HTRA4敲低对子痫前期滋养层细胞增殖和侵袭迁移影响OA

Objective To investigate the effect and mechanism of high-temperature requirement protein A4(HTRA4)on the proliferation,migration,and invasion of trophoblast cells in preeclampsia(PE).Methods Human trophoblast cell lines HTR-8/SVneo and JEG3 were used to establish an in vitro model of PE,and qRT-PCR and Western blot were used to measure the mRNA and protein expression levels of HTRA4.After HTRA4 was knocked down by shRNA,CCK-8 assay,flow cytometry,scratch assay,Transwell assay,and ELISA were used to measure the changes in cell viability,apoptosis,migration,invasion,and the epithelial-mesenchymal transition(EMT)markers E-Cadherin,N-Cadherin,and Vimentin.STRING database was used to pre-dict the interaction between HTRA4 and X-linked inhibitor of apoptosis protein(XIAP),and double immunofluorescent staining and co-immunoprecipitation were used to validate the direct binding between them.Co-transfection with sh-HTRA4 and sh-XIAP was performed to analyze the role of XIAP in HTRA4 regulation of trophoblast function.Results In the cell model of hypoxia-induced PE,there were significant increases in the protein expression level of HIF-1α and the mRNA and protein expression levels of HTRA4 G=4.49-38.04,P＜0.05).After knockdown of HTRA4,there was a significant increase in the viability of tropho-blast cells,a significant reduction in apoptosis rate,significant increases in migration and invasion abilities,a significant reduction in the level of E-Cadherin,and significant increases in the levels of N-Cadherin and Vimentin(F=7.60-506.90,P＜0.01).HTRA4 could directly bind to XIAP,and knockdown of HTRA4 could upregulate the expression of XIAP,while simultaneous si-lencing of XIAP could reverse the enhanced cell viability induced by HTRA4 knockdown(F=22.11,64.83,P＜0.05).Conclu-sion HTRA4 is involved in the pathogenesis of PE by negatively regulating the expression of XIAP and inhibiting the proliferation,migration,and invasion of trophoblast cells.