Overexpression of urokinase receptor increases matrix invasion without altering cell migration in a human osteosarcoma cell line.
Proteolysis triggered by receptor-bound urokinase-type plasminogen activator (uPA) involves a cascade of species-specific molecular interactions. To study the role of the uPA receptor (uPAR) in such interactions, a human osteosarcoma cell line (HOS), which normally expresses low levels of uPAR, was transfected with human uPAR complementary DNA. One of several stably transformed clonal cells lines, designated 2A2, was characterized and compared to the parental HOS, revealing the following: (a) stable incorporation of uPAR complementary DNA into the genome demonstrated by Southern blot analysis; (b) a 10-fold increase in steady state mRNA levels of uPAR assessed by Northern blot analysis; (c) a 2-fold increase in the surface expression of glycosylphosphatidylinositol anchored uPAR protein determined by enzyme-linked immunosorbent assay and by the specific binding of radiolabeled single chain uPA; (d) a 2-fold increase in internalization and degradation of radiolabeled uPA/PAI-1 complexes; and (e) a 2-fold increase in receptor-bound uPA-mediated plasmin generation measured by the cleavage of a chromogenic substrate and degradation of 125I-labeled laminin. The involvement of uPAR in cellular processes was determined by comparing 2A2 and HOS cells in in vitro migration and invasion assays. The migration of 2A2 cells were slower on fibronectin-coated surfaces in a linear under-agarose assay, but both cell lines migrated at the same rate on uncoated polycarbonate filters in Boyden chamber assays. In the invasion experiments, 4 times more 2A2 than HOS cells penetrated through the barrier of reconstituted basement membrane Matrigel. These data suggest that uPAR does not potentiate random cell migration but facilitates matrix degradation and subsequent cell invasion.
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
K
K
Karikó
A
A
Kuo
D
D
Boyd
S S
SS
Okada
D B
DB
Cines
E S
ES
Barnathan
D001483:Base Sequence; D002465:Cell Movement / Q000502:physiology; D004247:DNA / Q000235:genetics; D005109:Extracellular Matrix / Q000502:physiology*; D005341:Fibrinolysin / Q000096:biosynthesis; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness; D012516:Osteosarcoma / Q000235:genetics* / Q000473:pathology* / Q000648:ultrastructure; D014176:Protein Biosynthesis / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000502:physiology; D055293:Receptors, Urokinase Plasminogen Activator; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D004247:DNA; D005341:Fibrinolysin