishola TA, Chung DH (2007) Neuroblastoma. Surg Oncol 16:149–156

    Article  PubMed  Google Scholar 

  14. Jabbar S, Drury J, Fordham JN, Datta HK, Francis RM, Tuck SP (2011) Osteoprotegerin, RANKL and bone turnover in postmenopausal osteoporosis. J Clin Pathol 64:354–357

    Article  CAS  PubMed  Google Scholar 

  15. Klymenko T, Brandenburg M, Morrow C, Dive C, Makin G (2011) The novel Bcl-2 inhibitor ABT-737 is more effective in hypoxia and is able to reverse hypoxia-induced drug resistance in neuroblastoma cells. Mol Cancer Ther 10:2373–2383

    Article  CAS  PubMed  Google Scholar 

  16. Ma M, Ye JY, Deng R, Dee CM, Chan GC (2011) Mesenchymal stromal cells may enhance metastasis of neuroblastoma via SDF-1/CXCR4 and SDF-1/CXCR7 signaling. Cancer Lett 312:1–10

    Article  CAS  PubMed  Google Scholar 

  17. Maris JM, Hogarty MD, Bagatell R, Cohn SL (2007) Neuroblastoma. Lancet 369:2106–2120

    Article  CAS  PubMed  Google Scholar 

  18. Matthay KK, Shulkin B, Ladenstein R, Michon J, Giammarile F, Lewington V, Pearson AD, Cohn SL (2010) Criteria for evaluation of disease extent by (123)I-metaiodobenzylguanidine scans in neuroblastoma: a report for the International Neuroblastoma Risk Group (INRG) Task Force. Br J Cancer 102:1319–1326

    Article  CAS  PubMed  Google Scholar 

  19. Meier R, Mühlethaler-Mottet A, Flahaut M, Coulon A, Fusco C, Louache F, Auderset K, Bourloud KB, Daudigeos E, Ruegg C, Vassal G, Gross N, Joseph JM (2007) The chemokine receptor CXCR4 strongly promotes neuroblastoma primary tumour and metastatic growth, but not invasion. PLoS One 2:e1016

    Article  PubMed  Google Scholar 

  20. Ostrowska Z, Ziora K, Oświęcimska J, Swiętochowska E, Szapska B, Wołkowska-Pokrywa K, Dyduch A (2012) RANKL/RANK/OPG system and bone status in females with anorexia nervosa. Bone 50:156–160

    Article  CAS  PubMed  Google Scholar 

  21. Park JR, Eggert A, Caron H (2008) Neuroblastoma: biology, prognosis, and treatment. Pediatr Clin North Am 55:97–120

    Article  PubMed  Google Scholar 

  22. Park JR, Scott JR, Stewart CF, London WB, Naranjo A, Santana VM, Shaw PJ, Cohn SL, Matthay KK (2011) Pilot induction regimen incorporating pharmacokinetically guided topotecan for treatment of newly diagnosed high-risk neuroblastoma: a Children's Oncology Group study. J Clin Oncol 29:4351–4357

    Article  CAS  PubMed  Google Scholar 

  23. Peng H, Sohara Y, Moats RA, Nelson MD Jr, Groshen SG, Ye W, Reynolds CP, DeClerck YA (2007) The activity of zoledronic acid on neuroblastoma bone metastasis involves inhibition of osteoclasts and tumor cell survival and proliferation. Cancer Res 67(19):9346–9355

    Article  CAS  PubMed  Google Scholar 

  24. Poretti A, Grotzer MA (2012) Neuroblastoma with spinal cord compression: is there an emergency treatment of choice? Dev Med Child Neurol 54:297–298

    Article  PubMed  Google Scholar 

  25. Roodman GD (2004) Mechanisms of bone metastasis. N Engl J Med 350:655–664

    Article  Google Scholar 

  26. Sartelet H, Durrieu L, Fontaine F, Nyalendo C, Haddad E (2012) Description of a new xenograft model of metastatic neuroblastoma using NOD/SCID/Il2rg null (NSG) mice. In Vivo 26:19–29

    PubMed  Google Scholar 

  27. Seeger RC (2011) Immunology and immunotherapy of neuroblastoma. Semin Cancer Biol 21:229–237

    Article  CAS  PubMed  Google Scholar 

  28. Sharp SE, Gelfand MJ, Shulkin BL (2011) Pediatrics: diagnosis of neuroblastoma. Semin Nucl Med 41:345–353

    Article  PubMed  Google Scholar 

  29. Sohara Y, Shimada H, DeClerck YA (2005) Mechanisms of bone invasion and metastasis in human neuroblastoma. Cancer Lett 228:203–209

    Article  CAS  PubMed  Google Scholar 

  30. Sohara Y, Shimada H, Scadeng M, Pollack H, Yamada S, Ye W, Reynolds CP, DeClerck YA (2003) Lytic bone lesions in human neuroblastoma xenograft involve osteoclast recruitment and are inhibited by bisphosphonate. Cancer Res 63:3026–3031

    CAS  PubMed  Google Scholar 

  31. Suva LJ, Washam C, Nicholas RW, Griffin RJ (2011) Bone metastasis: mechanisms and therapeutic opportunities. Nat Rev Endocrinol 7:208–218

    Article  CAS  PubMed  Google Scholar 

  32. Taggart DR, London WB, Schmidt ML, DuBois SG, Monclair TF, Nakagawara A, De Bernardi B, Ambros PF, Pearson AD, Cohn SL, Matthay KK (2011) Prognostic value of the stage 4S metastatic pattern and tumor biology in patients with metastatic neuroblastoma diagnosed between birth and 18 months of age. J Clin Oncol 29:4358–4364

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by National Natural Science Foundation grant from the China National Science Foundation Committee (project code 81172410).

Conflict of interest

The authors have no conflict to report.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Shengjing Hospital, China Medical University, Shenyang, China

    Hongyu Zhao, Shuai Li, Zuke Da, Liang Ma, Yaoxin Lin & Debao Zhi

  2. Department of Oncology, Shengjing Hospital, China Medical University, Shenyang, China

    Weisong Cai

  3. Department of Pathology, Shengjing Hospital, China Medical University, Shenyang, China

    Hanxue Sun

Authors
  1. Hongyu Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Weisong Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shuai Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zuke Da
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hanxue Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Liang Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yaoxin Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Debao Zhi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hongyu Zhao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, H., Cai, W., Li, S. et al. Characterization of neuroblastoma bone invasion/metastasis in established bone metastatic model of SY5Y and KCNR cell lines. Childs Nerv Syst 29, 1097–1105 (2013). https://doi.org/10.1007/s00381-013-2086-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00381-013-2086-8

Keywords

Search

Navigation