Skip to main content
SpringerLink
Log in

In vitro properties of a newly established medulloblastoma cell line, MCD-1

  • Published:
Molecular and Chemical Neuropathology

Abstract

Medulloblastomas are poorly differentiated brain tumors believed to arise from primitive pleuripotential stem cells, and tend to express mixed neuronal and glial properties. In the present study, we examined immunohistochemical and neurotransmitter phenotypic properties in a newly established medulloblastoma cell line, MCD-1. MCD-1 cells were immortal, not contact-inhibited, but did not grow in soft agar. Immunohistochemical studies showed positive staining for neurofilament protein (NF), neuron-specific enolase (NSE), synaptophysin, MAP 2, τ, NCAM 180, vimentin, and S-100 protein. The cells expressed specific uptake of glutamate, serotonin, and choline, but not GABA or dopamine. A significant increase in process extension was seen in response to agents that enhance intracellular cyclic AMP, especially 3-isobutyl-1-methylxanthine (IBMX). Process formation induced by IBMX was associated with a decrease in cell proliferation as evidenced by a reduction in numbers of cells incorporating 5-bromo-2-deoxyuridine (BrdU). No increase in process extension was observed following exposure to NGF or retinoic acid. MCD-1 cells were shown to produce transforming growth factor beta (TGFβ), and were immunopositive for mutant p53. Transfection assays with the PG13-Luc reporter plasmid, which contains a p53-responsive enhancer element and a luciferase reporter gene, suggested MCD-1 cells are deficient in wild-type p53 and do not activate p53 on treatment with the anticancer agent adriamycin. The MCD-1 cell line is suggested to represent an abnormally differentiated cell type, which has some properties consistent with a multipotent neuronal phenotype while retaining some properties of immature cells of a glial lineage. The MCD-1 cell line can be used to provide a model of a medulloblastoma cell line that is resistant to growth-controlling and anticancer agents.

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

Similar content being viewed by others

References

  • Anderson, W. B. and Pastan, I. (1975) Altered adenylate cyclase activity: its role in growth regulation and malignant transformation of fibroblasts.Adv. Cyclic Nucleotide Res. 5, 681–698.

    PubMed  CAS  Google Scholar 

  • Bailey, P. and Cushing, H. (1925) Medulloblastoma cerebelli: a common type of midcerebellar tumor of childhood.Arch. Neurol. Psychiatry 14, 192–224.

    Google Scholar 

  • Berry, B. W., Boland, L. M., Hoch, D. B., and Dingledine, R. (1988)l-Glutamine binding site on N18-RE-105 neuroblastoma hybrid cells is not coupled to an ion channel.J. Neurochem. 51(4), 1176–1183.

    Article  PubMed  CAS  Google Scholar 

  • Bruce, J. H., Ramirez, A., and Lin, L. (1992) Effects of cyclic AMP and butyrate on cell cycle, DNA, RNA, and purine synthesis of cultured astrocytes.Neurochem. Res. 17, 315–320.

    Article  PubMed  CAS  Google Scholar 

  • Camins M. B., Cravioto H. M., Epstein F., and Ransohoff J. (1980) Medulloblastoma: An ultrastructural study—evidence for astrocytic and neuronal differentiation.Neurosurgery 6, 398–411.

    Article  PubMed  CAS  Google Scholar 

  • Clark W. C. and Bresseler J. (1988) Transforming growth factor-β-like activity in tumors of the central nervous system.J. Neurosurg. 68, 920–924.

    PubMed  CAS  Google Scholar 

  • Cogen, P. H., Daneshivar L., Metzger A. K., and Edwards M. S. (1990) Deletion mapping of the medulloblastoma locus on chromosome 17p.Genomics 8, 279–285.

    Article  PubMed  CAS  Google Scholar 

  • Currie D. N. and Dutton G. R. (1980) [3H] GABA uptake as a marker for cell type in primary cultures of cerebellum and olfactory bulb.Brain Res. 199, 473–481.

    Article  PubMed  CAS  Google Scholar 

  • Currie D. N. and Kelly J. S. (1981) Glial versus neuronal uptake of glutamate.J. Exp. Biol. 95, 181–193.

    PubMed  CAS  Google Scholar 

  • Dickson D. W., Hart M. N., Menezes A., and Cancilla P. A. (1983) Medulloblastomas with glial and rhabdomyoblastic differentiation.J. Neuropathol. Exp. Neurol. 42, 639–647.

    Article  PubMed  CAS  Google Scholar 

  • El-Diery W. S., Tokino T., Velculescu V. E., Levy D. B., Parsons R., Trent J. M., Lin D., Mercer E., Kinzler K. W., and Vogelstein B. (1993) WAF-1 a potential mediator of p53 tumor suppressor.Cell 75, 817–825.

    Article  Google Scholar 

  • El-Diery W. S., Harper J. W., O’Conner P. M., Velculescu V. E., Canman C. E., Jackman J., Pietenpol J. A., Burrell M., Hill D. E., Wang Y., Wiman K. G., Mercer E. W., Kastan M. B., Kohn K. W., Elledge S. J., Kinzler K. W., and Vogelstein B. (1994) WAF1/CIPI is induced in p53 mediated G1 arrest and apoptosis.Cancer Res. 54, 1169–1174.

    Google Scholar 

  • Friedman H. S., Burger P. C., Bigner S. H., Trojanowski J. Q., Wikstrand C. J., Halperin E. C., and Bigner D. D. (1985) Establishment and characterization of the human medulloblastoma cell line and transplantable xenograft D283 med.J. Neuropathol. Exp. Neurol. 44, 592–605.

    Article  PubMed  CAS  Google Scholar 

  • Friedman H. S., Burger P. C., Bigner S. H., Trojanowski J. Q. Brodeur G. M., He X., Wikstrand C. J., Kurtzburg J., Berens M. E., Halperin E. C., and Bigner D. D. (1988) Phenotypic and genotypic analysis of a human medulloblastoma cell line and transplantable xenograft (D341 MED) demonstrating amplification of c-myc.Am. J. Pathol. 130, 472–484.

    PubMed  CAS  Google Scholar 

  • Furukawa Y., Furukawa S., Omae H, Awatsuji H., and Hayashi, K. (1993) Alkylcatechols regulate NGF gene expression in astroglial cells via both protein kinase C- and cAMP independent mechanisms.J. Neurosci. Res. 35, 522–529.

    Article  PubMed  CAS  Google Scholar 

  • Ginty D. D., Glowacka D., Defranco C., and Wagner J. A. (1991) Nerve growth factor-induced neuronal differentiation after dominant repression of both type 1 and type 11 cAMP-dependent protein kinase activities.J. Biol. Chem. 266, 15,325–15,333.

    CAS  Google Scholar 

  • Goldman J. E. and Abramson B. (1990) Cyclic AMP-induced shape changes of astrocytes are accompanied by rapid depolymerization of actin.Brain Res. 528, 189–196.

    Article  PubMed  CAS  Google Scholar 

  • Gould V. E., Rorke L. B., Jansson D. S., Molenaar W. M., Trojanowski J. Q., Lee V. M. Y., Packer R. J., and Franke W. W. (1990b) Primitive neuroectodermal tumors of the central nervous system express neuroendocrine markers and may express all classes of intermediate filaments.Human Pathol. 21, 245–259.

    Article  CAS  Google Scholar 

  • Haglid K. G., Stavrou D., Ronnbuck L., Carlsson C. A., and Weidenbach W. (1973) The S-100 protein in water-soluble and pentanol-extractable form in normal human brain and tumors of the human nervous system. A quantitative study.J. Neurol. Sci. 20(1), 103–111.

    Article  PubMed  CAS  Google Scholar 

  • Haldman S. and McLennan H. (1973) The action of two inhibitors of glutamic acid uptake upon amino acid-induced and synaptic excitations of thalamic neurones.Brain Res. 63, 123–129.

    Article  Google Scholar 

  • Hayashi K., Motoi M., Nose S., Horie Y., Akagi T., Ogawa K., Taguchi K., Mizobuchi K., and Nishimoto, A. (1987) An immunohistochemical study on the distribution of glial fibrillary acidic protein, S-100 protein, neuron specific enolase, and neurofilament in medulloblastomas.Acta Pathol. Jpn. 37(1), 85–96.

    PubMed  CAS  Google Scholar 

  • He X, Skapek S. X., Wikstrand C. J., Friedman H. S., Trojanowski J. Q., Kemshead J. T., Coakham H. B., Bigner S. H., and Bigner D. D. (1989) Phenotypic analysis of four human medulloblastoma cell lines and transplantable xenografts.J. Neuropathol. Exp. Neurol. 48, 48–68.

    Article  PubMed  CAS  Google Scholar 

  • He X., Wikstrand C. J., Friedman H. S., and Bigner, S. H. (1991) Differentiation characteristics of newly established medulloblastoma cell lines (D384 Med, D425 Med, and D458 Med) and their transplantable xenografts.Lab Invest. 64, 833–843.

    PubMed  CAS  Google Scholar 

  • Herman M. M. and Rubinstein L. J. (1984) Divergent glial and neuronal differentiation in a cerebellar medulloblastoma in an organ culture system: in vitro occurrence of synaptic ribbons.Acta Neuropathol. 65, 10–24.

    Article  PubMed  CAS  Google Scholar 

  • Ishi D. N. (1978) Effect of tumor promoters on the response of cultured embryonic chick ganglia to nerve growth factor.Cancer Res. 38, 3886–3893.

    Google Scholar 

  • Jacobsen P. F., Jenkyn D. J., and Papadimiriou J. M. (1985) Establishment of a human cell line and its heterotransplantation into nude mice.J. Neuropathol. Exp. Neurol. 44, 5472–5485.

    Article  Google Scholar 

  • Jaros E., Lunee J., Perry P. J., Kelly P. J., and Pearson A. D. J. (1993) p53 protein overexpression identifies, a group of central primitive neuroectodermal tumors with poor prognosis.Br. J, Cancer 68, 801–807.

    CAS  Google Scholar 

  • Jennings M. T., Kaariainen I. T., Gold L., Maciunas R. J., and Commers P. A. (1994) TGβ1 and TGβ2 are potential growth regulators for medulloblastomas, primitive neuroectodermal tumors, and ependymomas: evidence in support of an autocrine hypothesis.Hum. Pathol. 25, 464–475.

    Article  PubMed  CAS  Google Scholar 

  • Kamamitopoulou E., Perentes E., and Diamantis I. (1993) p53 protein expression in central nervous system tumors: an immunohistochemical study with CM1 polyvalent and DO-7 monoclonal antibodies.Acta Neuropathol. 85, 611–616.

    Article  Google Scholar 

  • Keles G. E., Berger M. S., Lim R., Zaheer A., Denton A. L., and Silber J. R. (1992) Expression of glial fibrillary acidic protein in human medulloblastoma cells treated with recombinant glial maturation factor-β.Oncol. Res. 4, 431–437.

    PubMed  CAS  Google Scholar 

  • Keles G. E., Berger M. S., Schofield D., and Bothwell M. (1993) Nerve growth factor receptor expression in medulloblastomas and the potential role of nerve growth factor as a differentiating agent in medulloblastoma cell lines.Neurosurgery 32, 274–280.

    Article  PubMed  CAS  Google Scholar 

  • Kessler M., Baudry M., and Lynch G. (1987) Use of cystine to distinguish glutamate binding from glutamate sequestration.Neuroscience 81, 221–226.

    CAS  Google Scholar 

  • Kim U. H., Fink D., Kim H. S., Park D. J., Contreras M. L., Guroff G., and Rhee S. G. (1991) Nerve growth factor stimulates phosphorylation of phospholipase C-gamma in PC-12 cells.J. Biol. Chem. 266, 1359–1362.

    PubMed  CAS  Google Scholar 

  • Koyama Y., Tadashi I., and Hayata K. (1993) Endothelins modulate dibutyryl cAMP-induced stellation of cultured astrocytes.Brain Res. 600, 81–88.

    Article  PubMed  CAS  Google Scholar 

  • Kumanishi T., Washiyama K., Watabe K., and Sekiguchi K. (1985) Glial fibrillary acidic protein in medulloblastomas.Acta Neuropathol. (Berlin) 67, 1–5.

    Article  CAS  Google Scholar 

  • Levi G., Wilkin G. P., Ciotti M. T., and Johnstone G. P. (1983) Enrichment of differentiated, stellate astrocytes in cerebellar interneuron cultures as studied by GFAP immunofluorescence and autoradiographic uptake patterns with [3H]d-aspartate and [3H] GABA.Dev. Brain Res. 10, 227–24.

    Article  CAS  Google Scholar 

  • Loda M., Giangaspero, F. Badiali M., Capodieci P., and Pession, A. (1992) p53 Gene expression in medulloblastoma by quantitative polymerase chain reaction.Diagn. Mol. Pathol. 1, 36–44.

    Article  PubMed  CAS  Google Scholar 

  • Maria B. L., Steck P. A., Yung W. K., Milici A., Bruner J. M., Pathak S., and Becker F. F. (1989) The modulation of astrocytic differentiation in cells derived from medulloblastoma surgical specimen.J. Neurooncol. 7, 329–338.

    Article  PubMed  CAS  Google Scholar 

  • Maria B. L., Wong D., and Kalnins V. I. (1990) Dibutyryl cyclic AMP induces vimentin and GFAP expression in cultured medulloblastoma cells.Can. J. Neurol. Sci. 17, 15–20.

    PubMed  CAS  Google Scholar 

  • Michaelis E. K. (1979) The glutamate receptor-like protein of brain synaptic membranes is a metalloprotein.Biochem. Biophys Res. Comm. 87, 106–113.

    Article  PubMed  CAS  Google Scholar 

  • Pleasure S. J., Reddy U. R., Venkatakrishan G., Roy A. K., Chen J., Ross A. H., Trojanowski J. Q., Pleasure D. E., and Lee V. M. Y. (1990) Introduction of nerve growth factor (NGF) receptors into a medulloblastoma cell line results in expression of high and low affinity NGF receptors but not NGF-mediated differentiation.Proc. Natl. Acad. Sci. USA 87, 8496–8500.

    Article  PubMed  CAS  Google Scholar 

  • Poltorak M., Isono M., Freed W. J., Ronnett G. V., and Snyder S. H. (1992) Human cortical neuronal cell line (HCN-1): Further in vitro characterization and suitability for brain transplantation.Cell Transplantation 1, 3–15.

    PubMed  CAS  Google Scholar 

  • Quartin R. S., Cole C. N., Pipas J. M., Levine A. J. (1994) The amino-terminal functions of the simian virus 40 large T antigen are required to overcome wild-type p53-mediated growth arrest of cells.J. Virology 68, 1334–1341.

    PubMed  CAS  Google Scholar 

  • Roessman U., Velasco M. E., Gambetti P., and Autilio-Gambetti L. (1983) Neuronal and astrocytic differentiation in human neuroepithelial neoplasms.J. Neuropathol. Exp. Neurol. 42, 113–121.

    Article  Google Scholar 

  • Ronnett G. V., Hester L. D., Nye J. S., Conners K., and Snyder S. H. (1990) Human cortical neuronal cell line. Establishment from a patient with unilateral megalencephaly.Science 31, 603–605.

    Article  Google Scholar 

  • Schachner M. (1983)Immunohistochemistry and immunocytochemistry of neural cell types in vitro and in situ.Immunohistochemistry. Wiley-Interscience, New York.

    Google Scholar 

  • Shepard F. (1974) Cyclic AMP, cell growth, and the immune response: proceedings of the symposium held at Marco Island, FL, Jan. 8–10. Springer-Verlag, New York, pp. 112–119.

    Google Scholar 

  • Sporn M. B., Roberts A. B., Wakefield L. M., and de Crombrugghe B. (1987) Some recent advances in the chemistry and biology of transforming growth factor-beta.J. Cell Biol. 105, 1039–1045.

    Article  PubMed  CAS  Google Scholar 

  • Sutton L. N. and Packer R. J. (1994) Medulloblastomas, inPediatric Neurosurgery, 3rd ed. W. B. Saunders Co., Philadelpha, pp. 362–373.

    Google Scholar 

  • Tamura K., Shimizu K., Yamada M., Okamato Y., Matsui Y. M., Park K. C., Mabuchi E., Moriuchi S., and Mogami H. (1989) Expression of major histocompatibility complex on human medullolastoma cells with neuronal differentiation.Cancer Res. 49, 5380–5384.

    PubMed  CAS  Google Scholar 

  • Trojanowski J. Q. and Tohyama T. (1991) Molecular markers and new methods in neuropathology: central nervous system stem cell tumors.Curr. Opinion Neurol. Neurosurg. 4, 875–879.

    Google Scholar 

  • Trojanowski J. Q., Friedman H. S., Burger P. C., and Bigner D. D. (1987) A rapidly dividing human medulloblastoma cell line (D283 MED) expresses all three neurofilament subunits.Am. J. Pathol. 126, 358–363.

    PubMed  CAS  Google Scholar 

  • Trojanowski J. Q., Kelsten, M. L., and Lee V.M. Y. (1989) Phosphate dependent and independent neurofilament protein epitopes are expressed throughout the cell cycle in human medulloblastoma (D283 MED) cells.Am. J. Pathol. 135, 747–758.

    PubMed  CAS  Google Scholar 

  • Trojanowski J. Q., Tohyama T., and Lee M. Y. (1992) Medulloblastomas and related primitive neuroectodermal brain tumors of childhood recapitulate molecular milestones in the maturation of neuroblasts.Mol. Chem. Neuropathol. 17, 121–135.

    Article  PubMed  CAS  Google Scholar 

  • Velasco M. E., Ghobrial M. W., and Ross E. R. (1985) Neuron specific enolase and neurofilament protein as markers of differentiation in medulloblastoma.Surg. Neurol. 23, 177–182.

    Article  PubMed  CAS  Google Scholar 

  • Vogelstein B. and Kinzler K. W. (1992) p53 Function and dysfunction.Cell 70, 523–526.

    Article  PubMed  CAS  Google Scholar 

  • Westlund K. N., Lu Y., Werrbach-Perez K., Huselbosch C. E., Morgan B., Pizzo D. P., Eisenberg H. M., and Perez-Polo J. R. (1992) Effects of nerve growth factor and acetyl-l-carnitine arginyl amide on the human neuronal line HCN-1A.Int. J. Dev. Neurosci. 10, 361–373.

    Article  PubMed  CAS  Google Scholar 

  • Wilkin G. P., Levi G., Johnstone S. R., and Riddle P. N. (1983) Cerebellar astroglial cells in primary culture: expression of different morphological appearances and different ability to take up [3H]d-aspartate and [3H] GABA.Dev. Brain Res. 10, 265–277.

    Article  CAS  Google Scholar 

  • Wu J. K., Ye Z., and Darras, B. T. (1993) Frequency of p53 tumor suppressor gene mutations in human primary brain tumors.Neurosurgery 33, 824–830.

    Article  PubMed  CAS  Google Scholar 

  • Wyllie F. S., Dawson T. Bond J. A., Goretzki P., Game, S., Prime S., and Wynford-Thomas D. (1991) Correlated abnormalities of transforming growth factor-β1 response and p53 expression in thyroid epithelial cell transformation.Mol. Cell Endocrinol. 76, 13–21.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Section on Preclinical Neuroscience, Neuropsychiatry Branch, NIMH Neuroscience Center at St. Elizabeths, 2700 Martin Luther King Ave, 20032, Washington, DC

    Kraig D. Moore, Ora Dillon-Carter, Concepcion Conejero, Maciej Poltorak & William J. Freed

  2. Genesys Neurosurgical Associates, Flint, MI

    Michael Chedid

  3. Laboratory of Molecular Medicine and Neuroscience, NINDS, Bethesda, MD

    Carlo Tornatore

Authors
  1. Kraig D. Moore
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ora Dillon-Carter
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Concepcion Conejero
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maciej Poltorak
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael Chedid
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Carlo Tornatore
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. William J. Freed
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Moore, K.D., Dillon-Carter, O., Conejero, C. et al. In vitro properties of a newly established medulloblastoma cell line, MCD-1. Molecular and Chemical Neuropathology 29, 107–126 (1996). https://doi.org/10.1007/BF02814996

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02814996

Index Entries

Search

Navigation