Mechanism Of MiR-203 Mediated Placental Development Via VEGFA Inhibition

PART ONE:Comparison of Gene Expression Profiles Between Normal and Preeclampsia Tissues via BioinformaticsVasculogenesis and extensive angiogenesis are indispensable for placental development,and it is well known that dysfunctions in these processes will lead to adverse pregnancy outcomes.Pre-eclampsia(PE)is one of the results of dysfunctional vascular development.The gene expression profiles of GSE9984 and GSE6573 were downloaded from the Gene Expression Omnibus database.Differentially expressed genes(DEGs)analysis was performed by GE02R.Gene Set Enrichment Analysis(GSEA)was conducted using the software GSEA v3.0.TargetScanHuman,miRTarBase,mirRDB are applied to analyze the common target genes of miR-203.Visualization of interaction networks of miR-203 with target genes was constructured by the Cytoscape.Total of 10 genes was selected after the comparison of the two gene profiles,including LNPEP,AKNA,SLC9A7,VEGFA,FUT9,STRBP,TMEM144,LPAR6,ERCC6L,TRAP1.Further comparing the 10 common genes with the target genes,it was found that interaction of miR-203 with VEGFA commonly existed in the PE development and placentation.PART TWO:MiR-203 Participates in Human Placental Angiogenesis by Inhibiting VEGFA and VEGFR2 ExpressionAngiogenesis during placentation is of great significance in maintaining normal pregnancy.However,the molecular mechanisms of this process are not clear.It has been reported that miR-203 plays a critical role in the development and progression of many tumors but not focused on the relationship between miR-203 and placental angiogenesis.The present study aims to illustrate the correlation between miR-203 and vascular endothelial growth factor(VEGFA)/vascular endothelial growth factor receptors 2(VEGFR2)in human placenta and human umbilical vein endothelial cells(HUVECs)obtained from 40 samples.Samples of human placenta were collected based on gestation age,which was divided into early preterm(n = 10),late preterm(n=12),and term(n=18).In this work,we demonstrated that the expression of miR-203 decreased significantly in the placenta according to the gestation age,in contrast,the expression of VEGFA and VEGFR2 increased accordingly.In vitro experiments revealed that overexpression of miR-203 not only suppressed the proliferation,migration,invasion,and tube formation of HUVECs but also down-regulated the expression of VEGFA and VEGFR2.Furthermore,inhibition of miR-203 expression showed equally apparent positive effects on HUVECs.In conclusion,our study suggests that miR-203 plays an important role in regulating placental angiogenesis through inhibiting the expression of VEGFA and VEGFR2,thus miR-203 may represent a potential therapeutic target for patients with abnormal formation of blood vessels in the placenta.PART THREE:MiR-203 Contributes to Pre-eclampsia by Inhibiting VEGFA ExpressionPre-eclampsia(PE)is a common but complex condition that can occur in pregnancy.It is estimated to affect 3%-8%of pregnancies worldwide.PE development is thought to be multifactorial and to involve the dysregulation of microRNA(miR)expression.However,the precise mechanisms of PE development remain unclear.The present study aimed to illustrate the association between miR-203 expression and PE development in samples of human placenta collected from mothers with(n=18)and without(n=20)PE.It was demonstrated that miR-203 expression was signifcantly increased in the PE placenta compared with the normal placenta samples,while the expression of vascular endothelial growth factor A(VEGFA)was decreased.In vitro experiments revealed that miR-203 overexpression signifcantly downregulated VEGFA expression and inhibited the proliferation,migration and inva sion ability of HTR 8/SVneo cells.Suppression of miR-203 expression alleviated these effects.A luciferase reporter assay confirmed the interaction of the 3’untranslated region of VEGFA with miR-203.Thus,miR-203 may have signifcant contribution to the development of PE by targeting VEGFA in the human placenta and may have potential as a biomarker or therapeutic target in the treatment of PE.