Talin1/miR-1285-3p/PIPK1 Axis Regulating Endometrial Adhesion Via Influencing PI3K/AKT Signaling Pathway

Objective: To explore the role of Talin1 in endometrial adhesion and the underlying molecular mechanisms.Design: Analysis of Talin1/miR-1285-3p/PIPK1 axis influencing the endometrial adhesion and involved signaling pathway.Main Outcome Measure(s): In the first part,the expression of Talin1,FAK,Vinculin and integrin avβ3 in decidua and villi tissues were detected by real-time quantitative PCR(qPCR),western blotting(WB)and immunohistochemistry(IHC).In the second part,Talin1 and PIPK1 were silenced using RNA interference(RNAi)in endometrial cell lines and human trophoblast cell lines.The effect of Talin1 silencing(siTalin1)and PIPK1silencing(siPIPK1)on the expression of adhesion molecules(FAK/Vinculin/avβ3)were used qPCR and WB methods.Adhesion experiments were conducted to investigate the effects of siTalin1 and siPIPK1 on cell adhesion function.Flow cytometry was used to detect the effects of siTalin1 and siPIPK1 on cell cycle distribution and apoptosis level.The effect of siTalin1 and siPIPK1 on the expression of Cyclin D1/Cyclin E1/CDK4 was detected by qPCR and WB methods.In the third part,bioinformatics was used to predict the co-bindingmiRNA of Talin1 and PIPK1,and to search for the possible competitive binding sites of Talin1 and PIPK1.The luciferase reporter system confirmed that Talin1 and PIPK1 may competitively bind to the "seed sequence" of miR-1285-3p.The expression of PI3K/AKT signaling pathway related proteins were detected by WB after siTalin1 and siPIPK1.Result(s):1、Talin1 down-regulated in missed abortion tissues compared to normal abortion group2、Talin1 and PIPK1 knockdown inhibited expression of FAK,Vinculin and integrin-αvβ3 in Ishikawa and RL95-2 cells3、Talin1 and PIPK1 knockdown had no influence on the expression of FAK、Vinculin and integrin-αvβ3 in JAR and JEG-3 cells4、Talin1 and PIPK1 influenced cell adhesion of Ishikawa and RL95-2 cells5、Talin1 and PIPK1 knockdown suppressed cell proliferation and regulated cell cycle-related genes expression.6 、 Talin1 and PIPK1 competitively binded to the “seed squence” of miR-1285-3p7、Talin1/miR-1285-3p/PIPK1 axis regulated PI3K/AKT signaling pathwayConclusion(s):Our study has identified the function of Talin1/miR-1285-3p /PIPK1 signaling axis in the endometrial adhesion regulation from clinical tissue specimen,cellular biological function and potential molecular mechanism.