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Laminin, a Noncollagenous Component of Epithelial Basement Membranes Synthesized by a Rat Yolk Sac Tumor¹

The University Institute of Pathological Anatomy, Frederik V's 11, DK-2100 Copenhagen Ø, Denmark [U. W., R. A.], and La Jolla Cancer Research Foundation, La Jolla, California [E. R.]

Laminin, a glycoprotein antigenically similar or identical to a component of epithelial basement membranes, was identified as a major component of the abundant extracellular matrix synthesized by an experimentally induced rat yolk sac tumor. Immunocytochemical staining revealed laminin in cultured tumor cells as well as in their extracellular matrix. The presence of soluble laminin in the culture media of the tumor cells was demonstrated using metabolic labeling followed by identification by immunoprecipitation and sodium dodecyl sulfate:polyacrylamide gel electrophoresis. This revealed two polypeptides with molecular weights of approximately 200,000 and 400,000. These comigrated with the polypeptides of mouse laminin isolated previously.

The yolk sac tumor tissue grown in vivo contained laminin in the tumor cells and in the extracellular material as evidenced by immunofluorescence and immunoperoxidase staining. Immunization with the tumor matrix resulted in an antiserum that contained antilaminin and antifibronectin and was made specific for laminin by absorption with fibronectin. This antiserum precipitated laminin polypeptides from culture medium of yolk sac tumor cells and stained basement membranes in rat tissues in a manner indistinguishable from antilaminin. The presence of laminin in rat yolk sac cells, the presumed origin of our yolk sac tumor, was studied in some detail. Laminin was found to be present in normal cells of the visceral as well as the parietal yolk sac layer and in their basement membranes suggesting, but not proving, that both types of cells have ability to synthesize laminin.

Production of laminin and the presence of laminin-containing basement membrane material may be important for the biological behavior of the yolk sac tumor. This tumor will also be a useful source of laminin for chemical and biological characterization of this basement membrane protein.

¹ Supported by a grant from the Danish Cancer Society and by Grants CA 27455 and CA 28101 from the National Cancer Institute, Department of Health and Human Services.

² To whom requests for reprints should be addressed.

Received 9/ 8/80. Accepted 1/ 9/81.