title: Neurotrophin stimulation of human melanoma cell invasion: selected enhancement of heparanase activity and heparanase degradation of specific heparan sulfate subpopulations.
abstract: Heparanase is an endo-beta-D-glucuronidase, the enzymatic targets of which are the glycosaminoglycan chains of heparan sulfate proteoglycans. Elevated levels of heparanase are associated with the metastatic potential of melanoma cells. Treatment of murine and human melanoma cells with the prototypic neurotrophin nerve growth factor (NGF) increases the production of heparanase by melanoma cells. We reported previously that physiological concentrations of NGF increased in vitro Matrigel invasion of early-passage human brain-metastatic 70W melanoma cells but not melanoma cells metastatic to other sites or nonmetastatic melanoma cells. Here we found that treatment of 70W melanoma cells with neurotrophin NT-3 increased Matrigel invasion, whereas treatment with neurotrophins other than NGF or NT-3 did not influence invasion. Mutants of NGF that do not bind to the neurotrophin receptor p75NTR or other nonneuronal growth factors were not able to enhance the invasion of 70W melanoma cells. When 70W cells were exposed to antisense oligonucleotides directed against p75NTR mRNA, there was a reduction in NGF and NT-3 binding, and the neurotrophins failed to enhance Matrigel invasion. To study the properties of heparanase in NT-regulated malignant melanoma invasive processes, we developed a sensitive heparanase assay consisting of purified [35S]heparan sulfate subpopulations separated by agarose gel electrophoresis. Incubation of 70W cells with NGF or NT-3, but not brain-derived NT factor, NT-4/5, or mutant NGF, resulted in increased release of heparanase activity that was capable of degrading a subpopulation of heparan sulfate molecules.
authors.affiliation: Department of Tumor Biology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, USA.
authors.forename: D
authors.initials: D
authors.lastname: Marchetti
authors.forename: D J
authors.initials: DJ
authors.lastname: McQuillan
authors.forename: W C
authors.initials: WC
authors.lastname: Spohn
authors.forename: D D
authors.initials: DD
authors.lastname: Carson
authors.forename: G L
authors.initials: GL
authors.lastname: Nicolson
mesh_terms: D001483:Base Sequence; D001932:Brain Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology / Q000556:secondary; D019208:Brain-Derived Neurotrophic Factor; D003094:Collagen / Q000378:metabolism; D004338:Drug Combinations; D004587:Electrophoresis, Agar Gel; D005966:Glucuronidase; D006025:Glycosaminoglycans / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000378:metabolism*; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000378:metabolism; D008545:Melanoma / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology / Q000556:secondary; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009361:Neoplasm Invasiveness; D009414:Nerve Growth Factors / Q000378:metabolism / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000378:metabolism / Q000494:pharmacology; D020933:Neurotrophin 3; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor / Q000187:drug effects / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D000099175:Heparanase
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D003094:Collagen; D004338:Drug Combinations; D005966:Glucuronidase; D006025:Glycosaminoglycans; D006026:Glycoside Hydrolases; D006497:Heparan Sulfate; D007797:Laminin; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D020933:Neurotrophin 3; D016376:Oligonucleotides, Antisense; D011509:Proteoglycans; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor; D000099175:Heparanase; C059485:matrigel; C071951:neurotrophin 5; C073922:neurotrophin 4