The Regulation Of Trophoblast And Human Umbilical Vein Endothelial Cells Functions By ACK1 Which May Involved In The Pathological Mechanism Of Preeclampsia

Objective:Preeclampsia(PE)is a serious and common pregnancyspecific disease,it complicates 4% of pregnancy worldwide.Current research demonstrates that shallow invasion of trophoblast and insufficient remodeling of uterine spiral artery is due to the gestational hypertensive disease.In addition,the metalloproteinases plays a key role in the development.Current research suggests that ACK1 is associated with the tumor invasion and metastasis.Our study was to investigate ACK1 expression and function during the development of PE.Methods:We used immunohistochemistry to determine the expression of ACK1 in early and late trimester plcenta.We knockdown the expression of ACK1 through transfecting HTR8/Svneo and HUVECs with lentiviralvector-based short-hairpin RNA(shRNA).The transfection efficiency was investigated by qRT-PCR and western bloting.We used Transwell expriments to investigate the invasion/migration of the HTR8/Svneo cells,and the migration and tube formation capacity ofHUVECs also.We used CCK-8 assay and flow cytometry(FCM)to investigate the proliferation and apoptosis of cell lines,respectively.We used Western blot to determine the activity of Matrix metalloproteinase(MMP)2/9 activities and tissue inhibitor of metalloproteinase(TIMP)1/2.At last,the HTR8/Svneo cells and HUVECs were treated with H/R to detect the expression of ACK1 and its effect on those cells in vitro.In order to investigate whether PI3K-Akt signaling pathway was involved in the regulation of HUVECs and HTR8/Svneo function by ACK1,we used the PI3 K inhibitor LY294002.Results: The results showed that ACK1 was expressed in tropoblast cells and human endothelial cells in early pregnancy.Moreover,ACK1 expression in pre-eclampsia were much more lower than the control placentas.ACK1 shRNA not only decreased the invasion and migration capability of HTR8/Svneo cells significantly,but also the tube formation and migration capacity of HUVECs.ACK1 shRNA decrased these capacity of HTR8/Svneo cells and HUVECs through decreseing the activity MMP2/9 and increseing the expression of TIMP1/2.In H/R condition,the protein and mRNA expression of ACK1 was decreased not only in HTR8/Svneo cells but also HUVECs.ACK1 shRNA and LY294002 treatment significantly decreased cell migration/invasion of HTR8/Svneo cells,as well as migration/tube formation capacity in HUVECs.Conclsion: Our results showed that ACK1 enhanced the invasion andmigration of hunman trophoblast cell,as well as the angiogenesis of endothelial cells.The oxidative stress induces the decresed expression of ACK1 protein via Akt phosphorylation activation,which may be a possible pathological mechanism of PE.