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Chromosomal heterogeneity and instability characterize pediatric medulloblastoma cell lines and affect neoplastic phenotype

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Abstract

Chromosomal heterogeneity is a hallmark of most tumors and it can drive critical events as growth advantages, survival advantages, progression and karyotypic evolution. Medulloblastoma (MB) is the most common malignant central nervous system tumor in children. This work attempted to investigate chromosomal heterogeneity and instability profiles of two MB pediatric cell lines and their relationship with cell phenotype. We performed GTG-banding and cytokinesis-block micronucleus cytome assays, as well as morphological characterization, cell population doubling time, colony-forming efficiency, and chemo-sensitivity assays in two pediatric MB cell lines (UW402 and UW473). Both MB cells showed a high chromosomal heterogeneity. UW473 cells showed ~2 fold higher both clonal- and non-clonal chromosomal alterations than UW402 cells. Besides, UW473 showed two clonal-groups well-differentiated by ploidy level (<2n> and <4n>) and also presented a significantly higher number of chromosomal instability biomarkers. These results were associated with high morphological heterogeneity and survival advantages for UW473 and proliferation advantages for UW402 cells. Moreover, UW473 was significantly more sensitive to methotrexate, temozolomide and cisplatin while UW402 cells were more sensitive to doxorubicin. These data suggest that distinct different degrees of karyotypic heterogeneity and instability may affect neoplasic phenotype of MB cells. These findings bring new insights into cell and tumor biology.

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References

  • Albertson DG, Collins C, McCormick F, Gray JW (2003) Chromosome aberrations in solid tumors. Nat Genet 34:369–376

    Article  CAS  Google Scholar 

  • Aldosari N, Wiltshire RN, Dutra A, Schrock E, McLendon RE, Friedman HS, Bigner DD, Bigner SH (2002) Comprehensive molecular cytogenetic investigation of chromosomal abnormalities in human medulloblastoma cell lines and xenograft. Neuro Oncol 4:75–85

    CAS  Google Scholar 

  • Batra SK, McLendon RE, Koo JS, Castelino-Prabhu S, Fuchs HE, Krischer JP, Friedman HS, Bigner DD, Bigner SH (1995) Prognostic implications of chromosome 17p deletions in human medulloblastomas. J Neurooncol 24:39–45

    Article  CAS  Google Scholar 

  • Bayani J, Zielenska M, Marrano P, Kwan Y, Taylor MD, Jay V, Rutka JT, Squire JA (2000) Molecular cytogenetic analysis of medulloblastomas and supratentorial primitive neuroectodermal tumors by using conventional banding, comparative genomic hybridization, and spectral karyotyping. J Neurosurg 93:437–448

    Article  CAS  Google Scholar 

  • Bayani J, Selvarajah S, Maire G, Vukovic B, Al-Romaih K, Zielenska M, Squire JA (2007) Genomic mechanisms and measurement of structural and numerical instability in cancer cells. Semin Cancer Biol 17:5–18

    Article  CAS  Google Scholar 

  • Biegel JA (1999) Cytogenetics and molecular genetics of childhood brain tumors. Neuro-oncol 1:139–151

    CAS  Google Scholar 

  • Binda E, Visioli A, Reynolds B, Vescovi AL (2012) Heterogeneity of cancer-Initiating cells within glioblastoma. Front Biosci (Schol Ed) 4:1235–1248

    Article  Google Scholar 

  • Birkbak NJ, Eklund AC, Li Q, McClelland SE, Endesfelder D, Tan P, Tan IB, Richardson AL, Szallasi Z, Swanton C (2011) Paradoxical relationship between chromosomal instability and survival outcome in cancer. Cancer Res 71:3447–3452

    Article  CAS  Google Scholar 

  • Brassesco MS, Castro-Gamero AM, Valera ET, Neder L, Elias J Jr, Tone LG (2009) 3q27 aberrations in a childhood ovary teratoma with associated malignant germ cell component. Pediatr Blood Cancer 52:398–401

    Article  Google Scholar 

  • Cahill DP, Kinzler KW, Vogelstein B, Lengauer C (1999) Genetic instability and darwinian selection in tumours. Trends Cell Biol 9:M57–M60

    Article  CAS  Google Scholar 

  • Carter SL, Eklund AC, Kohane IS, Harris LN, Szallasi Z (2006) A signature of chromosomal instability inferred from gene expression profiles predicts clinical outcome in multiple human cancers. Nat Genet 38:1043–1048

    Article  CAS  Google Scholar 

  • Chandhok NS, Pellman D (2009) A little CIN may cost a lot: revisiting aneuploidy and cancer. Current Curr Opin Genet Dev 19:74–81

    Article  CAS  Google Scholar 

  • Cho YJ, Tsherniak A, Tamayo P, Santagata S, Ligon A, Greulich H, Berhoukim R, Amani V, Goumnerova L, Eberhart CG, Lau CC, Olson JM, Gilbertson RJ, Gajjar A, Delattre O, Kool M, Ligon K, Meyerson M, Mesirov JP, Pomeroy SL (2011) Integrative genomic analysis of medulloblastoma identifies a molecular subgroup that drives poor clinical outcome. J Clin Oncol 29:1424–1430

    Article  Google Scholar 

  • Cohen N, Betts DR, Tavori U, Toren A, Ram T, Constantini S, Grotzer MA, Amariglio N, Rechavi G, Trakhtenbrot L (2004) Karyotypic evolution pathways in medulloblastoma/primitive neuroectodermal tumor determined with a combination of spectral karyotyping, G-banding, and fluorescence in situ hybridization. Cancer Genet Cytogenet 149:44–52

    Article  Google Scholar 

  • Eigen M (2002) Error catastrophe and antiviral strategy. Proc Natl Acad Sci USA 99:13374–13376

    Article  CAS  Google Scholar 

  • Fenech M (2007) Cytokinesis-block micronucleus cytome assay. Nat Protoc 2:1084–1104

    Article  CAS  Google Scholar 

  • Finlay JL, Erdreich-Epstein A, Packer RJ (2007) Progress in the treatment of childhood brain tumors: no room for complacency. Pediatr Hematol Oncol 24:79–84

    Article  Google Scholar 

  • Fischer SJ, Benson LM, Fauq A, Naylor S, Windebank AJ (2008) Cisplatin and dimethyl sulfoxide react to form an adducted compound with reduced cytotoxicity and neurotoxicity. Neurotoxicology 29:444–452

    Article  CAS  Google Scholar 

  • Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C (2006) Clonogenic assay of cells in vitro. Nat Protoc 1:2315–2319

    Article  CAS  Google Scholar 

  • Gilbertson RJ, Ellison DW (2008) The origins of medulloblastoma subtypes. Annu Rev Pathol 3:341–365

    Article  CAS  Google Scholar 

  • Gisselsson D (2008) Classification of chromosome segregation errors in cancer. Chromosoma 117:511–519

    Article  Google Scholar 

  • Gisselsson D, Pettersson L, Hoglund M, Heidenblad M, Gorunova L, Wiegant J, Mertens F, Dal Cin P, Mitelman F, Mandahl N (2000) Chromosomal breakage-fusion- bridge events cause genetic intratumor heterogeneity. Proc Natl Acad Sci USA 97:5357–5362

    Article  CAS  Google Scholar 

  • Glanz C, Rebetz J, Stewénius Y, Persson A, Englund E, Mandahl N, Mertens F, Salford LG, Widegren B, Fan X, Gisselsson D (2007) Genetic intratumour heterogeneity in high-grade brain tumours is associated with telomere-dependent mitotic instability. Neuropathol Appl Neurobiol 33:440–454

    Article  CAS  Google Scholar 

  • Gordon DJ, Resio B, Pellman D (2012) Causes and consequences of aneuploidy in cancer. Nat Rev Genet 13:189–203

    CAS  Google Scholar 

  • Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674

    Article  CAS  Google Scholar 

  • Heim S, Mitelman F (1994) Cancer cytogenetics. Alan R. Liss, Inc., New York, pp 23–37

    Google Scholar 

  • Heng HH, Bremer SW, Stevens J, Ye KJ, Miller F, Liu G, Ye CJ (2006a) Cancer progression by non-clonal chromosome aberrations. J Cell Biochem 98:1424–1435

    Article  CAS  Google Scholar 

  • Heng HH, Liu G, Bremer SW, Ye KJ, Stevens J, Ye CJ (2006b) Clonal and nonclonal chromosome aberrations and genome variation/aberration. Genome 49:195–204

    Article  CAS  Google Scholar 

  • Heng HH, Stevens JB, Liu G, Bremer SW, Ye KJ, Reddy PV, Wu GS, Wang YA, Tainsky MA, Ye CJ (2006c) Stochastic cancer progression driven by non-clonal chromosome aberrations. J Cell Physiol 208:461–472

  • Heng HH, Stevens J, Bremer SW, Ye KJ, Liu G, Ye CJ (2010) The evolutionary mechanism of cancer. J Cell Biochem 109:1072–1084

    CAS  Google Scholar 

  • Huse JT, Holland EC (2010) Targeting brain cancer: advances in the molecular pathology of malignant glioma and medulloblastoma. Nat Rev Cancer 10:319–331

    Article  CAS  Google Scholar 

  • Janssen A, Kops GJ, Medema RH (2009) Elevating the frequency of chromosome mis-segregation as a strategy to kill tumor cells. Proc Natl Acad Sci USA 106:19108–19113

    Article  CAS  Google Scholar 

  • Jones DT, Jäger N, Kool M, Zichner T, Hutter B, Sultan M, Cho YJ, Pugh TJ, Hovestadt V, Stütz AM, Rausch T, Warnatz HJ, Ryzhova M, Bender S, Sturm D, Pleier S, Cin H, Pfaff E, Sieber L, Wittmann A, Remke M, Witt H, Hutter S, Tzaridis T, Weischenfeldt J, Raeder B, Avci M, Amstislavskiy V, Zapatka M, Weber UD, Wang Q, Lasitschka B, Bartholomae CC, Schmidt M, von Kalle C, Ast V, Lawerenz C, Eils J, Kabbe R, Benes V, van Sluis P, Koster J, Volckmann R, Shih D, Betts MJ, Russell RB, Coco S, Tonini GP, Schüller U, Hans V, Graf N, Kim YJ, Monoranu C, Roggendorf W, Unterberg A, Herold-Mende C, Milde T, Kulozik AE, von Deimling A, Witt O, Maass E, Rössler J, Ebinger M, Schuhmann MU, Frühwald MC, Hasselblatt M, Jabado N, Rutkowski S, von Bueren AO, Williamson D, Clifford SC, McCabe MG, Collins VP, Wolf S, Wiemann S, Lehrach H, Brors B, Scheurlen W, Felsberg J, Reifenberger G, Northcott PA, Taylor MD, Meyerson M, Pomeroy SL, Yaspo ML, Korbel JO, Korshunov A, Eils R, Pfister SM, Lichter P (2012) Dissecting the genomic complexity underlying medulloblastoma. Nature 488:100–105

    Article  CAS  Google Scholar 

  • Keles GE, Berger MS, Srinivasan J, Kolstoe DD, Bobola MS, Silber JR (1995) Establishment and characterization of four human medulloblastoma-derived cell lines. Oncol Res 7:493–503

    CAS  Google Scholar 

  • Könemann S, Schuck A, Malath J et al (2000) Cell heterogeneity and subpopulations in solid tumors characterized by simultaneous immunophenotyping and DNA content analysis. Cytometry 41:172–177

    Article  Google Scholar 

  • Kool M, Koster J, Bunt J, Hasselt NE, Lakeman A, van Sluis P, Troost D, Meeteren NS, Caron HN, Cloos J, Mrsic A, Ylstra B, Grajkowska W, Hartmann W, Pietsch T, Ellison D, Clifford SC, Versteeg R (2008) Integrated genomics identifies five medulloblastoma subtypes with distinct genetic profiles, pathway signatures and clinicopathological features. PLoS ONE 3:e3088

    Article  Google Scholar 

  • Kool M, Korshunov A, Remke M, Jones D, Schlanstein M, Northcott P, Cho Y, Schouten-van Meeteren N, van Vuurden D, Clifford S, Pietsch T, von Bueren A, Rutkowski S, McCabe M, Collins P, Bäcklund M, Haberler C, Bourdeaut F, Delattre O, Doz F, Ellison D, Gilbertson R, Pomeroy S, Taylor M, Lichter P, Pfister S (2012) Molecular subgroups of medulloblastoma: an international meta-analysis of transcriptome, genetic aberrations, and clinical data of wnt, shh, group 3, and group 4 medulloblastomas. Acta Neuropathol 123:473–484

    Article  CAS  Google Scholar 

  • Kops GJ, Foltz DR, Cleveland DW (2004) Lethality to human cancer cells through massive chromosome loss by inhibition of the mitotic checkpoint. Proc Natl Acad Sci USA 101:8699–8704

    Article  CAS  Google Scholar 

  • Lamont JM, McManamy CS, Pearson AD, Clifford SC, Ellison DW (2004) Combined histopathological and molecular cytogenetic stratification of medulloblastoma patients. Clin Cancer Res 10:5482–5493

    Article  CAS  Google Scholar 

  • Lee AJX, Swanton C (2012) Tumour heterogeneity and drug resistance: personalising cancer medicine through functional genomics. Biochem Pharmacol 83:1013–1020

    Article  CAS  Google Scholar 

  • Lee AJX, Endesfelder D, Rowan AJ, Walther A, Birkbak NJ, Futreal PA, Downward J, Szallasi Z, Tomlinson IPM, Kschischo M, Swanton C (2011) Chromosomal Instability Confers Intrinsic Multi-Drug Resistance. Cancer Res 71:1858–1870

    Article  CAS  Google Scholar 

  • Lo KC, Ma C, Bundy BN, Pomeroy SL, Eberhart CG, Cowell JK (2007) Gain of 1q is a potential univariate negative prognostic marker for survival in medulloblastoma. Clin Cancer Res 13:7022–7028

    Article  CAS  Google Scholar 

  • Louis DN, Ohgaki H, Wiestler OD, Cavenee WK (2007) WHO classification of tumors of the central nervous system. International Agency for Research on Cancer (IARC), Lyon

  • Lynch M, Burger R, Butcher D, Gabriel W (1993) The mutational meltdown in asexual populations. J Hered 84:339–344

    CAS  Google Scholar 

  • McCabe MG, Bäcklund LM, Leong HS, Ichimura K, Collins VP (2011) Chromosome 17 alterations identify good-risk and poor-risk tumors independently of clinical factors in medulloblastoma. Neuro Oncol 13:376–383

    Article  CAS  Google Scholar 

  • McGranahan N, Burrell RA, Endesfelder D, Novelli MR, Swanton C (2012) Cancer chromosomal instability: therapeutic and diagnostic challenges. EMBO Rep 13:528–538

    Article  CAS  Google Scholar 

  • Mitelman F (2000) Recurrent chromosome aberrations in cancer. Mutat Res 462:247–253

    Article  CAS  Google Scholar 

  • Nicholson JM, Duesberg P (2009) On the karyotypic origin and evolution of cancer cells. Cancer Genet Cytogenet 194:96–110

    Article  CAS  Google Scholar 

  • Northcott PA, Korshunov A, Witt H, Hielscher T, Eberhart CG, Mack S, Bouffet E, Clifford SC, Hawkins CE, French P, Rutka JT, Pfister S, Taylor MD (2011) Medulloblastoma comprises four distinct molecular variants. J Clin Oncol 29:1408–1414

    Article  Google Scholar 

  • Northcott PA, Jones DT, Kool M, Robinson GW, Gilbertson RJ, Cho YJ, Pomeroy SL, Korshunov A, Lichter P, Taylor MD, Pfister SM (2012) Medulloblastomics: the end of the beginning. Nat Rev Cancer 12:818–834

    Article  CAS  Google Scholar 

  • Packer R, Vezina G (2008) Management of and prognosis with medulloblastoma: therapy at a crossroads. Arch Neurol 65:1419–1424

    Article  Google Scholar 

  • Pan E, Pellarin M, Holmes E, Smirnov I, Misra A, Eberhart CG, Burger PC, Biegel JA, Feuerstein BG (2005) Isochromosome 17q is a negative prognostic factor in poor-risk childhood medulloblastoma patients. Clin Cancer Res 11:4733–4740

    Article  CAS  Google Scholar 

  • Pfister S, Remke M, Benner A, Mendrzyk F, Toedt G, Felsberg J, Wittmann A, Devens F, Gerber NU, Joos S, Kulozik A, Reifenberger G, Rutkowski S, Wiestler OD, Radlwimmer B, Scheurlen W, Lichter P, Korshunov A (2009) Outcome prediction in pediatric medulloblastoma based on DNA copy-number aberrations of chromosomes 6q and 17q and the MYC and MYCN loci. J Clin Oncol 27:1627–1636

    Article  Google Scholar 

  • Pomeroy SL, Tamayo P, Gaasenbeek M, Sturla LM, Angelo M, McLaughlin ME, Kim JY, Goumnerova LC, Black PM, Lau C, Allen JC, Zagzag D, Olson JM, Curran T, Wetmore C, Biegel JA, Poggio T, Mukherjee S, Rifkin R, Califano A, Stolovitzky G, Louis DN, Mesirov JP, Lander ES, Golub TR (2002) Prediction of central nervous system embryonal tumour outcome based on gene expression. Nature 415:436–442

    Article  CAS  Google Scholar 

  • Read TA, Hegedus B, Wechsler-Reya R, Gutmann DH (2006) The neurobiology of neurooncology. Ann Neurol 60:3–11

    Article  CAS  Google Scholar 

  • Remke M, Hielscher T, Korshunov A, Northcott PA, Bender S, Kool M, Westermann F, Benner A, Cin H, Ryzhova M, Sturm D, Witt H, Haag D, Toedt G, Wittmann A, Schottler A, von Bueren AO, von Deimling A, Rutkowski S, Scheurlen W, Kulozik AE, Taylor MD, Lichter P, Pfister SM (2011a) FSTL5 is a marker of poor prognosis in non-WNT/Non-SHH medulloblastoma. J Clin Oncol 29:3852–3861

    Article  CAS  Google Scholar 

  • Remke M, Hielscher T, Northcott PA, Witt H, Ryzhova M, Wittmann A, Benner A, von Deimling A, Scheurlen W, Perry A, Croul S, Kulozik AE, Lichter P, Taylor MD, Pfister SM, Korshunov A (2011b) Adult medulloblastoma comprises three major molecular variants. J Clin Oncol 29:2717–2723

    Article  Google Scholar 

  • Roschke AV, Tonon G, Gehlhaus KS, McTyre N, Bussey KJ, Lababidi S, Scudiero DA, Weinstein JN, Kirsch IR (2003) Karyotypic complexity of the NCI-60 drug-screening panel. Cancer Res 63:8634–8647

    CAS  Google Scholar 

  • Roylance R, Endesfelder D, Gorman P, Burrell RA, Sander J, Tomlinson I, Hanby AM, Speirs V, Richardson AL, Birkbak NJ, Eklund AC, Downward J, Kschischo M, Szallasi Z, Swanton C (2011) Relationship of extreme chromosomal instability with longterm survival in a retrospective analysis of primary breast cancer. Cancer Epidemiol Biomarkers Prev 20:2183–2194

    Article  Google Scholar 

  • Selvarajah S, Yoshimoto M, Park PC, Maire G, Paderova J, Bayani J, Lim G, Al-Romaih K, Squire JA, Zielenska M (2006) The breakage-fusionbridge (BFB) cycle as a mechanism for generating genetic heterogeneity in osteosarcoma. Chromosoma 115:459–467

    Article  CAS  Google Scholar 

  • Shaffer LG, Tommerup N (2005) An international system for human cytogenetic nomenclature (2005). Karger Publishers, Basel

    Google Scholar 

  • Sheltzer JM, Amon A (2011) The aneuploidy paradox: costs and benefits of an incorrect karyotype. Trends Genet 27:446–453

    Article  CAS  Google Scholar 

  • Shimizu N, Shingaki K, Kaneko-Sasaguri Y, Hashizume T, Kanda T (2005) When, where and how the bridge breaks: anaphase bridge breakage plays a crucial role in gene amplification and HSR generation. Exp Cell Res 302:233–243

    Article  CAS  Google Scholar 

  • Taylor MD, Northcott PA, Korshunov A, Remke M, Cho YJ, Clifford SC, Eberhart CG, Parsons W, Rutkowski S, Gajjar A, Ellison DW, Lichter P, Gilbertson RJ, Pomeroy SL, Pfister SL, Kool M (2012) Molecular subgroups of medulloblastoma: the current consensus. Acta Neuropathol 123:465–472

    Article  CAS  Google Scholar 

  • Thompson MC, Fuller C, Hogg TL, Dalton J, Finkelstein D, Lau CC, Chintagumpala M, Adesina A, Ashley DM, Kellie SJ, Taylor MD, Curran T, Gajjar A, Gilbertson RJ (2006) Genomics identifies medulloblastoma subgroups that are enriched for specific genetic alterations. J Clin Oncol 24:1924–1931

    Article  CAS  Google Scholar 

  • Torres EM, Sokolsky T, Tucker CM, Chan LY, Boselli M, Dunham MJ, Amon A (2007) Effects of aneuploidy on cellular physiology and cell division in haploid yeast. Science 317:916–924

    Article  CAS  Google Scholar 

  • Williams BR, Prabhu VR, Hunter KE, Glazier CM, Whittaker CA, Housman DE, Amon A (2008) Aneuploidy affects proliferation and spontaneous immortalization in mammalian cells. Science 322:703–709

    Article  CAS  Google Scholar 

  • Ye CJ, Liu G, Bremer SW, Heng HH (2007) The dynamics of cancer chromosomes and genomes. Cytogenet Genome Res 118:237–246

    Article  CAS  Google Scholar 

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Acknowledgments

The authors thank Patrícia Vianna Bonini Palma, Camila Cristina de Oliveira Menezes Bonaldo, and Daiane Fernanda dos Santos, Hemocentro-FMRP-USP, Ribeirão Preto, Brazil, Brazil, for assistance with the flow cytometry. The authors also thank Coordenação para o Aperfeiçoamento de Pessoal em Nível Superior-Brazil (CAPES) and Fundação de Apoio ao Ensino, Pesquisa e Assistência do Hospital das Clínicas da FMRPUSP (FAEPA) for financial support.

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The authors declare that they have no conflict of interest.

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Correspondence to Angel Mauricio Castro-Gamero.

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Castro-Gamero, A.M., Borges, K.S., Lira, R.C. et al. Chromosomal heterogeneity and instability characterize pediatric medulloblastoma cell lines and affect neoplastic phenotype. Cytotechnology 65, 871–885 (2013). https://doi.org/10.1007/s10616-012-9529-z

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