To further characterize the molecular mechanisms that govern carcinoma cell adhesion to stimulated endothelium, we studied the adhesion of a human colon carcinoma cell line, KM12-L4, to an E-selectin-IgG1 chimera and interleukin (IL)-1 beta-stimulated human umbilical vein endothelial cells (HUVEC) under in vitro fluid flow conditions. Between 0.6 and 1.8 dynes/cm2, KM12-L4 cells attach to and roll on IL-1 beta-stimulated HUVEC. The adhesion is E-selectin dependent and diminished upon pretreatment of the KM12-L4 cells with neuraminidase (neuraminidase sensitive). Between 0.7 and 1.8 dynes/cm2, surfaces coated with an E-selectin-IgG1 chimera support attachment and rolling of KM12-L4 cells. The adhesion to the E-selectin-IgG1 chimera is blocked by an antibody to the lectin domain of E-selectin and is neuraminidase sensitive. Rolling KM12-L4 cells exhibit variable velocity motion over both IL-1 beta-stimulated HUVEC and E-selectin-IgG1 chimera-coated surfaces. Our results provide the first direct evidence that sialylated moieties are involved in the adhesion of carcinoma cells to IL-1 beta-stimulated endothelium under flow conditions; E-selectin-IgG1 chimeras can support cell attachment and rolling under defined flow conditions; the topology of the endothelium is not the sole cause of variable velocity motion observed in cell rolling systems.
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