Immunochemical localization of heparanase in mouse and human melanomas.
Heparanase, an endo-beta-D-glucuronidase, has been associated with melanoma metastasis. Polyclonal antibodies directed against the murine N-terminal heparanase peptide detected a Mr approximately 97,000 protein on SDS-PAGE of mouse melanoma and human melanoma cell lysates. In an indirect immunocytochemical study, human A375-SM and mouse B 16-BL6 melanoma cells were stained with the anti-heparanase antibodies. Heparanase antigen was localized in the cytoplasm of permeabilized melanoma cells as well as at the cell surface of unpermeabilized cells. Immunohistochemical staining of frozen sections from syngeneic mouse lungs containing micrometastases of B16-BL6 melanoma demonstrated heparanase localized in metastatic melanoma cells. Similar studies using frozen sections of malignant melanomas resected from patients indicated that heparanase is localized in invading melanoma cells. Our studies suggest that (a) the N-terminus of the heparanase molecule in mouse and human is antigenically related; (b) heparanase antigens are localized at the cell surface and in the cytoplasm of metastatic human and mouse melanoma cells; and (c) heparanase antigens are enriched in invasive and metastatic murine and human melanomas in vivo.
Jin
L
L
Department of Tumor Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Nakajima
M
M
Nicolson
G L
GL
D000818:Animals; D000906:Antibodies / Q000032:analysis; D001345:Autoradiography; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000276:immunology; D003593:Cytoplasm / Q000201:enzymology / Q000276:immunology; D005966:Glucuronidase; D006026:Glycoside Hydrolases / Q000138:chemical synthesis / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007114:Immunization; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000201:enzymology / Q000276:immunology / Q000556:secondary; D008545:Melanoma / Q000201:enzymology* / Q000276:immunology; D008546:Melanoma, Experimental / Q000201:enzymology* / Q000276:immunology; D051379:Mice; D008819:Mice, Nude; D011233:Precipitin Tests; D011817:Rabbits; D014407:Tumor Cells, Cultured / Q000201:enzymology / Q000276:immunology; D000099175:Heparanase
D000906:Antibodies; D005966:Glucuronidase; D006026:Glycoside Hydrolases; D000099175:Heparanase