Abstract
BackgroundVarious circular RNAs (circRNAs) are dysregulated in the placenta of fetal growth restriction (FGR) fetuses, but their roles and regulatory mechanisms have not been fully elucidated. Herein, we aimed to elucidate the role of hsa_circ_0081343 in regulating the migration, invasion, and apoptosis of human extravillous trophoblast HTR-8 cells.MethodsCircRNA and miRNA levels were examined by quantitative reverse transcription PCR (qRT-PCR). Overexpression plasmid constructs and siRNAs were used to overexpress and knockdown hsa_circ_0081343, respectively. Transwell assays and flow cytometry analyses were performed to evaluate the effects of hsa_circ_0081343 or miR-210-5p on migration, invasion, and apoptosis. Protein levels were analyzed by western blotting. Dual luciferase activity and anti-AGO2 RNA immunoprecipitation (RIP) assays were performed to identify the relationship between miR-210-5p and hsa_circ_0081343.ResultsHsa_circ_0081343 expression was significantly downregulated in 37 FGR placental tissues compared to healthy placental control tissues. Hsa_circ_0081343 overexpression may inhibit apoptosis by downregulating the expression of cleaved caspase 3 and caspase 9 and alleviating the migration and invasion of HTR-8 cells by inducing the expression of MMP2 and MMP9. The dual luciferase activity and anti-AGO2 RIP assay results showed that hsa_circ_0081343 binds to miR-210-5p. miR-210-5p overexpression eliminated the effect of hsa_circ_0081343 overexpression in HTR-8 cells. Finally, DLX3 was identified as a direct target of miR-210-5p.Conclusionshsa_circ_0081343 expression levels are significantly downregulated in FGR placental tissues. Hsa_circ_0081343 regulates the migration, invasion, and apoptosis of HTR-8 cells via the hsa-miR-210-5p/DLX3 axis.
Highlights
Various circular RNAs are dysregulated in the placenta of fetal growth restriction (FGR) fetuses, but their roles and regulatory mechanisms have not been fully elucidated
Our results showed that hsa_circ_0081343 expression levels were increased in HTR-8 cells upon transfection with ov-circ_0081343 as compared to the cells transfected with the empty vector or negative control (NC) group (Fig. 2A)
SiRNA-2 demonstrated the highest efficacy in silencing the expression of hsa_circ_0081343 at 100 nM when compared to that of the si-NC group (Fig. 2B)
Summary
Various circular RNAs (circRNAs) are dysregulated in the placenta of fetal growth restriction (FGR) fetuses, but their roles and regulatory mechanisms have not been fully elucidated. Fetal growth restriction (FGR) refers to a condition in which the fetus does not maintain its intrauterine potential for growth and development. This condition further results in the fetal birth weight being less than 2500 g after 37 weeks of gestation, which is. FGR is a multifactorial disorder, and the associated placental dysfunction has been linked to the deceleration of fetal growth This dysfunction can result in a reduction in oxygen and nutrient supply from the mother to the fetus [5]. The precise molecular mechanisms underlying placental development and function remain unclear
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