title: Co-expression of nestin and vimentin intermediate filaments in invasive human astrocytoma cells.
abstract: Intermediate filaments (IFs) are highly diverse intracytoplasmic proteins within the cytoskeleton which exhibit cell type specificity of expression. A growing body of evidence suggests that IFs may be involved as collaborators in complex cellular processes controlling astrocytoma cell morphology, adhesion and proliferation. As the co-expression of different IF subtypes has been linked to enhanced motility and invasion in a number of different cancer subtypes, we undertook the present study to examine the expression of vimentin and nestin in a panel of human astrocytoma cell lines whose tumorigenicity, invasiveness and cytoskeletal protein profiles are well known. Astrocytoma cells were examined for IF protein expression by immunofluorescence confocal and immunoelectron microscopy. The motility of all cell lines was determined by computerized time-lapse videomicroscopy. Invasive potential of astrocytoma cells was determined using Matrigel as a barrier to astrocytoma cell invasion in vitro. Vimentin was expressed by all astrocytoma cell lines. On the other hand, nestin was variably expressed among the different cell lines. The most motile and invasive astrocytoma cell line in our study was antisense GFAP-transfected U251 (asU251) astrocytoma cells which showed marked up-regulation of nestin expression compared to the U251 parental cell line and controls. The U87 astrocytoma cell line also demonstrated high nestin expression levels and was associated with an increased basal motility rate and a high degree of invasiveness through Matrigel. U343 astrocytoma cells did not express nestin, but had high levels of GFAP. It had the lowest motility rate and invasiveness of all the astrocytoma cell lines examined. Taken together, these data suggest that for the astrocytoma cell lines examined in this study, nestin and vimentin co-expression may serve as a marker for an astrocytoma cell type with enhanced motility and invasive potential. Further studies are required to determine the mechanism by which dual-IF protein expression alters other cytoskeletal or cell surface receptor protein components important in the process of astrocytoma invasion.
authors.affiliation: Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Division of Neurosurgery, The University of Toronto, Ont., Canada. rutka@sickkids.on.ca
authors.forename: J T
authors.initials: JT
authors.lastname: Rutka
authors.forename: S
authors.initials: S
authors.lastname: Ivanchuk
authors.forename: S
authors.initials: S
authors.lastname: Mondal
authors.forename: M
authors.initials: M
authors.lastname: Taylor
authors.forename: K
authors.initials: K
authors.lastname: Sakai
authors.forename: P
authors.initials: P
authors.lastname: Dirks
authors.forename: P
authors.initials: P
authors.lastname: Jun
authors.forename: S
authors.initials: S
authors.lastname: Jung
authors.forename: L E
authors.initials: LE
authors.lastname: Becker
authors.forename: C
authors.initials: C
authors.lastname: Ackerley
mesh_terms: D001254:Astrocytoma / Q000378:metabolism* / Q000473:pathology; D002465:Cell Movement / Q000502:physiology; D006801:Humans; D007381:Intermediate Filament Proteins / Q000096:biosynthesis*; D016253:Microscopy, Immunoelectron; D018715:Microscopy, Video; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D009419:Nerve Tissue Proteins; D064231:Nestin; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000096:biosynthesis*
chemical_list: D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; D014746:Vimentin