Expression and function of the high affinity alphaIIbbeta3 integrin in murine melanoma cells.
In resting platelets integrin alphaIIbbeta3 is constitutively expressed in an inactive state and it does not recognize soluble proteins. Platelet activation results in a conformational change of the low-affinity alphaIIbbeta3 to a high-affinity state which then recognizes plasma fibrinogen. The ectopic expression of alphaIIbbeta3 integrin in rodent and human cells derived from solid tumors is well documented, although little is known about its affinity state in these tumor cells. In this study we analysed expression and function of high-affinity alphaIIbbeta3 in murine metastatic melanoma B16a cells by using a mAb that specifically recognizes high-affinity alphaIIbbeta3 (PAC-1). These tumor cells while in suspension bound PAC-1 and fibrinogen. Immunofluorescent studies of B16a cells indicated that high-affinity alphaIIbbeta3 is associated with the Golgi complex and the cell surface. Stimulation of B16a cells with a PKC-activator, 12(S)-HETE, induced translocation of the high-affinity integrin from an intracellular pool to the plasma membrane, which resulted in increased tumor cell adhesion to fibronectin. In addition to participating in 12(S)-HETE-stimulated adhesion of B16a cells, the high-affinity alphaIIbbeta3 integrin is also involved in tumor cell invasion through a reconstituted basement membrane. In conclusion, results from this study suggest that in non-megakaryocytic lineage B16a cells alphaIIbbeta3 is constitutively expressed in a high-affinity state, and that this conformation participates in tumor cell adhesion and invasion.
Timar
J
J
First Institute of Pathology and Experimental Cancer Research, Semmelweis University of Medicine, Budapest, Hungary. jtimar@korb1.sote.hu
Trikha
M
M
Szekeres
K
K
Bazaz
R
R
Honn
K
K
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D054641:Dual Specificity Phosphatase 2; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D008546:Melanoma, Experimental / Q000378:metabolism* / Q000473:pathology / Q000556:secondary; D051379:Mice; D009361:Neoplasm Invasiveness; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000096:biosynthesis* / Q000502:physiology; D054648:Protein Phosphatase 2; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism
D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen; D054648:Protein Phosphatase 2; C517032:DUSP2 protein, human; D054641:Dual Specificity Phosphatase 2; C517033:Dusp2 protein, mouse; D017027:Protein Tyrosine Phosphatases