Confocal fluorescence microscopy of urokinase plasminogen activator receptor and cathepsin D in human MDA-MB-231 breast cancer cells migrating in reconstituted basement membrane.
Using confocal fluorescence microscopy with a monoclonal antibody, we have localized the receptor for urokinase plasminogen activator (uPAR) in MDA-MB-231 human breast cancer cells migrating into a reconstituted basement membrane. Patchy and polarized uPAR immunoreactivity was found at the cell membrane, and strong staining was found both in the ruffled border or leading edge of the cells and at pseudopodia penetrating into the membrane. Intracellular uPAR staining was localized in the paranuclear region and in rounded granule-like structures; some of these were identified as lysosomes by double staining for uPAR and the lysosomal enzyme cathepsin D. Urokinase plasminogen activator (uPA) activity has previously been shown to play a role in migration of cells into basement membranes, and it has been proposed that uPAR also is involved in this process. uPA is known to be internalized and degraded after complex formation with the inhibitor PAI-1. Lysosomal uPAR immunoreactivity may result from concomitant internalization of the receptor.
Bastholm
L
L
Institute of Pathological Anatomy, University of Copenhagen, Denmark.
Nielsen
M H
MH
De Mey
J
J
Danø
K
K
Brünner
N
N
Høyer-Hansen
G
G
Rønne
E
E
Elling
F
F
D000911:Antibodies, Monoclonal / Q000276:immunology; D001485:Basement Membrane / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism* / Q000648:ultrastructure; D002402:Cathepsin D / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D008247:Lysosomes / Q000201:enzymology; D008856:Microscopy, Fluorescence; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D013194:Staining and Labeling; D016707:Tissue Fixation; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D002402:Cathepsin D