An invasion-related complex of cortactin, paxillin and PKCmu associates with invadopodia at sites of extracellular matrix degradation.
Invasive breast cancer cells have the ability to extend membrane protrusions, invadopodia, into the extracellular matrix (ECM). These structures are associated with sites of active matrix degradation. The amount of matrix degradation associated with the activity of these membrane protrusions has been shown to directly correlate with invasive potential. We demonstrate here that microinjection of polyclonal anti-cortactin antibodies blocks matrix degradation at invadopodia supporting the hypothesis that cortactin has a direct role in invasive behavior. MDA-MB-231, invasive breast cancer cells were sheared from the surface of a gelatin matrix to isolate invadopodia. Cortactin, paxillin and protein kinase C (PKC) mu, a serine kinase, were co-immunoprecipitated as a complex from invadopodia-enriched membranes. We confirmed the subcellular distribution of these proteins by immunolocalization and Western blotting. We also determined that, in contrast to its presence in invasive cells, this complex of proteins was not detected in lysates from non-invasive cells that do not form invadopodia. Taken together, these data suggest that the formation of this cortactin-containing complex correlates with cellular invasiveness. We hypothesize that this complex of molecules has a role in the formation and function of invadopodia during cellular invasion.
Bowden
E T
ET
Department of Cell Biology, Lombardi Cancer Center, Georgetown University Medical School, Washington, DC 20007, USA.
Barth
M
M
Thomas
D
D
Glazer
R I
RI
Mueller
S C
SC
D000199:Actins / Q000378:metabolism; D001943:Breast Neoplasms / Q000473:pathology* / Q000648:ultrastructure; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002462:Cell Membrane / Q000473:pathology / Q000648:ultrastructure*; D051356:Cortactin; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D005109:Extracellular Matrix / Q000502:physiology*; D005260:Female; D005780:Gelatin; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology; D008840:Microfilament Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008954:Models, Biological; D009361:Neoplasm Invasiveness; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000199:Actins; C495868:CTTN protein, human; D015815:Cell Adhesion Molecules; D051356:Cortactin; D003598:Cytoskeletal Proteins; D019012:Integrin beta1; D008840:Microfilament Proteins; C496061:PXN protein, human; D051419:Paxillin; D010750:Phosphoproteins; D005780:Gelatin; C089212:protein kinase D; D011493:Protein Kinase C