Altered production of laminin and nidogen by high and low metastatic variants of murine melanoma cells.
An altered ability to interact with and degrade extracellular matrix molecules is a common feature of the malignant phenotype. Although changes in the expression of matrix proteins in metastases in vivo are relatively well documented, little is known about the changes in matrix production by malignant cells in culture. Here we have examined the synthesis of the basement membrane components laminin and nidogen (entactin) by low and high metastatic variants of the K-1735 murine melanoma cells. Protein deposition was examined by western blotting as well as immunofluorescence; protein synthesis was examined by immunoprecipitation with specific antibodies. Gene expression was also evaluated by measuring steady-state mRNA levels using cDNA probes on northern and dot-blots. Laminin gamma 1 levels appeared to be similar in both high and low metastatic lines; however, the high metastatic lines had reduced levels of the laminin beta 1 chain. On the contrary, nidogen expression was observed only in the high metastatic lines. Traces of a laminin alpha chain were present only in immunoprecipitates of the low metastatic cells and could not be detected in the high metastatic cells. Both high and low metastatic cells deposited an extracellular matrix of basement membrane components, with laminin deposition decreased in high metastatic cells. Modified expression, production, and deposition of basement membrane components in high metastatic melanoma cells could be involved in their altered interactions with the extracellular matrix.
Valente
P
P
Department of Experimental Oncology, Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Noonan
D M
DM
Ogle
R C
RC
Albini
A
A
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D005109:Extracellular Matrix / Q000378:metabolism; D007163:Immunosorbent Techniques; D007797:Laminin / Q000096:biosynthesis* / Q000276:immunology; D008546:Melanoma, Experimental / Q000378:metabolism* / Q000556:secondary*; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000276:immunology; D051379:Mice; D012878:Skin Neoplasms / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D007797:Laminin; D008562:Membrane Glycoproteins; C029967:nidogen