Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

BRG1/SMARCA4 is essential for neuroblastoma cell viability through modulation of cell death and survival pathways

Oncogene volume 35, pages 5179–5190 (2016)Cite this article

Subjects

Abstract

Neuroblastoma (NB) is a neoplasm of the sympathetic nervous system, and is the most common solid tumor of infancy. NBs are very heterogeneous, with a clinical course ranging from spontaneous regression to resistance to all current forms of treatment. High-risk patients need intense chemotherapy, and only 30–40% will be cured. Relapsed or metastatic tumors acquire multi-drug resistance, raising the need for alternative treatments. Owing to the diverse mechanisms that are responsible of NB chemoresistance, we aimed to target epigenetic factors that control multiple pathways to bypass therapy resistance. We found that the SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 4 (SMARCA4/BRG1) was consistently upregulated in advanced stages of NB, with high BRG1 levels being indicative of poor outcome. Loss-of-function experiments in vitro and in vivo showed that BRG1 is essential for the proliferation of NB cells. Furthermore, whole-genome transcriptome analysis revealed that BRG1 controls the expression of key elements of oncogenic pathways such as PI3K/AKT and BCL2, which offers a promising new combination therapy for high-risk NB.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

Abbreviations

BET:

bromodomain and extraterminal domain

MYCN:

V-Myc Avian Myelocytomatosis Viral Oncogene Neuroblastoma Derived Homolog

NB:

neuroblastoma

NSC:

non-silencing control

PI3K:

phosphatidylinositol-4,5-bisphosphate 3-kinase

qPCR:

quantitative real-time PCR

SWI/SNF:

SWItch/Sucrose NonFermentable.

References

  1. Yu AL, Gilman AL, Ozkaynak MF, London WB, Kreissman SG, Chen HX et al. Anti-GD2 antibody with GM-CSF, interleukin-2, and isotretinoin for neuroblastoma. N Engl J Med 2010; 363: 1324–1334.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Irwin MS, Park JR . Neuroblastoma: paradigm for precision medicine. Pediatr Clin North Am 2015; 62: 225–256.

    Article  PubMed  Google Scholar 

  3. Burke MJ, Lamba JK, Pounds S, Cao X, Ghodke-Puranik Y, Lindgren BR et al. A therapeutic trial of decitabine and vorinostat in combination with chemotherapy for relapsed/refractory acute lymphoblastic leukemia. Am J Hematol 2014; 89: 889–895.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Popovic R, Licht JD . Emerging epigenetic targets and therapies in cancer medicine. Cancer Discov 2012; 2: 405–413.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Delmore JE, Issa GC, Lemieux ME, Rahl PB, Shi J, Jacobs HM et al. BET bromodomain inhibition as a therapeutic strategy to target c-Myc. Cell 2011; 146: 904–917.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Cheng Z, Gong Y, Ma Y, Lu K, Lu X, Pierce LA et al. Inhibition of BET bromodomain targets genetically diverse glioblastoma. Clin Cancer Res 2013; 19: 1748–1759.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Segura MF, Fontanals-Cirera B, Gaziel-Sovran A, Guijarro MV, Hanniford D, Zhang G et al. BRD4 sustains proliferation and represents a new target for epigenetic therapy in melanoma. Cancer Res 2013; 73: 6264–6276.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Puissant A, Frumm SM, Alexe G, Bassil CF, Qi J, Chanthery YH et al. Targeting MYCN in neuroblastoma by BET bromodomain inhibition. Cancer Discov 2013; 3: 308–323.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Wyce A, Ganji G, Smitheman KN, Chung CW, Korenchuk S, Bai Y et al. BET inhibition silences expression of MYCN and BCL2 and induces cytotoxicity in neuroblastoma tumor models. PLoS One 2013; 8: e72967.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Shi J, Whyte WA, Zepeda-Mendoza CJ, Milazzo JP, Shen C, Roe JS et al. Role of SWI/SNF in acute leukemia maintenance and enhancer-mediated Myc regulation. Genes Dev 2013; 27: 2648–2662.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Shain AH, Pollack JR . The spectrum of SWI/SNF mutations, ubiquitous in human cancers. PLoS One 2013; 8: e55119.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Cote J, Quinn J, Workman JL, Peterson CL . Stimulation of GAL4 derivative binding to nucleosomal DNA by the yeast SWI/SNF complex. Science 1994; 265: 53–60.

    Article  CAS  PubMed  Google Scholar 

  13. Kadam S, Emerson BM . Transcriptional specificity of human SWI/SNF BRG1 and BRM chromatin remodeling complexes. Mol Cell 2003; 11: 377–389.

    Article  CAS  PubMed  Google Scholar 

  14. Sentani K, Oue N, Kondo H, Kuraoka K, Motoshita J, Ito R et al. Increased expression but not genetic alteration of BRG1, a component of the SWI/SNF complex, is associated with the advanced stage of human gastric carcinomas. Pathobiology 2001; 69: 315–320.

    Article  CAS  PubMed  Google Scholar 

  15. Li Y, Shi QL, Jin XZ, Meng K, Zhou XJ, Sun LP . BRG1 expression in prostate carcinoma by application of tissue microarray. Zhonghua Nan Ke Xue 2006; 12: 629–632.

    CAS  PubMed  Google Scholar 

  16. Sun A, Tawfik O, Gayed B, Thrasher JB, Hoestje S, Li C et al. Aberrant expression of SWI/SNF catalytic subunits BRG1/BRM is associated with tumor development and increased invasiveness in prostate cancers. Prostate 2007; 67: 203–213.

    Article  CAS  PubMed  Google Scholar 

  17. Lin H, Wong RP, Martinka M, Li G . BRG1 expression is increased in human cutaneous melanoma. Br J Dermatol 2010; 163: 502–510.

    Article  CAS  PubMed  Google Scholar 

  18. Watanabe T, Semba S, Yokozaki H . Regulation of PTEN expression by the SWI/SNF chromatin-remodelling protein BRG1 in human colorectal carcinoma cells. Br J Cancer 2011; 104: 146–154.

    Article  CAS  PubMed  Google Scholar 

  19. Bai J, Mei PJ, Liu H, Li C, Li W, Wu YP et al. BRG1 expression is increased in human glioma and controls glioma cell proliferation, migration and invasion in vitro. J Cancer Res Clin Oncol 2012; 138: 991–998.

    Article  CAS  PubMed  Google Scholar 

  20. Numata M, Morinaga S, Watanabe T, Tamagawa H, Yamamoto N, Shiozawa M et al. The clinical significance of SWI/SNF complex in pancreatic cancer. Int J Oncol 2013; 42: 403–410.

    Article  PubMed  Google Scholar 

  21. Bai J, Mei P, Zhang C, Chen F, Li C, Pan Z et al. BRG1 is a prognostic marker and potential therapeutic target in human breast cancer. PLoS One 2013; 8: e59772.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Buscarlet M, Krasteva V, Ho L, Simon C, Hebert J, Wilhelm B et al. Essential role of BRG, the ATPase subunit of BAF chromatin remodeling complexes, in leukemia maintenance. Blood 2014; 123: 1720–1728.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Romero OA, Setien F, John S, Gimenez-Xavier P, Gomez-Lopez G, Pisano D et al. The tumour suppressor and chromatin-remodelling factor BRG1 antagonizes Myc activity and promotes cell differentiation in human cancer. EMBO Mol Med 2012; 4: 603–616.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Fan QW, Knight ZA, Goldenberg DD, Yu W, Mostov KE, Stokoe D et al. A dual PI3 kinase/mTOR inhibitor reveals emergent efficacy in glioma. Cancer Cell 2006; 9: 341–349.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Souers AJ, Leverson JD, Boghaert ER, Ackler SL, Catron ND, Chen J et al. ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat Med 2013; 19: 202–208.

    Article  CAS  PubMed  Google Scholar 

  26. Tse C, Shoemaker AR, Adickes J, Anderson MG, Chen J, Jin S et al. ABT-263: a potent and orally bioavailable Bcl-2 family inhibitor. Cancer Res 2008; 68: 3421–3428.

    Article  CAS  PubMed  Google Scholar 

  27. Kumar HR, Zhong X, Hoelz DJ, Rescorla FJ, Hickey RJ, Malkas LH et al. Three-dimensional neuroblastoma cell culture: proteomic analysis between monolayer and multicellular tumor spheroids. Pediatr Surg Int 2008; 24: 1229–1234.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Yang Q, Kiernan CM, Tian Y, Salwen HR, Chlenski A, Brumback BA et al. Methylation of CASP8, DCR2, and HIN-1 in neuroblastoma is associated with poor outcome. Clin Cancer Res 2007; 13: 3191–3197.

    Article  CAS  PubMed  Google Scholar 

  29. Buckley PG, Das S, Bryan K, Watters KM, Alcock L, Koster J et al. Genome-wide DNA methylation analysis of neuroblastic tumors reveals clinically relevant epigenetic events and large-scale epigenomic alterations localized to telomeric regions. Int J Cancer 2011; 128: 2296–2305.

    Article  CAS  PubMed  Google Scholar 

  30. Alaminos M, Davalos V, Cheung NK, Gerald WL, Esteller M . Clustering of gene hypermethylation associated with clinical risk groups in neuroblastoma. J Natl Cancer Inst 2004; 96: 1208–1219.

    Article  CAS  PubMed  Google Scholar 

  31. Weber M, Davies JJ, Wittig D, Oakeley EJ, Haase M, Lam WL et al. Chromosome-wide and promoter-specific analyses identify sites of differential DNA methylation in normal and transformed human cells. Nat Genet 2005; 37: 853–862.

    Article  CAS  PubMed  Google Scholar 

  32. Grau E, Martinez F, Orellana C, Canete A, Yanez Y, Oltra S et al. Epigenetic alterations in disseminated neuroblastoma tumour cells: influence of TMS1 gene hypermethylation in relapse risk in NB patients. J Cancer Res Clin Oncol 2010; 136: 1415–1421.

    Article  CAS  PubMed  Google Scholar 

  33. Charlet J, Schnekenburger M, Brown KW, Diederich M . DNA demethylation increases sensitivity of neuroblastoma cells to chemotherapeutic drugs. Biochem Pharmacol 2012; 83: 858–865.

    Article  CAS  PubMed  Google Scholar 

  34. Fouladi M, Park JR, Stewart CF, Gilbertson RJ, Schaiquevich P, Sun J et al. Pediatric phase I trial and pharmacokinetic study of vorinostat: a Children's Oncology Group phase I consortium report. J Clin Oncol 2010; 28: 3623–3629.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Hummel TR, Wagner L, Ahern C, Fouladi M, Reid JM, McGovern RM et al. A pediatric phase 1 trial of vorinostat and temozolomide in relapsed or refractory primary brain or spinal cord tumors: a Children's Oncology Group Phase 1 Consortium Study. Pediatr Blood Cancer 2013; 60: 1452–1457.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Su JM, Li XN, Thompson P, Ou CN, Ingle AM, Russell H et al. Phase 1 study of valproic acid in pediatric patients with refractory solid or CNS tumors: a children's oncology group report. Clin Cancer Res 2010; 17: 589–597.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Machida Y, Murai K, Miyake K, Iijima S . Expression of chromatin remodeling factors during neural differentiation. J Biochem 2001; 129: 43–49.

    Article  CAS  PubMed  Google Scholar 

  38. Keshelava N, Seeger RC, Groshen S, Reynolds CP . Drug resistance patterns of human neuroblastoma cell lines derived from patients at different phases of therapy. Cancer Res 1998; 58: 5396–5405.

    CAS  PubMed  Google Scholar 

  39. Liu X, Tian X, Wang F, Ma Y, Kornmann M, Yang Y . BRG1 promotes chemoresistance of pancreatic cancer cells through crosstalking with Akt signalling. Eur J Cancer 2014; 50: 2251–2262.

    Article  CAS  PubMed  Google Scholar 

  40. Opel D, Poremba C, Simon T, Debatin KM, Fulda S . Activation of Akt predicts poor outcome in neuroblastoma. Cancer Res 2007; 67: 735–745.

    Article  CAS  PubMed  Google Scholar 

  41. Chesler L, Schlieve C, Goldenberg DD, Kenney A, Kim G, McMillan A et al. Inhibition of phosphatidylinositol 3-kinase destabilizes Mycn protein and blocks malignant progression in neuroblastoma. Cancer Res 2006; 66: 8139–8146.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Bender A, Opel D, Naumann I, Kappler R, Friedman L, von Schweinitz D et al. PI3K inhibitors prime neuroblastoma cells for chemotherapy by shifting the balance towards pro-apoptotic Bcl-2 proteins and enhanced mitochondrial apoptosis. Oncogene 2011; 30: 494–503.

    Article  CAS  PubMed  Google Scholar 

  43. Opel D, Naumann I, Schneider M, Bertele D, Debatin KM, Fulda S . Targeting aberrant PI3K/Akt activation by PI103 restores sensitivity to TRAIL-induced apoptosis in neuroblastoma. Clin Cancer Res 2011; 17: 3233–3247.

    Article  CAS  PubMed  Google Scholar 

  44. Lamers F, Schild L, den Hartog IJ, Ebus ME, Westerhout EM, Ora I et al. Targeted BCL2 inhibition effectively inhibits neuroblastoma tumour growth. Eur J Cancer 2012; 48: 3093–3103.

    Article  CAS  PubMed  Google Scholar 

  45. Boix J, Fibla J, Yuste V, Piulats JM, Llecha N, Comella JX . Serum deprivation and protein synthesis inhibition induce two different apoptotic processes in N18 neuroblastoma cells. Exp Cell Res 1998; 238: 422–429.

    Article  CAS  PubMed  Google Scholar 

  46. Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263–267.

    Article  CAS  PubMed  Google Scholar 

  47. Zufferey R, Dull T, Mandel RJ, Bukovsky A, Quiroz D, Naldini L et al. Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 1998; 72: 9873–9880.

    CAS  PubMed  PubMed Central  Google Scholar 

  48. Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001; 25: 402–408.

    Article  CAS  PubMed  Google Scholar 

  49. Yuste VJ, Bayascas JR, Llecha N, Sanchez-Lopez I, Boix J, Comella JX . The absence of oligonucleosomal DNA fragmentation during apoptosis of IMR-5 neuroblastoma cells: disappearance of the caspase-activated DNase. J Biol Chem 2001; 276: 22323–22331.

    Article  CAS  PubMed  Google Scholar 

  50. Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 2003; 4: 249–264.

    Article  PubMed  Google Scholar 

  51. Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM et al. Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet 2000; 25: 25–29.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Kanehisa M, Goto S . KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res 2000; 28: 27–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Kanehisa M, Goto S, Sato Y, Furumichi M, Tanabe M . KEGG for integration and interpretation of large-scale molecular data sets. Nucleic Acids Res 2012; 40: D109–D114.

    Article  CAS  PubMed  Google Scholar 

  54. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005; 102: 15545–15550.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Saeed AI, Sharov V, White J, Li J, Liang W, Bhagabati N et al. TM4: a free, open-source system for microarray data management and analysis. Biotechniques 2003; 34: 374–378.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank members of the Genomics Facility, Statitstics and Bioinformatic Unit, Pathology Laboratory and the Animal Core Facilities of the Vall d’Hebron Research Institute. We thank Dr Mireia Duñac, Dr Jose R Bayascas and Dr Diego Arango for precious advice. We thank Ms Christine O’Hara for text correction. This work was supported by the Instituto de Salud Carlos III (CP11/00052, RD12/0036/0016, RD12/0036/0020, RD12/0036/0045, RD12/0036/0012) co-financed by the European Regional Development Fund (ERDF), Generalitat de Catalunya 2014-SGR-660 and Marie Curie Career Integration Grants.

Author information

Authors and Affiliations

  1. Laboratory of Translational Research in Child and Adolescent Cancer. Vall d’Hebron Research Institute (VHIR)-UAB, Barcelona, Spain

    L Jubierre, A Soriano, L París-Coderch, A Almazán-Moga, C Molist, J Roma, J Sánchez de Toledo, S Gallego & M F Segura

  2. Cell Signaling and Apoptosis Group, VHIR-UAB, Barcelona, Spain

    L Planells-Ferrer, R S Moubarak, M Sánchez-Céspedes & J X Comella

  3. Vall d’Hebron Institut of Oncology (VHIO), Stem Cell and Cancer Laboratory, Barcelona, Spain

    S P Tenbaum & H G Palmer

  4. Epigenetic and Cancer Biology Program-PEBC/Bellvitge Biomedical Research Institute-IDIBELL Barcelona, Barcelona, Spain

    O A Romero

  5. School of Medicine, University of Valencia, Valencia, Spain

    S Navarro & R Noguera

Authors
  1. L Jubierre
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A Soriano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L Planells-Ferrer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. L París-Coderch
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S P Tenbaum
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. O A Romero
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. R S Moubarak
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. A Almazán-Moga
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. C Molist
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J Roma
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. S Navarro
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. R Noguera
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. M Sánchez-Céspedes
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. J X Comella
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. H G Palmer
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. J Sánchez de Toledo
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. S Gallego
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. M F Segura
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M F Segura.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jubierre, L., Soriano, A., Planells-Ferrer, L. et al. BRG1/SMARCA4 is essential for neuroblastoma cell viability through modulation of cell death and survival pathways. Oncogene 35, 5179–5190 (2016). https://doi.org/10.1038/onc.2016.50

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2016.50

This article is cited by

Search

Advanced search

Quick links