Skip to main content

Advertisement

SpringerLink
Log in

Alternative lengthening of telomeres in neuroblastoma cell lines is associated with a lack of MYCN genomic amplification and with p53 pathway aberrations

  • Laboratory Investigation
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Alternative lengthening of telomeres (ALT) is a telomerase-independent telomere length maintenance mechanism that enables the unlimited proliferation of a subset of cancer cells. Some neuroblastoma (NB) tumors appear to maintain telomere length by activating ALT. Of 40 NB cell lines, we identified four potential ALT cell lines (CHLA-90, SK-N-FI, LA-N-6, and COG-N-291) that were telomerase-negative and had long telomeres (a feature of ALT cells). All four cell lines lacked MYCN amplification and were p53 non-functional upon irradiation. Two of these cell lines (CHLA-90 and SK-N-FI) were positive for C-circles (telomeric DNA circles) and ALT-associated promyelocytic leukemia nuclear bodies, both of which are phenotypic characteristics of ALT. Mutation of ATRX (associated with ALT in tumors) was only found in CHLA-90. Thus, the ALT phenotype in NB may not be limited to tumors with ATRX mutations but is associated with a lack of MYCN amplification and alterations in the p53 pathway.

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

Similar content being viewed by others

References

  1. Blackburn EH (1991) Structure and function of telomeres. Nature 350:569–573

    Article  CAS  PubMed  Google Scholar 

  2. O’Sullivan RJ, Karlseder J (2010) Telomeres: protecting chromosomes against genome instability. Nat Rev Mol Cell Biol 11:171–181

    PubMed Central  PubMed  Google Scholar 

  3. Harley CB, Futcher AB, Greider CW (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460

    Article  CAS  PubMed  Google Scholar 

  4. d’Adda di Fagagna F, Reaper PM, Clay-Farrace L, Fiegler H, Carr P, Von Zglinicki T, Saretzki G, Carter NP, Jackson SP (2003) A DNA damage checkpoint response in telomere-initiated senescence. Nature 426:194–198

    Article  PubMed  Google Scholar 

  5. Greider CW (1998) Telomerase activity, cell proliferation, and cancer. Proc Natl Acad Sci USA 95:90–92

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Greider CW, Blackburn EH (1985) Identification of a specific telomere terminal transferase activity in tetrahymena extracts. Cell 43:405–413

    Article  CAS  PubMed  Google Scholar 

  7. Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL, Shay JW (1994) Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015

    Article  CAS  PubMed  Google Scholar 

  8. Bryan TM, Englezou A, Dalla-Pozza L, Dunham MA, Reddel RR (1997) Evidence for an alternative mechanism for maintaining telomere length in human tumors and tumor-derived cell lines. Nat Med 3:1271–1274

    Article  CAS  PubMed  Google Scholar 

  9. Bryan TM, Englezou A, Gupta J, Bacchetti S, Reddel RR (1995) Telomere elongation in immortal human cells without detectable telomerase activity. EMBO J 14:4240–4248

    CAS  PubMed Central  PubMed  Google Scholar 

  10. Cesare AJ, Reddel RR (2010) Alternative lengthening of telomeres: models, mechanisms and implications. Nat Rev Genet 11:319–330

    Article  CAS  PubMed  Google Scholar 

  11. Henson JD, Reddel RR (2010) Assaying and investigating alternative lengthening of telomeres activity in human cells and cancers. FEBS Lett 584:3800–3811

    Article  CAS  PubMed  Google Scholar 

  12. Henson JD, Neumann AA, Yeager TR, Reddel RR (2002) Alternative lengthening of telomeres in mammalian cells. Oncogene 21:598–610

    Article  CAS  PubMed  Google Scholar 

  13. Yeager TR, Neumann AA, Englezou A, Huschtscha LI, Noble JR, Reddel RR (1999) Telomerase-negative immortalized human cells contain a novel type of promyelocytic leukemia (PML) body. Cancer Res 59:4175–4179

    CAS  PubMed  Google Scholar 

  14. Nabetani A, Ishikawa F (2011) Alternative lengthening of telomeres pathway: recombination-mediated telomere maintenance mechanism in human cells. J Biochem 149:5–14

    Article  CAS  PubMed  Google Scholar 

  15. Henson JD, Cao Y, Huschtscha LI, Chang AC, Au AY, Pickett HA, Reddel RR (2009) DNA C-circles are specific and quantifiable markers of alternative-lengthening-of-telomeres activity. Nat Biotechnol 27:1181–1185

    Article  CAS  PubMed  Google Scholar 

  16. Heaphy CM, de Wilde RF, Jiao Y, Klein AP, Edil BH, Shi C, Bettegowda C, Rodriguez FJ, Eberhart CG, Hebbar S, Offerhaus GJ, McLendon R, Rasheed BA, He Y, Yan H, Bigner DD, Oba-Shinjo SM, Marie SK, Riggins GJ, Kinzler KW, Vogelstein B, Hruban RH, Maitra A, Papadopoulos N, Meeker AK (2011) Altered telomeres in tumors with ATRX and DAXX mutations. Science 333:425

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Wong LH, McGhie JD, Sim M, Anderson MA, Ahn S, Hannan RD, George AJ, Morgan KA, Mann JR, Choo KH (2010) ATRX interacts with H3.3 in maintaining telomere structural integrity in pluripotent embryonic stem cells. Genome Res 20:351–360

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Lewis PW, Elsaesser SJ, Noh KM, Stadler SC, Allis CD (2010) Daxx is an H3.3-specific histone chaperone and cooperates with ATRX in replication-independent chromatin assembly at telomeres. Proc Natl Acad Sci USA 107:14075–14080

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  19. Cheung NK, Zhang J, Lu C, Parker M, Bahrami A, Tickoo SK, Heguy A, Pappo AS, Federico S, Dalton J, Cheung IY, Ding L, Fulton R, Wang J, Chen X, Becksfort J, Wu J, Billups CA, Ellison D, Mardis ER, Wilson RK, Downing JR, Dyer MA (2012) Association of age at diagnosis and genetic mutations in patients with neuroblastoma. JAMA 307:1062–1071

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  20. Molenaar JJ, Koster J, Zwijnenburg DA, van Sluis P, Valentijn LJ, van der Ploeg I, Hamdi M, van Nes J, Westerman BA, van Arkel J, Ebus ME, Haneveld F, Lakeman A, Schild L, Molenaar P, Stroeken P, van Noesel MM, Ora I, Santo EE, Caron HN, Westerhout EM, Versteeg R (2012) Sequencing of neuroblastoma identifies chromothripsis and defects in neuritogenesis genes. Nature 483:589–593

    Article  CAS  PubMed  Google Scholar 

  21. Pugh TJ, Morozova O, Attiyeh EF, Asgharzadeh S, Wei JS, Auclair D, Carter SL, Cibulskis K, Hanna M, Kiezun A, Kim J, Lawrence MS, Lichenstein L, McKenna A, Pedamallu CS, Ramos AH, Shefler E, Sivachenko A, Sougnez C, Stewart C, Ally A, Birol I, Chiu R, Corbett RD, Hirst M, Jackman SD, Kamoh B, Khodabakshi AH, Krzywinski M, Lo A, Moore RA, Mungall KL, Qian J, Tam A, Thiessen N, Zhao Y, Cole KA, Diamond M, Diskin SJ, Mosse YP, Wood AC, Ji L, Sposto R, Badgett T, London WB, Moyer Y, Gastier-Foster JM, Smith MA, Auvil JM, Gerhard DS, Hogarty MD, Jones SJ, Lander ES, Gabriel SB, Getz G, Seeger RC, Khan J, Marra MA, Meyerson M, Maris JM (2013) The genetic landscape of high-risk neuroblastoma. Nat Genet 45:279–284

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  23. Onitake Y, Hiyama E, Kamei N, Yamaoka H, Sueda T, Hiyama K (2009) Telomere biology in neuroblastoma: telomere binding proteins and alternative strengthening of telomeres. J Pediatr Surg 44:2258–2266

    Article  PubMed  Google Scholar 

  24. Lundberg G, Sehic D, Lansberg JK, Ora I, Frigyesi A, Castel V, Navarro S, Piqueras M, Martinsson T, Noguera R, Gisselsson D (2011) Alternative lengthening of telomeres-An enhanced chromosomal instability in aggressive non-MYCN amplified and telomere elongated neuroblastomas. Genes Chromosomes Cancer. doi:10.1002/gcc.20850

    PubMed  Google Scholar 

  25. Ambros PF, Ambros IM, Brodeur GM, Haber M, Khan J, Nakagawara A, Schleiermacher G, Speleman F, Spitz R, London WB, Cohn SL, Pearson AD, Maris JM (2009) International consensus for neuroblastoma molecular diagnostics: report from the International Neuroblastoma Risk Group (INRG) Biology Committee. Br J Cancer 100:1471–1482

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Reynolds CP, Tomayko MM, Donner L, Helson L, Seeger RC, Triche TJ, Brodeur GM (1988) Biological classification of cell lines derived from human extra-cranial neural tumors. Prog Clin Biol Res 271:291–306

    CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  28. Keshelava N, Zuo JJ, Chen P, Waidyaratne SN, Luna MC, Gomer CJ, Triche TJ, Reynolds CP (2001) Loss of p53 function confers high-level multidrug resistance in neuroblastoma cell lines. Cancer Res 61:6185–6193

    CAS  PubMed  Google Scholar 

  29. Masters JR, Thomson JA, Daly-Burns B, Reid YA, Dirks WG, Packer P, Toji LH, Ohno T, Tanabe H, Arlett CF, Kelland LR, Harrison M, Virmani A, Ward TH, Ayres KL, Debenham PG (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc Natl Acad Sci USA 98:8012–8017

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  30. Lau LM, Dagg RA, Henson JD, Au AY, Royds JA, Reddel RR (2013) Detection of alternative lengthening of telomeres by telomere quantitative PCR. Nucleic Acids Res 41:e34

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  31. Mosse YP, Diskin SJ, Wasserman N, Rinaldi K, Attiyeh EF, Cole K, Jagannathan J, Bhambhani K, Winter C, Maris JM (2007) Neuroblastomas have distinct genomic DNA profiles that predict clinical phenotype and regional gene expression. Genes Chromosomes Cancer 46:936–949

    Article  CAS  PubMed  Google Scholar 

  32. Perrem K, Colgin LM, Neumann AA, Yeager TR, Reddel RR (2001) Coexistence of alternative lengthening of telomeres and telomerase in hTERT-transfected GM847 cells. Mol Cell Biol 21:3862–3875

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  33. Keshelava N, Tsao-Wei D, Reynolds CP (2003) Pyrazoloacridine is active in multidrug-resistant neuroblastoma cell lines with nonfunctional p53. Clin Cancer Res 9:3492–3502

    CAS  PubMed  Google Scholar 

  34. Jiao Y, Shi C, Edil BH, de Wilde RF, Klimstra DS, Maitra A, Schulick RD, Tang LH, Wolfgang CL, Choti MA, Velculescu VE, Diaz LA Jr, Vogelstein B, Kinzler KW, Hruban RH, Papadopoulos N (2011) DAXX/ATRX, MEN1, and mTOR pathway genes are frequently altered in pancreatic neuroendocrine tumors. Science 331:1199–1203

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Cawthon RM (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res 30:e47

    Article  PubMed Central  PubMed  Google Scholar 

  36. Binz N, Shalaby T, Rivera P, Shin-ya K, Grotzer MA (2005) Telomerase inhibition, telomere shortening, cell growth suppression and induction of apoptosis by telomestatin in childhood neuroblastoma cells. Eur J Cancer 41:2873–2881

    Article  CAS  PubMed  Google Scholar 

  37. Schmidt ML, Lukens JN, Seeger RC, Brodeur GM, Shimada H, Gerbing RB, Stram DO, Perez C, Haase GM, Matthay KK (2000) Biologic factors determine prognosis in infants with stage IV neuroblastoma: a prospective Children’s Cancer Group study. J Clin Oncol 18:1260–1268

    CAS  PubMed  Google Scholar 

  38. Lovejoy CA, Li W, Reisenweber S, Thongthip S, Bruno J, de Lange T, De S, Petrini JH, Sung PA, Jasin M, Rosenbluh J, Zwang Y, Weir BA, Hatton C, Ivanova E, Macconaill L, Hanna M, Hahn WC, Lue NF, Reddel RR, Jiao Y, Kinzler K, Vogelstein B, Papadopoulos N, Meeker AK (2012) Loss of ATRX, genome instability, and an altered DNA damage response are hallmarks of the alternative lengthening of telomeres pathway. PLoS Genet 8:e1002772

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  39. Carr J, Bell E, Pearson AD, Kees UR, Beris H, Lunec J, Tweddle DA (2006) Increased frequency of aberrations in the p53/MDM2/p14(ARF) pathway in neuroblastoma cell lines established at relapse. Cancer Res 66:2138–2145

    Article  CAS  PubMed  Google Scholar 

  40. Carr-Wilkinson J, O’Toole K, Wood KM, Challen CC, Baker AG, Board JR, Evans L, Cole M, Cheung NK, Boos J, Kohler G, Leuschner I, Pearson AD, Lunec J, Tweddle DA (2010) High frequency of p53/MDM2/p14ARF pathway abnormalities in relapsed neuroblastoma. Clin Cancer Res 16:1108–1118

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  41. Van Maerken T, Vandesompele J, Rihani A, De Paepe A, Speleman F (2009) Escape from p53-mediated tumor surveillance in neuroblastoma: switching off the p14(ARF)-MDM2-p53 axis. Cell Death Differ 16:1563–1572

    Article  PubMed  Google Scholar 

  42. Thompson PM, Maris JM, Hogarty MD, Seeger RC, Reynolds CP, Brodeur GM, White PS (2001) Homozygous deletion of CDKN2A (p16INK4a/p14ARF) but not within 1p36 or at other tumor suppressor loci in neuroblastoma. Cancer Res 61:679–686

    CAS  PubMed  Google Scholar 

  43. Salvioli S, Bonafe M, Barbi C, Storci G, Trapassi C, Tocco F, Gravina S, Rossi M, Tiberi L, Mondello C, Monti D, Franceschi C (2005) p53 codon 72 alleles influence the response to anticancer drugs in cells from aged people by regulating the cell cycle inhibitor p21WAF1. Cell Cycle 4:1264–1271

    Article  CAS  PubMed  Google Scholar 

  44. Hiyama E, Hiyama K, Yokoyama T, Matsuura Y, Piatyszek MA, Shay JW (1995) Correlating telomerase activity levels with human neuroblastoma outcomes. Nat Med 1:249–255

    Article  CAS  PubMed  Google Scholar 

  45. Poremba C, Willenbring H, Hero B, Christiansen H, Schafer KL, Brinkschmidt C, Jurgens H, Bocker W, Dockhorn-Dworniczak B (1999) Telomerase activity distinguishes between neuroblastomas with good and poor prognosis. Ann Oncol 10:715–721

    Article  CAS  PubMed  Google Scholar 

  46. Poremba C, Scheel C, Hero B, Christiansen H, Schaefer KL, Nakayama J, Berthold F, Juergens H, Boecker W, Dockhorn-Dworniczak B (2000) Telomerase activity and telomerase subunits gene expression patterns in neuroblastoma: a molecular and immunohistochemical study establishing prognostic tools for fresh-frozen and paraffin-embedded tissues. J Clin Oncol 18:2582–2592

    CAS  PubMed  Google Scholar 

  47. Bryan TM, Marusic L, Bacchetti S, Namba M, Reddel RR (1997) The telomere lengthening mechanism in telomerase-negative immortal human cells does not involve the telomerase RNA subunit. Hum Mol Genet 6:921–926

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a Grant from the National Cancer Institute (CA82830; C. P. Reynolds), Cancer Prevention & Research Institute of Texas (RP 110763; C. P. Reynolds), and Cure Cancer Australia Foundation (L. Lau); by a Cancer Institute of New South Wales Clinical Research Fellowship, Australia (07/CRF/105; L Lau) and National Health and Medical Research Council Early Career Fellowship, Australia (APP1012500; L. Lau).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Cancer Center, Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, TX, USA

    Ahsan S. Farooqi & C. Patrick Reynolds

  2. Department of Pharmacology, Texas Tech University Health Sciences Center, Lubbock, TX, USA

    C. Patrick Reynolds

  3. Department of Internal Medicine, Texas Tech University Health Sciences Center, Lubbock, TX, USA

    C. Patrick Reynolds

  4. Department of Pediatrics, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX, USA

    C. Patrick Reynolds

  5. Children’s Cancer Research Unit, Kids Research Institute, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia

    Rebecca A. Dagg & Loretta M. S. Lau

  6. Spectrum Pharmaceuticals, Inc., Irvine, CA, USA

    L. Mi Rim Choi

  7. Department of Cell Biology, Southwestern Medical Center, University of Texas, Dallas, TX, USA

    Jerry W. Shay

  8. Center of Excellence in Genomic Medicine Research, King Abdulaziz University, Jeddah, Saudi Arabia

    Jerry W. Shay

  9. Discipline of Paediatrics and Child Health, Sydney Medical School, University of Sydney, Sydney, NSW, Australia

    Loretta M. S. Lau

Authors
  1. Ahsan S. Farooqi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rebecca A. Dagg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L. Mi Rim Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jerry W. Shay
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C. Patrick Reynolds
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Loretta M. S. Lau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Loretta M. S. Lau.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Farooqi, A.S., Dagg, R.A., Choi, L.M.R. et al. Alternative lengthening of telomeres in neuroblastoma cell lines is associated with a lack of MYCN genomic amplification and with p53 pathway aberrations. J Neurooncol 119, 17–26 (2014). https://doi.org/10.1007/s11060-014-1456-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-014-1456-8

Keywords

Search

Navigation