The effect of hsa-circRNA-103716 targeting miR-574-5p on endometrial receptivity in patients with repeated implantation failure
Objective To detect the expression of hsa-circRNA-103716 and miR-574-5p in human endometrium,and explore the role of both in the endometrial receptivity of patients with repeated implantation failures.Methods Detect the expression of hsa-circRNA-103716 and miR-574-5p in endometrial tissue and serum of 70 patients with recurrent implantation failure of frozen thawed embryo transfer(RIF group)and 70 patients with successful pregnancy after embryo transfer(control group)at planting window period by qRT-PCR,and to analyze their correlation with endometrial receptive pathological factors.Human primary endometrial epithelial cells(EECs)were divided into NC group,hsa-circRNA-103716 negative control(si-control)group,hsa-circRNA-103716 knockdown(si-circRNA)group,miR-con group,miR-574-5p overexpression(mimic)group,hsa-circRNA-103716+pcDNA group,and hsa-circRNA-103716+pcDNA-miR-574-5p group,CCK-8 method,EdU method,and cell scratch assay were used to detect the proliferation and migration ability of cells in each group.Western blot-ting was used to detect the expression of endometrial receptivity related protein.Verify the targeting relationship between hsa-circRNA-103716 and miR-574-5p by using the dual luciferase reporter gene method.Results The expression of hsa-circRNA-103716 in the endometrium and serum of the RIF group during the implantation window period was higher than that of the control group,while the expression of miR-574-5p was significantly lower than that of the control group,with a significant difference(all P＜0.001).The expression of hsa-circRNA-103716 was not related to age,body mass index,history of miscarriage,and infertility years(P＞0.05),but significantly correlated with endometrial thickness,morphology,and volume(P＜0.05).Compared with si-control group,EECs cell activity,EdU positive rate,cell mobility,LIF,VEGF,OPN,HOXA10,glycodelin-A and PRs protein expression levels of si-circ RNA group were significantly increased,but the expressions of TNF-α,ERs and IL-16 were significantly decreased,and the differences were statistically significant(all P＜0.05).After co-transfection with hsa-circRNA-103716 and miR-574-5p,EECs cell activity,EdU positive rate,cell mobility,LIF,VEGF,OPN,HOXA10,glycodelin-A and PRs protein expression levels were significantly increased,but the expressions of TNF-α,ERs and IL-16 were significantly decreased,and the differences were statistically significant(all P＜0.05).After co-transfecti-on of miR-574-5p with WT-hsa-cicRNA-03716 into EEC cells,the luciferase activity in the miR-574-5p mimic group was lower than that in the miR-con group.The difference was significant(P＜0.05).Conclusion hsa-circRNA-103716 is highly expressed in the endometrium of patients with repeated implantation failure,while miR-574-5p is low in expression,hsa-circ-RNA-103716 can reduce endometrial receptivity in patients with recurrent implantation failure by targeting miR-574-5p.
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