LncAC079944.2 regulates endometrial receptivity via miR-149-5P/EFNA1 axis
Objective To screen and identify the lncRNA AC079944.2,which is closely associated with endometrial receptivity(ER),and further delved into its functional role and regulatory mechanism in ER.Methods Weighted gene co-expression network analysis(WGCNA)was performed on transcriptome data obtained from 71 endometrial tissue samples sourced from two datasets in the GEO database,leading to the identification of RNAs exhibiting significant correlation with ER.The consistency of bioinformatic-screened RNAs and their clinical expression was verified by analysis of endometrial samples from patients treated in the Department of Reproductive Medicine,the Fourth Affiliated Hospital of Jiangsu University from October 2021 to October 2022.Subsequently,the impact of lncAC079944.2 on Ishikawa cells'proliferation,migration,invasion was evaluated through cytofunctional experiments.The expression patterns of lncAC079944.2 and EFNA1,along with their interactions with screened miRNAs,were validated using dual luciferase reporter gene assays,qRT-PCR,and Western blotting analysis.Furthermore,their effects on cell function were confirmed.Results LncAC079944.2 was significantly upregulated in the middle secretory stage compared with the early secretory stage(P<0.001),and its expression in the middle secretory stage of recurrent implantation failure(RIF)patients was significantly lower than that of normal patients(P<0.001).Silencing the expression of lncAC079944.2 resulted in varying degrees of weakened proliferation(P=0.004),migration(P=0.001),and invasion(P<0.001)of Ishikawa cells.qRT-PCR revealed that silence of lncAC079944.2 led to a decrease in the expression of EFNA1(P=0.030),whereas the addition of miR-149-5p inhibitor resulted in the recovery of EFNA1 expression(P=0.034).Similarly,the addition of miR-149-5p inhibitor restored the weakened proliferation(P<0.001),migration(P=0.001)and invasion(P=0.008)abilities of Ishikawa cells,which had been compromised by silence of lncAC079944.2.In the embryo adhesion experiment,disruption of lncAC079944.2 expression significantly suppressed the adhesion between JAR cell spheres(simulated embryos)and Ishikawa cells(P<0.001).However,upon addition of a miR-149-5p inhibitor,there was a notable increase in the degree of adhesion between the embryos and Ishikawa cells(P<0.001).Conclusion LncAC079944.2 is associated with ER,and downregulating lncAC079944.2 can significantly decrease ER levels.lncAC079944.2 functions as a molecular sponge for miR-149-5p,modulating EFNA1 expression and impacting the proliferation,migration,invasion ability of Ishikawa cells,as well as influencing embryo adhesion to Ishikawa cells,thus affecting ER.LncAC079944.2 can regulate ER via the miR-149-5p/EFNA1 axis,which is expected to become a new indicator for the evaluation of endometrial receptivity,and provide experimental data for finding new strategies to improve ER.
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