The Mechanism Of RND3 Negatively Regulating Trophoblast Cell Proliferation And Migration Invasion

Objective: In recent years,research has shown that the biological behavior of trophoblast plays an important role in the maintenance of pregnancy,especially the trophoblast cell apoptosis increased,poor invasion and inadequate capacity which cause placenta ischemia hypoxia by makingno enough oxygen to the embryo or the lack of nutrition factor.All of these eventually led to adverse pregnancy outcome.This study explored the differential expression and biological function of RND3 at the maternal-fetal interface by collecting samples of unexplained recurrent spontaneous abortion(u RSA)and establishing an in vitro trophoblast cell model.In addition,we explore the biological mechanism and molecular interaction affecting the differential expression of RND3,providing a basis for elucidating its important role at the maternal-fetal interface.Method: Real-time quantitative PCR,Western blot and immunohistochemistry were used to measure the expression levels of RND3 m RNA and proterin in villous tissues of u RSA and healthy controls(HCs).After overexpression or knockdown of RND3 in HTR8-S/Vneo trophoblastic cell line,CCK-8 assay,flow cytometry,Transwell assay and explant culture model were applied to explore the role of RND3 in regulation of trophoblast proliferation,apoptosis and migration.Contructing RND3 core promoter deletion fragment dual luciferase reporter plasmid and software prediction were used to find the important transcription factors that control RND3 transcription.Chromatin immunoprecipitation,dual luciferase reporter gene assay and site-directed mutagenesis were used to search for the transcriptional regulation of RND3 by transcription factors and to predict the transcriptional binding sites.The signal transduction pathway that RND3 regulates trophoblast cell function was studied by Western blot.Cell function experiment in vitro were further performed to verify the above regulatory mechanisms.Result: 1.The expression level of RND3 in trophoblast of u RSA villous tissues was signigicantely higer than that of HCs.2.Knockdown of RND3 promotes trophoblast cell proliferation,migration and inhibits trophoblast cell apoptosis;overexpression of RND3 inhibits trophoblast cell proliferation,migration and promotes trophoblast cell apoptosis.3.FOXD3 positively regulates RND3 transcription and its biological function in trophoblast cells.4.The expression level of FOXD3 was increased in cytotrophoblsts of u RSA villous tissues,and was positively correlated with the expression of RND3.5.RND3 regulates trophoblast migration and proliferation via the Rho A-ROCK1 signaling pathway and regulates apoptosis via ERK1/2signaling.Conclusions: RND3 inhibits trophoblast cell proliferation and migration via the Rho A-ROCK1 signaling pathway and promotes apoptosis via ERK1/2 signaling.The transcription factor FOXD3 positively regulates the transcription and biological functions of RND3.High levels of RND3 and FOXD3 are associated with u RSA in early pregnancy.