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Enrichment of Collagen and Gelatin Degrading Activities in the Plasma Membranes of Human Cancer Cells¹

Department of Medicine and Research [S. Z., J. M. W., R. M. L., D. W.], Veterans Administration Medical Center, Northport, New York 11768 and Schools of Dental Medicine [N. S. R., L. M. G.] and Medicine [S. Z., B. L.], Health Science Center, State University of New York at Stony Brook, Stony Brook, New York 11794

Interactions between connective tissue substrates and proteinases localized to the surface of cancer cells are implicated in cancer invasion. In this report we have compared the enrichment of collagen and gelatin degrading activities and cysteine proteinase(s) in well-characterized (enzyme markers and electron microscopy) subcellular membrane fractions isolated from human small cell lung cancer lines (NCI-H69 and NCI-H82) and the RWP-1 pancreatic cancer line. With each cell line collagenolytic, gelatinolytic, and cysteine proteinase activities were enriched 5- to 128-fold in the plasma membrane fractions with differences noted between microvilli versus smooth membrane profiles. Incubation of tumor plasma membranes with methyl-³H-labeled collagen resulted in extensive degradation of the , ß, 1, and 2 chains, suggesting the combined action of metalloproteinases. Treatment of tumor plasma membranes with the chaotropic agent, 2 M KCl, did not diminish membrane collagenor gelatin-degrading activity, but extensively leached out the cysteine proteinase, suggesting that the latter enzyme is not an integral membrane protein. Enzyme inhibitors specific for metalloproteinases and cysteine proteinase were used to corroborate enzymatic classification. In conclusion, we have demonstrated variations in the localization of proteinases in the plasma membrane domains of different human cancer cells.

¹ Supported by Merit Review Research Funds from the Veterans Administration and USPHS Grant DE-03987 from the National Institute of Dental Research.

² To whom requests for reprints should be addressed, at Veterans Administration Medical Center, Northport, NY 11768.

Received 1/31/86. Revised 7/22/86. Revised 11/18/86. Accepted 12/11/86.

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