PMID- 12729802
OWN - NLM
STAT- MEDLINE
DCOM- 20030801
LR  - 20190707
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 286
IP  - 1
DP  - 2003 May 15
TI  - Noncatalytic domain of uPA stimulates human extravillous trophoblast migration by 
      using phospholipase C, phosphatidylinositol 3-kinase and mitogen-activated 
      protein kinase.
PG  - 138-51
AB  - The serine protease urokinase-type plasminogen activator (uPA) promotes matrix 
      degradation by many cell types, including the invasive extravillous trophoblast 
      (EVT) of the human placenta. The noncatalytic amino-terminal end of uPA binds to 
      uPA receptors (uPARs) expressed by these cells. A highly polarized expression of 
      uPAR-bound uPA at the migration front of EVT cells in situ suggests a functional 
      role of uPA:uPAR interaction in EVT cell migration. The present study examined 
      whether uPA stimulates EVT cell migration, independent of proteolytic function, 
      and investigated some of the signaling pathways involved. Using in 
      vitro-propagated EVT cells in Transwell migration assays, both uPA and its 
      noncatalytic amino-terminal fragment (ATF) were shown to stimulate migration 
      through multiporous polycarbonate (pore size 8 microm) membranes. A uPAR-blocking 
      antibody inhibited basal and ATF-stimulated migration. Migration was found to be 
      stimulated by hypoxic conditions, which upregulates uPAR expression; this 
      stimulation was abrogated with the uPAR-blocking antibody, indicating the role of 
      endogenous uPA in EVT cell migration. Spectrofluorometric measurement of 
      cytosolic calcium in cells treated with uPA and ATF demonstrated a rapid rise in 
      [Ca2+](i), which was prevented by pretreatment of cells with thapsigargin, 
      indicating a release from intracellular stores. Both basal and ATF-mediated 
      migratory responses were suppressed in the presence of selective pharmacological 
      inhibitors LY294002, U73122, and U0126, implicating the respective roles of 
      phosphatidinylinositol 3-kinase (PI 3-K), phospholipase C (PLC), and MEK1/2 in 
      basal and ATF-stimulated migratory capacity. Taken together, these results 
      demonstrate that uPA:uPAR interaction stimulates EVT cell migration, independent 
      of uPA enzymatic activity, using the mitogen-activated protein kinase pathway and 
      calcium signaling events including the participation of PI 3-K and PLC. These 
      findings are relevant to clinical conditions of aberrant trophoblast migration, 
      including spontaneous abortion, preeclampsia, and choriocarcinoma.
FAU - Liu, Jessica
AU  - Liu J
AD  - Department of Anatomy and Cell Biology, Medical Sciences Building, Faculty of 
      Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada 
      N6A 5C1.
FAU - Chakraborty, Chandan
AU  - Chakraborty C
FAU - Graham, Charles H
AU  - Graham CH
FAU - Barbin, Youssef P
AU  - Barbin YP
FAU - Dixon, S Jeffrey
AU  - Dixon SJ
FAU - Lala, Peeyush K
AU  - Lala PK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Peptide Fragments)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EC 3.1.4.- (Type C Phospholipases)
RN  - EC 3.4.21.- (Plasminogen Activators)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Calcium/metabolism
MH  - Cell Line
MH  - *Cell Movement
MH  - Culture Media, Serum-Free
MH  - Enzyme Inhibitors/metabolism
MH  - Humans
MH  - Mitogen-Activated Protein Kinases/*metabolism
MH  - Peptide Fragments/metabolism
MH  - Phosphatidylinositol 3-Kinases/*metabolism
MH  - Plasminogen Activators/chemistry/*metabolism
MH  - Protein Structure, Tertiary
MH  - Signal Transduction/physiology
MH  - Trophoblasts/cytology/*physiology
MH  - Type C Phospholipases/*metabolism
MH  - Urokinase-Type Plasminogen Activator/chemistry/*metabolism
EDAT- 2003/05/06 05:00
MHDA- 2003/08/02 05:00
CRDT- 2003/05/06 05:00
PHST- 2003/05/06 05:00 [pubmed]
PHST- 2003/08/02 05:00 [medline]
PHST- 2003/05/06 05:00 [entrez]
AID - S0014482703000892 [pii]
AID - 10.1016/s0014-4827(03)00089-2 [doi]
PST - ppublish
SO  - Exp Cell Res. 2003 May 15;286(1):138-51. doi: 10.1016/s0014-4827(03)00089-2.