Skip to main content
SpringerLink
Log in

GFAP expression in the subcutaneous tumors of immature glial cell line (HITS glioma) derived from ENU-induced rat glioma

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

Summary

In order to evaluate the proliferation and differentiation potentials of ethylnitrosourea (ENU)-induced rat glioma cells, the authors attempted to obtain a cell line that maintains glial features in long-term culture. One of five cell lines cultivated from ENU-induced rat gliomas merited particular interest because of the differentiation of its neoplastic glia. This cell line, designated as HITS glioma, had a polygonal cell body and formed a monolayer with pile-up fociin vitro, in contrast to the other cell lines, which displayed a mesenchymal change through passages. GFAP-positive cells, found in the primary culture, disappeared in the late passages of HITS glioma, as they did in the other cell lines. Galactocerebroside (GC), GD3 ganglioside, and Leu7 were not expressed in the cell lines during culture. Subcutaneous inoculation of HITS glioma into neonatal rats induced tumors with histopathological components mimicking the histopathological appearance of ENU-induced gliomas. The components also had a fraction of GFAP-positive cells. Such findings indicate that HITS glioma cells may be composed of immature glial cells which are able to differentiate into astrocytic cells under certain conditions.

Several growth factors which play a role in gliogenesis were used to evaluate the mechanism(s) of proliferation and/or differentiation of HITS glioma. These growth factors did not induce the expression of GFAP and other antigenic expression in HITS glioma, even though some promoted the proliferation of HITS glioma. Although the mechanism involving the astrocytic differentiation of HITS glioma is unknown, HITS glioma may serve as an effective tool in research to evaluate the mechanisms of proliferation and differentiation of neoplastic glia.

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. Bogler O, Wren D, Barnett SC, Land H, Noble M: Cooperation between two growth factors promotes extended selfrenewal and inhibits differentiation of oligodendrocytetype-2 astrocyte (O-2A) progenitor cells. Proc Natl Acad Sci USA 87: 6368–6372, 1990

    PubMed  Google Scholar 

  2. Borit A, McIntosh GC: Myelin basic protein and glial fibrillary acidic protein in human fetal brain. Neuropathol Appl Neurobiol 7: 279–287, 1981

    PubMed  Google Scholar 

  3. Conley FK: The immunocytochemical localization of GFA Protein in experimental murine CNS tumors. Acta Neuropathol (Berl) 45: 9–16, 1979

    Google Scholar 

  4. Dahl D: The vimentin-GFA Protein transition in rat neuroglia cytoskeleton occurs at the time of myelination. J Neurosci Res 6: 741–748, 1981

    PubMed  Google Scholar 

  5. de la Monte SM: Uniform lineage of oligodendrogliomas. Am J Pathol 135(3): 529–540, 1989

    PubMed  Google Scholar 

  6. Druckrey H, Ivankovic S, Preussmann R: Teratogenic and carcinogenic effects in the offspring after single injection of ethylnitrosourea to pregnant rats. Nature 210: 1378–1379, 1966

    PubMed  Google Scholar 

  7. Dutly F, Schwab ME: Neurons and astrocytes influence the development of purified O-2A progenitor cells. Glia 4: 559–571, 1991

    PubMed  Google Scholar 

  8. Gammeltoft S, Ballotti R, Kowalski A, Westermark B, Obberghan EV: Expression of two types of receptor for insulin-like growth factors in human malignant glioma. Cancer Res 48: 1233–1237, 1988

    PubMed  Google Scholar 

  9. Giordana MT, Mauro A, Schiffer D: Glycosaminoglycans of brain tumors transplacentally induced by ENU in the rat. Acta Neuropathol (Berl) Suppl VII: 79–80, 1981

    Google Scholar 

  10. Giulian D, Young DG, Woodward J, Brown DC, Lachman LB: Interleukin-I is an astroglial growth factor in the developing brain. J Neurosci 8: 709–714, 1988

    PubMed  Google Scholar 

  11. Gratzner HG: Monoclonal antibody to 5-bromo and 5-iododeoxyuridine: A new reagent for detection of DNA replication. Science 218: 474–475, 1982

    PubMed  Google Scholar 

  12. Han VKM, Lauder JM, D'Ercole AJ: Characterization of Somatomedin/Insulin-like growth factor receptors and correlation with biologic action in cultured neonatal rat astroglial cells. J Neurosci 7(2): 501–511, 1987

    PubMed  Google Scholar 

  13. Herpers MJHM, Budka H: Glial fibrillary acidic protein (GFAP) in oligodendroglial tumors: Gliofibrillary oligodendroglioma and transitional oligoastrocytoma as subtypes of oligodendroglioma. Acta Neuropathol (Berl) 64: 265–272, 1984

    Google Scholar 

  14. Higami Y: Response of ethylnitrosourea-induced rat glioma to growth factors in primary culture. Nagasaki Med J 62(2): 150–172, 1990 (in Japanese with English abstract)

    Google Scholar 

  15. Hsu S-M, Raine L, Fanger H: Use of avidin-biotin-peroxydase complex (ABC) in immunoperoxydase techniques: A comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29(4): 577–580, 1981

    PubMed  Google Scholar 

  16. Jagadha V, Halliday WC, Becker LE: Glial fibrillary acidic protein (GFAP) in oligodendrogliomas: A reflection of transient GFAP expression by immature oligodendroglia. Can J Neurol Sci 13: 307–311, 1986

    PubMed  Google Scholar 

  17. Kiess W, Lee L, Graham DE, Greenstein L, Tseng LY-H, Rechler MM, Nissley SP: Rat C6 glial cells synthesize Insulin-like growth factor I (IGF-I) and express IGF-I receptors and IGF-II/mannose 6-phosphate receptors. Endocrinology 124: 1727–1736, 1989

    PubMed  Google Scholar 

  18. Kim SU, Moretto G, Shin DH, Lee VM: Modulation of antigenic expression in cultured adult human oligodendrocytes by derivatives of adenosine 3′, 5′-cyclic monophosphate. J Neurol Sci 69: 81–91, 1985

    PubMed  Google Scholar 

  19. Ko L, Koestner A, Wechsler W: Morphological characterization of nitrosourea-induced glioma cell lines and clones. Acta Neuropathol (Berl) 51: 23–31, 1980

    Google Scholar 

  20. Koestner A, Swenberg JA, Wechsler W: Transplacental production with ethylnitrosourea of neoplasms of the nervous system in Sprague-Dawley rats. Am J Pathol 63(1): 37–50, 1971

    PubMed  Google Scholar 

  21. Le Vine SM, Goldman JE: Ultrastructural characteristics of GD3 ganglioside-positive immature glia in rat forebrain white matter. J Comp Neurol 277: 456–464, 1988

    PubMed  Google Scholar 

  22. Mauro A, Giordana MT, Migheli A, Schiffer D: Glial fibrillary acidic protein (GFAP) in rat brain tumors transplacentally induced by ethylnitrosourea (ENU). J Neurol Sci 61: 349–355, 1983

    PubMed  Google Scholar 

  23. McKeever PE: Functional aspects of alterations in glioma cell antigens. Neuroimmunol Res 2: 19–39, 1989

    Google Scholar 

  24. McMorris FA, Smith TM, DeSalvo S, Furlanetto RW: Insulin-like growth factor I/Somatomedin C: A potent inducer of oligodendrocyte development. Proc Natl Acad Sci USA 83: 822–826, 1986

    PubMed  Google Scholar 

  25. McMorris FA, Dubois-Dalcq M: Insulin-like growth factor I promotes cell proliferation and oligodendroglial commitment in rat glial progenitor cells developingin vitro. J Neurosci Res 21: 199–209, 1988

    PubMed  Google Scholar 

  26. Merrill MJ, Edwards NA: Insulin-like growth factor-I receptors in human glial tumors. J Clin Endocrinol Metab 71(1): 199–209, 1990

    PubMed  Google Scholar 

  27. Nakagawa Y, Perentes E, Rubinstein LJ: Immunohistochemical characterization of oligodendrogliomas: an analysis of multiple markers. Acta Neuropathol (Berl) 72: 15–22, 1986

    Google Scholar 

  28. Peatau A: Glial fibrillary acidic protein, vimentin and fibronectin in primary cultures of human glioma and fetal brain. Acta Neuropathol (Berl) 75: 448–455, 1988

    Google Scholar 

  29. Raff MC, Rhona M, Fields KL, Lisak RP, Dorfman SH, Silberberg DH, Gregson NA, Leibowitz S, Kennedy MC: Galactocerebroside is a specific cell-surface antigenic marker for oligodendrocytes in culture. Nature 274: 813–816, 1978

    PubMed  Google Scholar 

  30. Raff MC, Miller RH, Noble M: A glial progenitor cell that developsin vitro into an astrocyte or an oligodendrocyte depending on culture medium. Nature 303(2): 390–396, 1983

    PubMed  Google Scholar 

  31. Reifenberger G, Bilzer T, Seitz RJ, Wechsler W: Expression of vimentin and glial fibrillary acidic protein in ethylnitrosourea-induced rat gliomas and glioma cell lines. Acta Neuropathol (Berl) 78: 270–282, 1989

    Google Scholar 

  32. Ross R, Raines EW, Bowen-Pope DF: The biology of platelet-derived growth factor. Cell 46: 155–169, 1986

    PubMed  Google Scholar 

  33. Sandberg A-C, Engberg C, Lake M, von Holst H, Sara VR: The expression of insulin-like growth factor I and insulin-like growth factor II genes in the human fetal and adult brain and in glioma. Neurosci Lett 93: 114–119, 1988

    PubMed  Google Scholar 

  34. Schechter AL, Stern DF, Vaidyanathan L, Decker SJ, Drebin JA, Green MI, Weinberg RA: The neu oncogene: an erbB-related gene encoding a 185, 000-Mr tumour antigen. Nature 312: 513–516, 1984

    PubMed  Google Scholar 

  35. Shapiro JR: Biology of gliomas: Heterogeneity, oncogenes, growth factors. Semin Oncol 13(1): 4–15, 1986

    PubMed  Google Scholar 

  36. Shemer J, Raizada MK, Masters BA, Ota A, LeRoith D: Insulin-like growth factor I receptors in neuronal and glial cells. J Biol Chem 262(16): 7693–7699, 1987

    PubMed  Google Scholar 

  37. Shimokawa I: Immunohistochemical properties of glial cells of ethylnitrosourea induced brain tumor in the rat. Acta Med Nagasaki 31: 90–116, 1986

    Google Scholar 

  38. Sternberger NH, Itoyama Y, Kies MW, Webster HF: Myelin basic protein demonstrated immunocytochemically in oligodendroglioma prior to myelin sheath formation. Proc Natl Acad Sci USA 75(5): 2521–2524, 1978

    PubMed  Google Scholar 

  39. Takesima F, Iwasaki K, Shimokawa I, Ikeda T, Matsuo T: Immunohistochemical localization of gangliosides in ENU-induced rat glioma. Acta pathol Jpn 42(8): 558–565, 1992

    PubMed  Google Scholar 

  40. Yoshimine T, Maruno M, Ushio Y, Hayakawa T, Nakajima Y, Mogami H: Intermediate filaments and anaplastic change of ENU-induced gliomas: immunohistochemical study with vimentin and astroprotein (GFAP). J Neurooncol 5: 377–385, 1987

    PubMed  Google Scholar 

  41. Yoshino T, Motoi M, Ogawa K: Morphological maturation of tumor cells induced by ethylnitrosourea (ENU) in rat brains. I. On the tumors by administration of ENU in the late gestational stage. Acta Pathol Jpn 35(6): 1385–1396, 1985

    PubMed  Google Scholar 

  42. Zapf J, Froesch ER, Humbel RE: The Insulin-like growth factors (IGF) of human serum: Chemical and biological characterization and aspects of their possible physiological role. Curr Top Cell Regul 19: 257–309, 1981

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The First Department of Pathology, Nagasaki University School of Medicine, 1-12-4 Sakamoto, 852, Nagasaki, Japan

    Yuji Ikeno, Isao Shimokawa, Yoshikazu Higami & Takayoshi Ikeda

Authors
  1. Yuji Ikeno
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Isao Shimokawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoshikazu Higami
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takayoshi Ikeda
    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

Ikeno, Y., Shimokawa, I., Higami, Y. et al. GFAP expression in the subcutaneous tumors of immature glial cell line (HITS glioma) derived from ENU-induced rat glioma. J Neuro-Oncol 17, 191–204 (1993). https://doi.org/10.1007/BF01049975

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01049975

Key words

Search

Navigation