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Relationship of the demonstration of intermediate filament protein to kinetics of three human neuroepithelial tumor cell lines. Lack of neural-related proteins in most cells in S phase: a double-labeled immunohistochemical study on matrix cultures.

Abstract
The immunocytochemical demonstration of intermediate filament proteins in three human neuroepithelial tumor cell lines maintained in vitro on a three-dimensional matrix was correlated with the proportion of cells in S phase. The cell lines of a medulloblastoma (D283 Med), a retinoblastoma (WERI-Rb1), and an astrocytic glioma (U-251 MG) were cultured in an organ culture system, pulse-fed with bromodeoxyuridine, and double-labeled by immunoperoxidase and by the avidin-biotin peroxidase complex method for bromodeoxyuridine and for intermediate filament proteins [each triplet of neurofilament proteins, as well as vimentin and glial fibrillary acidic (GFA) protein] using eight different antibodies. The average percentages of bromodeoxyuridine-labeled cells for the D283 Med, WERI-Rb1, and U-251 MG lines were respectively, 25, 32, and 12% 30 minutes after pulse labeling. In the D283 Med line, 15- greater than 95% of the cells were positive for each neurofilament protein, and 80% of the cells were positive for vimentin; less than 10% of the cells in S phase were positive with each of the five antineurofilament protein monoclonal antibodies (Mabs), but 20% of the vimentin-positive cells were in S phase. In the WERI-Rb1 line, 44 and greater than 96% of the cells were positive for the high-molecular-weight neurofilament subunit and high- and middle-molecular-weight neurofilament subunits proteins, respectively, but only 5% of the high-molecular-weight neurofilament positive cells were in S phase. In the U-251 MG line, 37 and 98% of the cells were positive for GFA protein and vimentin, respectively; only 3% of the GFA protein-positive cells, but 13% of the vimentin-positive cells, were in S phase. The results indicate that, when maintained in a matrix culture system, most cells in S phase in these lines lack markers of neural differentiation.
AuthorsN Ibayashi, M M Herman, J C Boyd, D D Bigner, H S Friedman, V P Collins, L A Donoso, L J Rubinstein
JournalLaboratory investigation; a journal of technical methods and pathology (Lab Invest) Vol. 61 Issue 3 Pg. 310-8 (Sep 1989) ISSN: 0023-6837 [Print] United States
PMID2504992 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Intermediate Filament Proteins
  • Neurofilament Proteins
  • Vimentin
  • Bromodeoxyuridine
Topics
  • Astrocytoma (metabolism)
  • Bromodeoxyuridine
  • Cell Cycle
  • Humans
  • Immunoenzyme Techniques
  • Intermediate Filament Proteins (analysis)
  • Interphase
  • Medulloblastoma (metabolism)
  • Neurofilament Proteins
  • Retinoblastoma (metabolism)
  • Tumor Cells, Cultured
  • Vimentin (analysis)

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