COMPUTERIZED ANALYSIS OF TUMOR-CELL INTERACTIONS WITH EXTRACELLULAR-MATRIX PROTEINS, PEPTIDES, AND ENDOTHELIAL-CELLS UNDER LAMINAR-FLOW

Arrest and formation of stable adhesive interactions between circulati ng cells and the endothelium or exposed subendothelial matrix are impo rtant processes in many biological situations. We have developed a hig hly sensitive hydrodynamic assay that utilizes a parallel-plate flow c hamber, video microscopy, and digital image processing to separate and measure the primary arrest and adhesion stabilization of flowing cell s. Our data indicate that primary cell contact triggers secondary adhe sion stabilization, and the secondary events are likely to be critical to metastasis formation. To study the relationship between tumor cell adhesion stabilization and organ-specific blood-borne metastasis, we investigated the adhesion stabilization of metastatic murine RAW117 la rge-cell lymphoma cells to the extracellular matrix proteins fibronect in and vitronectin, several Arg-Gly-Asp (RGD) containing peptides, and microvascular endothelial cells from the liver or lung. The highly li ver metastatic RAW117-H10 subline showed the fastest stabilization to fibronectin, vitronectin, and RGD peptides. Poorly metastatic RAW117-P cells had stabilization times 3-10 times longer than for RAW117-H10 c ells, while the lung- and liver-metastatic RAW117-L17 subline failed t o stabilize at all. The adhesion stabilization of the RAW117-H10 cells to the extracellular matrix proteins and RGD peptides was inhibited b y anti-beta(3) integrin monoclonal antibodies and RGD peptides. In con trast, the RAW117-L17 subline had the shortest stabilization time to u nstimulated microvascular endothelial cells of the lung and hepatic si nusoids, followed by RAW117-H10 cells and RAW117-P cells. Monoclonal a ntibodies against the beta(3) integrin subunit and RGD peptides did no t inhibit adhesion stabilization of RAW117-H10 cells to endothelial ce lls, suggesting that different metastatic variants of large-cell lymph oma cells use differing mechanisms to adhere to organ-specific endothe lial cells. (C) 1996 John Wiley & Sons, Inc.