H19 Regulated Proliferation,Invasion,and Autophagy In The Placenta By Targeting MiR-18a-5p

Objective:To determine the role of long non-coding RNA H19 in fetal growth restriction?FGR?and to investigate the regulation of LncRNAH19on trophoblast proliferation,invasion and autophagy.Methods:Forty placental tissues were collected,half is normal,half is FGR.Real-time fluorescent quantitative PCR was used to detect the expression of LncRNA H19 in FGR pregnant women and normal pregnant women.Human chorionic trophoblast cells HTR8/Svneo and human choriocarcinoma cells JEG3 were cultured.Cells biological behavior and autophagy were tested after downexpression of H19.Target prediction revealed that LncRNA H19 may bind to miR-18a-5p and inhibit its activity,MiR-18a-5p expression was detected in placental tissues and its effect on cell biological behavior was tested in cells.Results:In the placental tissues with fetal growth restriction,the expression of LncRNA H19 was lower than that of the control group.The expression of H19 in HTR8 and JEG3 cells was decreased by siRNA interference technique,5-Ethynyl-2^'-deoxyuridine-based proliferation assay and CCK8 were used to test cell proliferation,and cell invasion ability was detected by Transwell assay.Western blotting was used to detect the expression of autophagy pathway related proteins.Analyses utilizing the online target sequence prediction database suggested that miR-18a-5p may have potential binding sites to H19,the expression of miR-18a-5p was higher in FGR than the control group,and the expression of miR-18a-5p was negatively correlate with the expression of LncRNAH19;Dual-luciferase reporter gene assay shown that H19 can directly bind with miR-18a-5p.The expression of LncRNA H19 was decreased after transfection of miR-18a-5p mimics,however,The expression of LncRNA H19 was increased after transfection of miR-18a-5p inhibitor.The expression of miR-18a-5p was increased after transfected with siH19.In the rescue experiment,siH19 and miR-18a-5p inhibitor were co-transfected to detect cell proliferation,invasion,autophagy and effect on IRF2.After co-transfected,the effect of H19 in proliferation,invasion,autophagy was diminished.Conclusion:LncRNA H19 expression is decreased and the expression of miR-18a-5p is increased in fetal growth-restricted placental tissues.Silencing H19 can inhibit cells proliferation,invasion and promote autophagy in fetal growth restriction by targeting miR-18a-5p.We hypothesized that dysregulation of H19/miR-18a-5p mediated regulatory pathway may contribute to the underlying mechanism of FGR.