Advertisement

Clinical & Experimental Metastasis

, Volume 12, Issue 6, pp 405–415 | Cite as

The role of extracellular matrix in human astrocytoma migration and proliferation studied in a microliter scale assay

  • Michael E. Berens
  • Monique D. Rief
  • Melinda A. Loo
  • Alf Giese
Research Papers

Abstract

Ligands in the extracellular matrix (ECM) are known to mediate migration of normal as well as tumor cells via adhesion molecules such as the integrin receptor family. We developed a microliter scale (15-20futotal volume) monolayer migration assay to investigate the ability of astrocytoma cells to disperse on surfaces coated with purified human ECM protein ligands. In this system the rate of radial migration of the cell population was constant over time. For human astrocytoma cell lines U-251 and SF-767, laminin and collagen type IV supported a migratory phenotype; fibronectin and vitronectin only minimally supported migration. The different ECM proteins also influenced growth rate: cells on laminin and collagen had a protracted lag phase. Furthermore, migrating cells seeded on laminin or collagen showed a lower labeling index than did stationary cells in the central, crowded region on the same substrate. This micro-scale migration assay should enable detailed molecular and biochemical studies of the determinants of migration.

Keywords

astrocytoma extracellular matrix integrins migration proliferation 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Müller W, Afra D and Schröder R, 1977, Supratentorial recurrence of gliomas: morphological studies in relation to time intervals with astrocytomas.Acta Neurochirugia (Wien),37, 75–91.Google Scholar
  2. 2.
    Burger PC, Heinz ER, Shibata T and Kleihues P, 1988, Topographic anatomy and CT correlations in the untreated glioblastoma multiforme.J Neurosurg,68, 698–704.Google Scholar
  3. 3.
    Liotta LA and Stetler-Stevenson WG, 1991, Tumor invasion and metastasis: an imbalance of positive and negative regulation.Cancer Res,51, 5054s-59s.Google Scholar
  4. 4.
    Hynes RO, 1987, Integrins: a family of cell surface receptors.Cell,48, 549–57.Google Scholar
  5. 5.
    Hynes RO, 1992, Integrins: Versatility, modulation, and signaling in cell adhesion.Cell,69, 11–25.Google Scholar
  6. 6.
    Wewer UM, Taraboletti G, Sobel ME, Albrechtsen R and Liotta LA, 1987, Role of laminin receptor in tumor cell migration.Cancer Res,47, 5691–8.Google Scholar
  7. 7.
    McCarthy JB, Basara ML, Palm SL, Sas DF and Furcht LT, 1985, The role of cell adhesion proteins laminin and fibronectin in the movement of malignant and metastatic cells.Cancer Met Rev,4, 125–52.Google Scholar
  8. 8.
    Carbonetto S, 1984, The extracellular matrix of the nervous system.Trends Neurol Sci,7, 382–7.Google Scholar
  9. 9.
    Rutka JT, Apodaca G, Stern R and Rosenblum M, 1988, The extracellular matrix of the central and peripheral nervous system: structure and function.J Neurosurg,69, 155–70.Google Scholar
  10. 10.
    Venstrom KA and Reichard LF, 1993, Extracellular matrix 2: Role of extracellular matrix molecules and their receptors in the nervous system.FASEB J,7, 996–1003.Google Scholar
  11. 11.
    Bjerkvig R, Learum OD and Mella O, 1986, Glioma cell interaction with fetal rat brain aggregatesin vitro and with brain tissuein vivo.Cancer Res,46, 4071–9.Google Scholar
  12. 12.
    Scherer HJ, 1940, The forms of growth in gliomas and their practical significance.Brain,63, 1–35.Google Scholar
  13. 13.
    Schiffer D, 1986, Neuropathology and imaging. The ways in which glioma spreads and varies in its histological aspect. In: Walker MD, Thomas DGT, eds.Biology of Brain Tumour. Boston: Martinus Nijhoff, 163–72.Google Scholar
  14. 14.
    Jones TR, Ruoslahti E, Schold SC and Bigner DD, 1982, Fibronectin and glial fibrillary acidic protein expression in normal human brain and anaplastic human gliomas.Cancer Res,42, 299–305.Google Scholar
  15. 15.
    Alitalo K, Bornstein P, Vaheri A and Sage H, 1983, Biosynthesis of an unusual collagen type by human astrocytoma cellsin vitro.J Biol Chem,258, 2653–61.Google Scholar
  16. 16.
    McKeever PE, Fligiel SE, Varani J, Castle RJ and Hood TW, 1989, Products of cells cultured from gliomas. VII. Extracellular matrix proteins of gliomas which contain glial fibrillary acidic protein.Lab Invest,60, 286–95.Google Scholar
  17. 17.
    Liesi P, Dahl D and Vaheri A, 1983, Laminin is produced by early rat astrocytes in primary culture.J Cell Biol,96, 920–24.Google Scholar
  18. 18.
    Liesi P, Kirkwood T and Vaheri A, 1986, Fibronectin is expressed by astrocytes cultured from embryonic and early postnatal rat brain.Exp Cell Res,163, 175–85.Google Scholar
  19. 19.
    McComb RD and Bigner DD, 1985, Immunolocalisation of laminin in neoplasms of the central and peripheral nervous systems.J Neuropath Exp Neurol,44, 242–53.Google Scholar
  20. 20.
    Varani J, Orr W and Ward PA, 1978, A comparison of the migration patterns of normal and malignant cells in two assay systems.Am J Pathol,90, 159–71.Google Scholar
  21. 21.
    Berens ME, Weisman AS, Spencer DR,et al. 1991, Growth properties and oncogene expression in 2 newly derived glioma cell lines: assessment of growth determinants.Proc AACR,31, 273.Google Scholar
  22. 22.
    Bigner DD, Bigner SH and Ponten, J 1981, Heterogeneity of genotypic and phenotypic characteristics of 15 permanent cell lines derived from human gliomas.J Neuropath Exp Neurol,40, 201–15.Google Scholar
  23. 23.
    Kramer R, McDonald KA, Crowley E, Ramos D and Damsky CH, 1989, Integrin-related complexes mediate the adhesion of melanoma cells to basement membrane.Cancer Res,49, 393–402.Google Scholar
  24. 24.
    Vuento M and Vaheri A, 1979, Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions.Biochem J,183, 331–7.Google Scholar
  25. 25.
    Giese A, Rief MD, Loo MA and Berens ME, 1994, Determinants of human astrocytoma migration.Cancer Res,54, 3897–904.Google Scholar
  26. 26.
    Phillips DR, Charo IF and Scarborough RM, 1991, GPIIb-IIIa: the responsive integrin.Cell,65, 359–62.Google Scholar
  27. 27.
    Tawil N, Wilson P and Carbonetto S, 1993, Integrins in point contacts mediate cell spreading: factors that regulate integrin accumulation in point contacts vs focal contacts.J Cell Biol,120, 261–71.Google Scholar
  28. 28.
    Varani J, Carey TE, Fligiel SE, McKeever PE and Dixit V, 1987, Tumor type-specific differences in cell-substrate adhesion among human tumor cell lines.Int J Cancer,39, 397–403.Google Scholar
  29. 29.
    Romualdez AG Jr and Ward PA, 1975, A unique complement derived chemotactic factor for tumor cells.Proc Nat Acad Sci,72, 4128–32.Google Scholar
  30. 30.
    Repesh LA, 1989, A newin vitro assay for quantitating tumor cell invasion.Inv Met,9, 192–208.Google Scholar
  31. 31.
    Hendrix MJ, Gehlsen KR, Wagner HN Jr,et al. 1985,In vitro quantification of melanoma tumor cell invasion.Clin Exp Metastasis,3, 221–33.Google Scholar
  32. 32.
    Thomas LA and Yamada KM, 1992, Contact stimulation of cell migration.J Cell Science,103, 1211–14.Google Scholar
  33. 33.
    Henderson E, Haydon PG and Sakaguchi DS, 1992, Actin filament dynamics in living glial cells imaged by atomic force microscopy.Science,257, 1944–6.Google Scholar
  34. 34.
    Woods A, McCarthy JB, Furcht LT and Couchman JR, 1993, A synthetic peptide from the COOH-terminal heparin-binding domain of fibronectin promotes focal adhesion formation.Mol Biol Cell,4, 605–13.Google Scholar

Copyright information

© Rapid Communications of Oxford Ltd 1994

Authors and Affiliations

  • Michael E. Berens
    • 1
  • Monique D. Rief
    • 1
  • Melinda A. Loo
    • 1
  • Alf Giese
    • 1
  1. 1.Neuro-Oncology LaboratoryBarrow Neurological Institute of Saint Joseph's Hospital and Medical CenterPhoenixUSA

Personalised recommendations