title: Focal adhesion kinase signaling is necessary for the Cyclosporin A-enhanced migration and invasion of human trophoblast cells.
abstract: OBJECTIVE
  Our previous studies have shown that Cyclosporin A (CsA) promotes human trophoblast invasion via mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway. E-cadherin and matrix metalloproteinases (MMPs) are important mediators in trophoblast migration and invasion. Here, we further investigate the role of focal adhesion kinase (FAK) signaling in the CsA-induced trophoblast migration and invasion and ERK activation.
  
  
  METHODS
  The migration and invasion of human primary trophoblasts and JEG-3 cells were measured by transwell migration and matrigel invasion assays. The activation of FAK, Src and ERK induced by CsA were examined by western blot. The colocalization of FAK and Src was detected by dual immunofluorescence assay. The regulation of E-cadherin expression and matrix metalloproteinases (MMPs) activity was evaluated by western blot and gelatin zymography, respectively.
  
  
  RESULTS
  CsA increased the phosphorylation of FAK and Src in human primary trophoblasts and JEG-3 cells. Meanwhile, the activated FAK and Src colocalized in the cytoplasm of JEG-3 cells. The FAK inhibitor Y15 or Src inhibitor PP2 could abrogate the phosphorylation of ERK, the enhanced migration and invasion and the activity of MMP2, 9 induced by CsA. In addition, these inhibitors also restored the expression of E-cadherin which is down-regulated by CsA. However, U0126, an inhibitor of ERK, had no significant effect on the CsA-induced activation of FAK and Src.
  
  
  CONCLUSIONS
  FAK-Src signaling, the upstream signaling cascade of ERK activation, plays an important role in the CsA-induced migration and invasion via down-regulating expression of E-cadherin and up-regulating activity of MMP2, 9 in trophoblast cells. These results may help provide a rationale to develop a novel therapeutic strategy for pregnancy disorders from insufficient trophoblast invasion.
authors: Laboratory for Reproductive Immunology, Hospital and Institute of Obstetrics and Gynecology, Fudan University Shanghai Medical College, 413 Zhaozhou Rd., Shanghai 200011, China.
authors: C-L
authors: CL
authors: Tang
authors: H-B
authors: HB
authors: Zhao
authors: M-Q
authors: MQ
authors: Li
authors: M-R
authors: MR
authors: Du
authors: Y-H
authors: YH
authors: Meng
authors: D-J
authors: DJ
authors: Li
mesh_terms: D015820:Cadherins / Q000032:analysis; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D016572:Cyclosporine / Q000494:pharmacology*; D003593:Cytoplasm / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000032:analysis / Q000502:physiology*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D011247:Pregnancy; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014327:Trophoblasts / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D019061:src-Family Kinases / Q000032:analysis / Q000378:metabolism
chemical_list: D015820:Cadherins; D007166:Immunosuppressive Agents; D016572:Cyclosporine; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9