Nestin expression in the cell lines derived from glioblastoma multiforme.
BACKGROUND
Nestin is a protein belonging to class VI of intermediate filaments that is produced in stem/progenitor cells in the mammalian CNS during development and is consecutively replaced by other intermediate filament proteins (neurofilaments, GFAP). Down-regulated nestin may be re-expressed in the adult organism under certain pathological conditions (brain injury, ischemia, inflammation, neoplastic transformation). Our work focused on a detailed study of the nestin cytoskeleton in cell lines derived from glioblastoma multiforme, because re-expression of nestin together with down-regulation of GFAP has been previously reported in this type of brain tumor.


METHODS
Two cell lines were derived from the tumor tissue of patients treated for glioblastoma multiforme. Nestin and other cytoskeletal proteins were visualized using imunocytochemical methods: indirect immunofluorescence and immunogold-labelling.


RESULTS
Using epifluorescence and confocal microscopy, we described the morphology of nestin-positive intermediate filaments in glioblastoma cells of both primary cultures and the derived cell lines, as well as the reorganization of nestin during mitosis. Our most important result came through transmission electron microscopy and provided clear evidence that nestin is present in the cell nucleus.


CONCLUSIONS
Detailed information concerning the pattern of the nestin cytoskeleton in glioblastoma cell lines and especially the demonstration of nestin in the nucleus represent an important background for further studies of nestin re-expression in relationship to tumor malignancy and invasive potential.
Veselska
Renata
R
Department of Biology, School of Medicine, Masaryk University, Brno, Czech Republic. rvesel@med.muni.cz
Kuglik
Petr
P
Cejpek
Pavel
P
Svachova
Hana
H
Neradil
Jakub
J
Loja
Tomas
T
Relichova
Jirina
J
D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000737:chemistry / Q000648:ultrastructure; D019084:Fluorescent Antibody Technique, Indirect; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005909:Glioblastoma / Q000737:chemistry* / Q000235:genetics / Q000473:pathology / Q000648:ultrastructure; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis*; D009419:Nerve Tissue Proteins / Q000032:analysis*; D064231:Nestin; D014746:Vimentin / Q000032:analysis
D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D014746:Vimentin