A human colon carcinoma cell line exhibits adhesive interactions with P-selectin under fluid flow via a PSGL-1-independent mechanism.
It has been postulated that endothelial cell adhesion molecules involved in leukocyte recruitment play a role in metastasis. Using an in vitro flow model, we studied the adhesion of the human colon carcinoma cell line KM12-L4 to P-selectin, an inducible endothelial-expressed adhesion molecule involved in leukocyte recruitment. Recombinant forms of P-selectin and Chinese hamster ovary cells stably expressing P-selectin supported attachment and rolling of KM12-L4 cells at 1 to 2 dynes/cm2. The adhesive interactions to P-selectin were abolished by pretreatment of the KM12-L4 cells with neuraminidase but were unaltered by pretreatment of the KM12-L4 cells with O-sialoglycoprotein endopeptidase, an enzyme that cleaves mucin type glycoproteins such as P-selectin glycoprotein ligand-1 (PSGL-1). PSGL-1 is the only counter-receptor for P-selectin known to mediate myeloid cell adhesion to P-selectin under flow. Flow cytometric and Northern blot analyses revealed that KM12-L4 cells did not express PSGL-1 and monoclonal antibody PL1, a function-blocking monoclonal antibody to PSGL-1, had no inhibitory effect on KM12-L4 adhesion to P-selectin under flow. Compared with HL-60 cells, which express PSGL-1, the KM12-L4 cells exhibited a slightly lower rate of attachment to P-selectin and rolled at a significantly higher velocity. In summary, KM12-L4 human colon carcinoma cells interact with P-selectin, under flow, through a PSGL-1-independent adhesion pathway.
Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
D J
DJ
Goetz
H
H
Ding
W J
WJ
Atkinson
G
G
Vachino
R T
RT
Camphausen
D A
DA
Cumming
F W
FW
Luscinskas
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016466:CHO Cells; D002277:Carcinoma / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D003110:Colonic Neoplasms / Q000378:metabolism*; D006224:Cricetinae; D019040:E-Selectin / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000276:immunology / Q000494:pharmacology*; D008666:Metalloendopeptidases; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012212:Rheology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D019040:E-Selectin; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D011993:Recombinant Fusion Proteins; D008666:Metalloendopeptidases; C070738:O-sialoglycoprotein endopeptidase; D019158:N-Acetylneuraminic Acid