Skip to main content
SpringerLink
Log in

Mitogenesis in Glioblastoma Multiforme Cell Lines: A Role for NGF and its TrkA Receptors

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

Abstract

Neurotrophins have definitive roles in the growth/maintenance of neuronal populations, but their function in malignant gliomas is unknown. The ability for nerve growth factor (NGF) to serve as a mitogenic agent was investigated in several human glioblastoma multiforme (GBM) cell lines, including U251, U87, and U373. In a serum-free medium, the addition of NGF (200 ng/ml) to these cell lines increased cell counts over controls, after 3 days in culture by 9%, 16%, and 33%, respectively. Dose-dependent increases in cell counts and [3H]thymidine uptake were found in the more rigorously investigated U373 cell line. Proteins for both the high affinity NGF-specific tyrosine kinase binding site (p140TrkA; TrkA) and the low affinity neurotrophin (p75NTR) receptor were present in all three GBM cell lines. TrkA mRNA was identified in U373 (only cell line studied). NGF-stimulated proliferation was inhibited in a dose-dependent fashion by K252a, a blocker of Trk-induced receptor kinases. NGF, measured by ELISA, was detectable in all GBM cell lines even after 7 days of growth in serum-free medium. These data suggest that GBM cell growth can be enhanced by NGF acting via Trk receptor phosphorylation. Future studies of antiproliferative therapies should consider agents directed against intracellular Trk signaling cascades.

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

  1. Chao MV: Nerve growth factor. In: Sporn MB, Roberts AB (eds) Peptide Growth Factors and Their Receptors II. Springer Publishing Co, New York, 1991, pp 135–165

    Google Scholar 

  2. Hutton LA, de Vellis J, Perez-Polo Jr: Expression of p75NGFR, trkA, and trkB mRNA in rat C6 glioma and type I astrocyte cultures. J Neurosci Res 32: 375–383, 1992

    Google Scholar 

  3. Lindsay RM, Wiegand SJ, Altar CA, DiStefano PS: Neurotrophic factors: from molecule to man. Trends Neurosci 17: 182–190, 1994

    Google Scholar 

  4. Kumar S, Peña LA, de Vellis J: CNS glial cells express neurotrophin receptors whose levels are regulated by NGF. Mol Brain Res 17: 163–168, 1993

    Google Scholar 

  5. Foschini DR, Kestler AM, Egger MD, Crockett DP: The up-regulation of trkA and trkB in dorsal column astrocytes following dorsal rhizotomy. Neurosci Lett 169: 21–24, 1994

    Google Scholar 

  6. Condorelli DF, Dell'Albani P, Mudo G, Timmusk T, Bellvardo N: Expression of neurotrophins and their receptors in primary astroglial cultures: induction by cyclic AMPelevating agents. J Neurochem 63: 509–516, 1994

    Google Scholar 

  7. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  8. Karp CL, El-Safi SH, Wynn TA, Satti MM, Kordofani AM, Hashim FA, Hag-Ali M, Neva FA, Nutman TB, Sacks DL: In vivo cytokine profiles in patients with kala-azar: marked elevation of both interleukin-10 and interferon-gamma. J Clin Invest 91: 1644–1648, 1993

    Google Scholar 

  9. Martin-Zanca D, Oskam R, Mitra G, Copeland T, Barbacid M: Molecular and biochemical characterization of the human trk proto-oncogene. Mol Cell Biol 9: 24–33, 1989

    Google Scholar 

  10. Barker FG, Israel MA: The molecular biology of brain tumors. Neurologic Clinics 13: 701–721, 1995

    Google Scholar 

  11. Marushige Y, Raju NR, Marushige K, Koestner A: Modulation of growth and morphological characteristics in glioma cells by nerve growth factor and glial maturation factor. Cancer Res 47: 4109–4115, 1987

    Google Scholar 

  12. Rakowicz-Szulczynska EM, Koprowski H: Antagonistic effect of PDGF and NGF on transcription of ribosomal DNA and tumor cell proliferation. Biochem Biophys Res Commun 163: 649–656, 1989

    Google Scholar 

  13. Lüdecke G, Unsicker K: Mitogenic effect of neurotrophic factors on human IMR 32 neuroblastoma cells. Cancer 65: 2270–2278, 1990

    Google Scholar 

  14. Oelmann E, Sreter L, Schuller I, Serve H, Koenigsmann M, Wiedenmann B, Oberberg D, Reufi B, Thiel E, Berdel WE: Nerve growth factor stimulates clonal growth of human lung cancer cell lines and a human glioblastoma cell line expressing high-affinity nerve growth factor binding sites involving tyrosine kinase signaling. Cancer Res 55: 2212–2219, 1995

    Google Scholar 

  15. Ikeda T, Puro DG: Nerve growth factor: a mitogenic signal for retinal Müller glial cells. Brain Res 649: 260–264, 1994

    Google Scholar 

  16. Vinores SA, Koestner A: The effect of nerve growth factor on undifferentiated glioma cells. Cancer Letters 10: 309–318, 1980

    Google Scholar 

  17. Mesner PW, Winters TR, Green SH: Nerve growth factor withdrawal-induced cell death in neuronal PC12 cells resembles that in sympathetic neurons. J Cell Biol 6: 1669–1680, 1992

    Google Scholar 

  18. Junghanel A, Berndt A, Kosmehl H, Hyckel D: Cell kinetics of oral squamous epithelial carcinomas. Determination of the proliferation index and apoptosis rate with the TUNEL method. Mund-, Kiefer-Und Gesichtschirurgie 2: 250–255, 1998

    Google Scholar 

  19. Lee KF, Davies AM, Jaenisch R: p75 deficient embryonic dorsal root sensory and neonatal sympathetic neurons display a decreased sensitivity to NGF. Development 120: 1027–1033, 1994

    Google Scholar 

  20. Chao MV, Hempstead BL: p75 and Trk: a two receptor system. Trends Neurosci 18: 321–326, 1995

    Google Scholar 

  21. Taglialatatela G, Hibbert CJ, Hutton LA, Werrbach-Perez K, Perez-Polo JR: Suppression of p140trkA does not abolish nerve growth factor-mediated rescue of serum-free PC12 cells. J Neurochem 66: 1826–1835, 1996

    Google Scholar 

  22. Bredesen DE, Rabizadeh S: p75NTR and apoptosis: Trkdependent and Trk-independent effect. Trends Neurosci 20: 287–290, 1997

    Google Scholar 

  23. Cortazzo MH, Kassis ES, Sproul KA, Schor NF: Nerve growth factor (NGF)-mediated protection of neural crest cells from antimitotic agent-induced apoptosis: the role of the low-affinity NGF receptor. J Neurosci 16: 3895–3899, 1996

    Google Scholar 

  24. Tolkovsky A: Neurotrophic factors in action-new dogs and new tricks. Trends Neurosci 20: 1–3, 1997

    Google Scholar 

  25. Blumberg D, Radeke MJ, Feinstein SC: Specificity of nerve growth factor signaling: differential patterns of early tyrosine phosphorylation events induced by NGF, EGI, and bFGF. J Neurosci Res 41: 628–639, 1995

    Google Scholar 

  26. Russell DS: Neutrophins: mechanisms of action. The Neuroscientist 1: 3–6, 1995

    Google Scholar 

  27. Heumann R: Neurotrophin signaling. Curr Opin Neurobiol 4: 668–679, 1994

    Google Scholar 

  28. Nye SH, Squinto SP, Glass DJ, Stitt TN, Hantzopoulos P, Macchi MJ, Lindsay NS, Ip NY, Yancopoulos GD: K-252a and staurosporine selectively block autophosphorylation of neurotrophin receptors and neurotrophin-mediated responses. Mol Biol Cell 3: 677–686, 1992

    Google Scholar 

  29. Greene LA, Kaplan DR: Early events in neurotrophin signaling via Trk and p75 receptors. Curr Opin Neurobiol 5: 579–587, 1995

    Google Scholar 

  30. Kaplan DR: Signal transduction by Trk receptors. In: Iba?ez CF, Hokfelt T, Olsow L, Fuxe K, Jornvall M, Ottoson L (eds) Life and Death in the Nervous System. Pergamon, Oxford, 1995, pp 37–54

    Google Scholar 

  31. Ohmichi M, Decker SJ, Pang L, Saltiel AR: Inhibition of the cellular actions of nerve growth factor by staurosporine and K252a results from attenuation of the activity of the trk tyrosine kinase. Biochem 31: 4034–4039, 1992

    Google Scholar 

  32. Tapley P, Lamballe F, Barbacid M: K252a is a selective inhibitor of the tyrosine protein kinase activity of the trk family of oncogenes and neurotrophin receptors. Oncogene 7: 371–381, 1992

    Google Scholar 

  33. Koizumi S, Contreras ML, Matsuda Y, Hanna T, Lazarovici P, Guroff G: K-252a: a specific inhibitor of the action of nerve growth factor on PC12 cells. J Neurosci 8: 715–721, 1988

    Google Scholar 

  34. Koizumi S, Mutoh T, Ryazanov A, Rudkin BB, Guroff G: Nerve growth factor-sensitive phosphorylations and the action of K-252a on PC12 cells. In: Horrocks LA, Neff NH, Yates AJ, Hadjiconstantinou M (eds) Trophic Factors and the Nervous System. Raven Press, New York, 1990, pp 195–202

    Google Scholar 

  35. Becker D, Meier CD, Herlyn M: Proliferation of human malignant melanomas is inhibited by antisense oligodeoxynucleotides targeted against basic fibroblast growth factor. EMBO J 8: 3685–3691, 1989

    Google Scholar 

  36. Morrison RS: Suppression of basic fibroblast growth factor expression by antisense oligodeoxynucleotides inhibits the growth of transformed human astrocytes. J Biol Chem 266: 728–734, 1991

    Google Scholar 

  37. Spranger M, Lindholm D, Bandtlow C, Heumann R, Gnahn H, Näher-Noé M, Thoenen H: Regulation of nerve growth factor (NGF) synthesis in the rat central nervous system: comparison between the effects of interleukin-1 and various growth factors in astrocyte cultures and in vivo. Eur J Neurosci 2: 69–76, 1990

    Google Scholar 

  38. Yoshida K, Gage FH: Cooperative regulation of nerve growth factor synthesis and secretion in fibroblasts and astrocytes by fibroblast growth factor and other cytokines. Brain Res 569: 14–25, 1992

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurology, USA

    Harvey S. Singer, Bendt Hansen, Daniel Martinie & Christopher L. Karp

  2. Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Harvey S. Singer

  3. Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Christopher L. Karp

Authors
  1. Harvey S. Singer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bendt Hansen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Daniel Martinie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christopher L. Karp
    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

Singer, H.S., Hansen, B., Martinie, D. et al. Mitogenesis in Glioblastoma Multiforme Cell Lines: A Role for NGF and its TrkA Receptors. J Neurooncol 45, 1–8 (1999). https://doi.org/10.1023/A:1006323523437

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006323523437

Search

Navigation