Identification of a laminin-like substance on the surface of high-malignant murine fibrosarcoma cells.
High- and low-malignant murine fibrosarcoma cells were stained with anti-laminin antibodies using immunoperoxidase techniques and examined by electron microscopy. With the high-malignant cells, specific staining was observed along the cell surface. Use of normal rabbit serum in place of the rabbit anti-laminin or pretreatment of the anti-laminin with soluble laminin completely eliminated this staining. No immunoperoxidase staining was observed with the low-malignant cells. In additional studies, membrane fractions were prepared from the high- and low-malignant cells and used to immunize rabbits. The animals immunized with the membrane fractions from the high-malignant cells produced antibodies that reacted by enzyme-linked immunosorbent assay (ELISA) with murine laminin obtained from the EHS sarcoma. The animals immunized with membrane fractions from the low-malignant cells did not. These studies provide strong evidence that the high-malignant cells (but not the low) express on their cell surface a substance that is immunologically cross-reactive with laminin. In addition, the high-malignant cells (but not the low) secreted a material into the cell culture fluid that could be specifically immunoprecipitated with antilaminin antibodies. The immunoprecipitated material co-migrated with purified laminin when examined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis under reducing conditions. The existence of this substance associated with the surface of the high-malignant cells and its absence from that of the low-malignant cells may explain the previously noted difference between these cells in their ability to attach to type IV collagen. This difference may also contribute to the dissimilarity between these cells in their metastatic potential.
D000818:Animals; D000912:Antibodies, Neoplasm / Q000276:immunology; D000937:Antigen-Antibody Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005354:Fibrosarcoma / Q000032:analysis* / Q000648:ultrastructure; D007124:Immunoenzyme Techniques; D007797:Laminin / Q000032:analysis* / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D012513:Sarcoma, Experimental / Q000032:analysis; D000519:alpha-Galactosidase / Q000494:pharmacology
D000912:Antibodies, Neoplasm; D007797:Laminin; D008565:Membrane Proteins; D000519:alpha-Galactosidase