Extracellular signal-regulated kinase functions in the urokinase receptor-dependent pathway by which neutralization of low density lipoprotein receptor-related protein promotes fibrosarcoma cell migration and matrigel invasion.
The low density lipoprotein receptor-related protein (LRP) has been reported to regulate cellular migration. In this study, an antisense RNA expression strategy was used to reduce LRP to undetectable levels in HT 1080 fibrosarcoma cells. The LRP-deficient cells demonstrated increased levels of cell-surface uPAR, higher levels of uPA in conditioned medium, increased migration on vitronectin-coated surfaces, and increased invasion of Matrigel. LRP-deficient cells also demonstrated increased levels of phosphorylated extracellular signal-regulated kinase (ERK) in the absence of exogenous stimulants. Antibodies which block binding of endogenously produced uPA to uPAR reduced ERK phosphorylation and migration of LRP-deficient cells to the levels observed with control cells. Inhibitors of ERK activation, including PD098059 and dominant-negative MEK1, also decreased the migration of LRP-deficient but not control cells. By contrast, constitutively active MEK1 stimulated the migration of control but not LRP-deficient cells. Although Matrigel invasion by LRP-deficient cells was inhibited by the proteinase inhibitor, aprotinin, PD098059 in combination with aprotinin was necessary for an optimal effect. Expression of the VLDL receptor in LRP-deficient cells reversed the changes in cellular migration and invasion. These studies demonstrate that binding of endogenously produced uPA to uPAR may serve as a major determinant of basal levels of activated ERK and, by this mechanism, regulate cellular migration and invasion. By regulating the uPA/uPAR system, LRP may also regulate ERK activation, cellular migration, and invasion.
Webb
D J
DJ
Departments of Pathology, Biochemistry and Molecular Genetics, University of Virginia Health Sciences Center, Charlottesville, VA 22908, USA.
Nguyen
D H
DH
Gonias
S L
SL
D007611:Aprotinin / Q000494:pharmacology; D019898:Autocrine Communication; D002465:Cell Movement / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D004789:Enzyme Activation / Q000187:drug effects; D005354:Fibrosarcoma / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000378:metabolism; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D048369:MAP Kinase Kinase 1; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D009361:Neoplasm Invasiveness; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases; D011509:Proteoglycans / Q000378:metabolism; D016372:RNA, Antisense / Q000235:genetics / Q000502:physiology; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D011973:Receptors, LDL / Q000235:genetics / Q000502:physiology; D055293:Receptors, Urokinase Plasminogen Activator; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors / Q000378:metabolism; D019096:Vitronectin / Q000378:metabolism
D017077:Culture Media, Conditioned; D004338:Drug Combinations; D005419:Flavonoids; D007797:Laminin; D026503:Low Density Lipoprotein Receptor-Related Protein-1; C524898:PLAUR protein, human; D011509:Proteoglycans; D016372:RNA, Antisense; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D011973:Receptors, LDL; D055293:Receptors, Urokinase Plasminogen Activator; C036053:VLDL receptor; D019096:Vitronectin; C059485:matrigel; D003094:Collagen; D007611:Aprotinin; D017346:Protein Serine-Threonine Kinases; D020928:Mitogen-Activated Protein Kinases; D048369:MAP Kinase Kinase 1; C482125:MAP2K1 protein, human; D020929:Mitogen-Activated Protein Kinase Kinases; D014568:Urokinase-Type Plasminogen Activator; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one