title: Cyclosporine A promotes in vitro migration of human first-trimester trophoblasts via MAPK/ERK1/2-mediated NF-κB and Ca2+/calcineurin/NFAT signaling.
abstract: BACKGROUND
  As a calcineurin inhibitor in T-cell activation, cyclosporine A (CsA) has provided the pharmacologic foundation for organ transplantation. We have previously demonstrated that CsA promotes trophoblast growth and invasion in vitro. Here, we further investigated the regulation of CsA on trophoblast migration and the intracellular signaling pathways involved.
  
  
  METHODS
  We evaluated the migration of human primary trophoblasts by using transwell migration assay. CsA-mediated induction of nuclear factor-kappa B (NF-κB) was evaluated by cotransfection with luciferase reporter constructs and luciferase activity assays.
  
  
  RESULTS
  Treatment with CsA-promoted migration of primary trophoblasts and the HTR8 cell line in a dose-dependent manner. CsA also increased NF-κB-transcriptional activity in trophoblasts in time- and dose-dependent manners. Pharmacologically inhibiting mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) 1/2 signaling with U0126 attenuated the CsA-induced cell migration and NF-κB activity in trophoblasts. Furthermore, pretreatment with PDTC, a specific NF-κB inhibitor, inhibited the CsA-induced migration of trophoblasts in dose-dependent manners. Although NFAT activation by ionomycin via calcineurin is accompanied by increased transactivation of NF-κB, pretreatment with the NFAT inhibitor did not affect NF-κB-transcriptional activity. Interestingly, ionomycin and CsA synergize to transactivate NF-κB. Ionomycin-inhibited basal migration of trophoblasts, and pretreatment with CsA reversed the ionomycin-inhibited trophoblast migration. However, the NFAT inhibitor increased basal migration, but not CsA-induced migration, of trophoblasts.
  
  
  CONCLUSIONS
  These observations indicate that both the MAPK/ERK/NF-κB pathway and Ca(2+)/calcineurin/NFAT pathways are involved in the CsA-promoted trophoblast migration. Our findings may help expand the clinical applications of this drug in trophoblast disorder.
authors: Laboratory for Reproductive Immunology, Hospital and Institute of Obstetrics and Gynecology, Fudan University Shanghai Medical College, 413# Zhaozhou Road, Shanghai 200011, China.
authors: S-C
authors: SC
authors: Wang
authors: Ch-L
authors: ChL
authors: Tang
authors: H-L
authors: HL
authors: Piao
authors: R
authors: R
authors: Zhu
authors: Ch
authors: Ch
authors: Sun
authors: Y
authors: Y
authors: Tao
authors: Q
authors: Q
authors: Fu
authors: D-J
authors: DJ
authors: Li
authors: M-R
authors: MR
authors: Du
mesh_terms: D019703:Calcineurin / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D016572:Cyclosporine / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005260:Female; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D050778:NFATC Transcription Factors / Q000502:physiology; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D015398:Signal Transduction / Q000187:drug effects; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology*
chemical_list: D004791:Enzyme Inhibitors; D016328:NF-kappa B; D050778:NFATC Transcription Factors; D016572:Cyclosporine; D048049:Extracellular Signal-Regulated MAP Kinases; D020928:Mitogen-Activated Protein Kinases; D019703:Calcineurin