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.
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.
Ibayashi
N
N
Department of Pathology, University of Virginia School of Medicine, Charlottesville.
Herman
M M
MM
Boyd
J C
JC
Bigner
D D
DD
Friedman
H S
HS
Collins
V P
VP
Donoso
L A
LA
Rubinstein
L J
LJ
D001254:Astrocytoma / Q000378:metabolism*; D001973:Bromodeoxyuridine; D002453:Cell Cycle; D006801:Humans; D007124:Immunoenzyme Techniques; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007399:Interphase; D008527:Medulloblastoma / Q000378:metabolism*; D016900:Neurofilament Proteins; D012175:Retinoblastoma / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000032:analysis
D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins; D014746:Vimentin; D001973:Bromodeoxyuridine