Role Of SASH1 In Trophoblast Cells And Its Mechanism In Preeclampsia

Objectives:To investigate the expression and localization of SASH1(SAM-and SH3-domain containing1)gene in placental tissues of normal pregnancy and preeclampsia.To investigate the role of SASH1 in regulating the function of HTR8/SVneo,including proliferation,migration,invasion and appoptosis;To explore the effect of SASH1 gene in regulating the function of migration and invasion of HTR8/SVneo cells cultured in hypoxic environment and to identify the molecular mechanisms underlined,to provide new ideas and therapeutic measures for the study of preeclampsia.Methods:(1)qPCR,western blotting and immunohistochemistry were used to detect the expression and localization of SASH1 in placenta,and compare the expression of SASH 1 in nomal placenta and placenta of preeclampsia.(2)Interfering the expression of SASH1 in HTR8/SVneo cells by Silencing and overexpression the SASH1 gene;(3)PCR was used to detect the interference efficiency;(4)The effects of SASH1 expression on the proliferation,migration,invasion and appoptosis of HTR81 SVneo cells were detected by CCK8 and Edu proliferation experiments,Transwell migration experiments,Matrigel invasion experiments and Annexin V-VGFP cell apoptosis Kit;(5)To further clarify the role of SASH1 in the regulation of trophoblast function and possible pathogenesis mechanism of preeclampsia,we established the hypoxia model of trophoblast cells by culture the HTR8/SVneo cells in hypoxia environment in vitro and observed the biological function of HTR8/SVneo cells.Results:(1)We can find the expression of SASH1 in the placentas of nomal pregnancy and preeclampsia,and SASH1 was mainly expressed in the cytoplasm and nucleus of trophoblast cells.Compared with the normal control group,the expression of SASH1 was higher in the placenta of preeclampsia;(2)The expression of SASH1 mRNA and protein in placenta of patients with preeclampsia was significantly higher than that in normal control group(P<0.05).(3)Knockdown of SASH1 gene could promote the proliferation,migration and invasion of HTR8/SVneo cells,and inhibit the rate of appoptosis;(4)Overexpression of SASH 1 gene could inhibit the proliferation,migration and invasion of HTR8/SVneo cells,and promote the appoptosis;(5)The treatment of hypoxia can reduce the expression of MMP2 mRNA and MMP9 mRNA in trophoblast cells;and the expression of MMP2 mRNA and MMP9 mRNA in trophoblast cells could relatively elevated after knckdown the SASH1 gene;(6)Knockdown of SASH1 gene,can effectively promote the HTR8/SVneo cells to repairt the function of migration and invasion induced by hypoxia injury;Conclusion:(1)SASH1 was expressed in the placentas of nomal pregnancy and preeclampsia;(2)SASH1 plays an important role in the process of trophoblastic proliferation,migration,invasion and appoptosis;(2)SASH1 may affects the migration and invasion of trophoblast cells during placental formation in preeclampsia by affecting the expression of MMP2/MMP9 of trophoblast cells;(3)knockdown of SASH1 gene can effectively improve HTR8/SVneo cell function injuried by the hypoxia injury,including migration and invasion,thereby improving the abnormalities of preeclampsia placental shallow bed,and providing new ideas and targets for preeclampsia Treatment.