The Role Of GnT-V In Regulating Trophoblast And Human Umbilical Vein Endothelial Cells Funtions:Possibl Erelevance To The Pathological Mechanism Of Preeclampsia

Objective: Preeclampsia(PE)is a human pregnancy-specific disease and a major contributor to maternal mortality worldwide.It is well known that PE is associated with shallow invasion of the trophoblast and insufficient remodeling of the uterine spiral artery.Accumulating evidence suggests that N-acetylglucosaminyltransferase V(GnT-V)is correlated with tumor invasion and metastasis.Our objective was to characterize GnT-V expression and function during the development of PE.Methods: The expression of GnT-V in placental tissue from the first trimester and trird trimeser was determined by immunohistochemistry.To investigate whether GnT-V regulates trophoblast/ endothelial cell function,we knockdown GnT-V expression through transfecting HTR8/SVneo,human umbilical vein endothelial cells(HUVECs)and villous explants with infected by the shRNA.We investigated invasion/migration of the HTR8/SVneo cells and the human villous explants,and migration and tube formation capacity of HUVECs by Transwell experiments.Cell proliferation and apoptosis were measured by MTT assay and flow cytometry(FCM),respectively.The activity of matrix metalloproteinase(MMP)2/9 and the expression of tissue inhibitors of metalloproteinases(TIMPs)1/2 were determined by gelatin zymography and Western blot.At last,the HTR8/SVneo,HUVECs and explants models were subjected to the H/R condition,in order to investigate the GnT-V expression and its effect on those cells in vitro.We use PD98059 blockage to study whether FAK-ERK signaling pathway was involved Gn T-V regulation cell functions of HTR8/SVneo cells and HUVECs.Results: In first trimester villi,GnT-V was specifically localized within the CTBs,the STBs and the TCs.In the maternal decidu,was expressed in DCs,ECs and some EVTs.The expression of GnT-V was elevated in PE placentas compared with the normal control placentas.GnT-V shRNA significantly enhanced the invasion and migration capability of HTR8/SVneo cells,the migration and tube formation potencial of HUVECs,and increased villous explant outgrowth.The enhanced effect of GnT-V shRNA on trophoblast cell invasion was associated with increased gelatinolytic activity of MMP2/9 and decreased expression of TIMP1/2.The expression of GnT-V was increased in H/R-exposed HTR8/SVneo cells and HUVECs.The knockdown of GnT-V and PD98059 treatment significantly enhanced cell migration/invasion of H/R-exposed HTR8/SVneo cells and promoted migration/tube formation capacity in H/R-exposed HUVECs.Conclsion: Our data suggest that oxidative stress induces the overexpression of GnT-V via the regulation of the focal adhesion kinase(FAK)–extracellular signalregulated kinase(ERK)signaling pathway,which,in turn,affects the function of trophoblast and endothelial cells,which then participates in the pathogenesis of PE.