New Pieces in the Puzzle of uPAR Role in Cell Migration Mechanisms

Cells. 2020 Nov 24;9(12):2531. doi: 10.3390/cells9122531.

Abstract

The urokinase (uPA) receptor (uPAR) plays a key role in cell migration. We previously showed that uPAR-negative HEK-293 cells efficiently migrate toward serum but, after uPAR ectopic expression, migrate only in a uPAR-dependent manner. In fact, migration of uPAR-transfected HEK-293 (uPAR-293) cells is impaired by anti-uPAR antibodies, without recovery of the uPAR-independent migration mechanisms formerly active. Prostate carcinoma PC3 cells, which express high endogenous uPAR levels, migrated only through a uPAR-dependent mechanism; in fact, the silencing of uPAR expression inhibited their migration. We hypothesize a crucial role of the uPAR glycosyl-phosphatidyl-inositol (GPI) tail, which promotes uPAR partitioning to lipid rafts, in uPAR-controlled cell migration. Here, we show that removal of the uPAR GPI-tail, or lipid rafts disruption by methyl-beta-cyclodextrin impairs migration of PC3 cells, incapable of uPAR-independent migration, whereas it restores uPAR-independent migration in uPAR-293 cells. We then show that, in PC3 cells, both uPAR signaling partners, β1 integrins and receptors for formylated peptides (FPRs), partly associate with lipid rafts. Inhibition of their interaction with uPAR impairs this association and impairs cell migration. Interestingly, blocking uPAR association with FPRs also impairs β1 integrin partitioning to lipid rafts, whereas blocking its association with β1 integrins has no effect on FPRs partitioning. On these bases, we propose that uPAR controls cell migration by connecting β1 integrins and FPRs and, through its GPI tail, by driving them into lipid rafts, thus promoting pro-migratory signals. uPAR-mediated partitioning of integrins to lipid rafts is strictly dependent on uPAR association with FPRs.

Keywords: FPR1; cell migration; fMLF receptors; uPAR; urokinase receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Movement / physiology*
  • HEK293 Cells
  • Humans
  • Integrin beta1 / metabolism
  • Membrane Microdomains / drug effects
  • Membrane Microdomains / metabolism
  • PC-3 Cells
  • Receptors, Urokinase Plasminogen Activator / metabolism*
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Urokinase-Type Plasminogen Activator / metabolism*
  • beta-Cyclodextrins / pharmacology

Substances

  • Integrin beta1
  • Receptors, Urokinase Plasminogen Activator
  • beta-Cyclodextrins
  • methyl-beta-cyclodextrin
  • Urokinase-Type Plasminogen Activator