Purification of motility factor (GMF) from human malignant glioma cells and its biological significance in tumor invasion.
Two molecular species of glioma-derived motility factor (GMF), GMF-I and GMF-II, have been purified to homogeneity from the serum-free conditioned medium of a highly invasive human glioma cell line, T98G, by gelatin affinity chromatography and heparin affinity-, DEAE-, hydroxyapatite-, gel permeation- and sulfopropyl high performance liquid chromatography. GMF-I and GMF-II showed a single protein band on non-reducing SDS-polyacrylamide gel electrophoresis with a molecular mass of 145 kDa and 165 kDa, respectively. The physico-chemical characteristics of these two GMFs are similar with respect to amino acid composition. GMF-I and GMF-II both stimulated the migration of T98G cells in a concentration-dependent manner, and the activity of GMF-I was about 5 times as strong as that of GMF-II. Checkerboard analysis demonstrated that the GMFs had not only a chemotactic effect but also a chemokinetic effect on T98G cells. C6 glioma cells and T98G cells, both of which showed high invasiveness in an in vitro invasion assay with reconstituted basement membrane, Matrigel, migrated to the GMFs with great intensity, while A172 and 9L glioma cells and normal glial cells, all of which weakly infiltrated the Matrigel barrier, migrated to the GMFs with much less intensity. These results indicate that migratory response of glioma cells to the GMFs correlates well with invasiveness, suggesting an important role(s) of the GMFs in the process of glioma cell invasion.
Department of Neurosurgery, Osaka University Medical School, Japan.
T
T
Ohnishi
N
N
Arita
T
T
Hayakawa
K
K
Kawahara
K
K
Kato
A
A
Kakinuma
D000596:Amino Acids / Q000032:analysis; D002633:Chemotaxis / Q000187:drug effects*; D002845:Chromatography; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D020904:Glia Maturation Factor; D005910:Glioma / Q000473:pathology*; D006801:Humans; D006882:Hydroxyapatites; D008970:Molecular Weight; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000302:isolation & purification* / Q000494:pharmacology*; D012044:Regression Analysis; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D020904:Glia Maturation Factor; D006882:Hydroxyapatites; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D017886:Durapatite