Circular RNA hsa_circ_0000848 Promotes Trophoblast Cell Migration and Invasion and Inhibits Cell Apoptosis by Sponging hsa-miR-6768-5p

Abstract

Background: Fetal growth restriction (FGR) is a worldwide problem, and a major cause of perinatal morbidity. The precise molecular mechanisms involved in placental development and function during FGR remain poorly understood. Circular RNAs (circRNAs) are important biological molecules associated with disease pathogenesis. However, the role of circRNAs in FGR has not been well studied. Methods: circRNA expression profiles in placental tissues with and without FGR were identified by circRNA microarray. circRNA expression was verified by quantitative reverse-transcription PCR (RT-qPCR) assay. The effect of hsa_circ_0000848 and hsa-miR-6768-5p on HTR-8 cell apoptosis, migration, and invasion was evaluated. The association between hsa_circ_0000848 and hsa-miR-6768-5p was confirmed by dual luciferase activity and anti-AGO2 RNA immunoprecipitation (RIP) assays. Protein levels were examined via western blotting. Results: RT-qPCR results showed that hsa_circ_0000848 expression was significantly down-regulated in FGR placenta. Hsa_circ_0000848 overexpression and hsa-miR-6768-5p inhibitor suppressed apoptosis, and promoted cell migration and invasion. In addition, hsa_circ_0000848 overexpression and hsa-miR-6768-5p inhibitor increased the protein abundance of BCL2, MMP2 and MMP9, and decreased the protein abundance of cleaved caspase-3, cleaved caspase-9, and BAX, whereas hsa_circ_0000848 knockdown caused the opposite effect. Moreover, a significant increase in hsa-miR-6768-5p expression and a negative correlation between hsa_circ_0000848 and hsa-miR-6768-5p were identified in the FGR tissues. Luciferase reporter and RIP assay results revealed binding of hsa-miR-6768-5p to hsa_circ_0000848. Furthermore, hsa-miR-6768-5p overexpression eliminated the effect of hsa_circ_0000848 overexpression in HTR-8 cells.

Circular RNA CircCOL5A1 Sponges the MiR-7-5p/Epac1 Axis to Promote the Progression of Keloids Through Regulating PI3K/Akt Signaling Pathway

“…Therefore, the changes of total protein and phosphorylated protein levels of PI3K and Akt were detected by silencing or overexpressing circRNA and miRNA. In fact, many studies on the circRNA pathway have adopted this approach, that is, the expression of the terminal pathway was verified by regulating the circRNA/miRNA/mRNA axis ( Luo et al, 2020 ; Wang et al, 2020a ). Taken together, this evidence supported that PI3K/Akt signaling pathway functioned in accelerating full-thickness wound healing and attenuating keloid formation.…”

CircAMOTL1 Promotes Tumorigenesis Through miR-526b/SIK2 Axis in Cervical Cancer

“…Non-coding RNAs have been manifested not to be the transcriptional noise, and but to be the important regulatory molecules ( Zhuang et al, 2015 ; Chen et al, 2016 , 2017 ; Li et al, 2016a , b ; Tran et al, 2020 ), and circRNAs are novel endogenous non-coding RNAs that have been identified as owing key regulatory roles in the cancer biology ( Ma et al, 2019 ; Di Agostino et al, 2020 ; Dong et al, 2020 ; Jin et al, 2020 ; Li and Wang, 2020 ; Liu et al, 2020 ; Ou et al, 2020 ; Zhang et al, 2020a ). circRNAs act as important roles in the development and progression of cancers and are involved in various biological processes, such as cell proliferation, apoptosis, and migration by regulating gene expression ( Liu et al, 2019 ; Di Agostino et al, 2020 ; Ji et al, 2020 ; Tang and Chen, 2020 ; Wang et al, 2020c ). Mangy circRNAs have been reported to play important functions in cervical cancers, such as hsa_circ_0000515 ( Tang and Chen, 2020 ), has_ circ_0000388 ( Meng et al, 2020 ), circ-HIPK3 ( Qian and Huang, 2020 ), circ-ITCH ( Li et al, 2020 ; Wu et al, 2020b ), circCLK3 ( Jin et al, 2020 ), etc.…”

Circular RNA hsa_circ_0081343 promotes trophoblast cell migration and invasion and inhibits trophoblast apoptosis by regulating miR-210-5p/DLX3 axis

“…Thirty-seven pregnant women with FGR and thirty seven healthy pregnant women enrolled in this study were the same as the participants in our previous study [7]. Their detailed characteristics have been reported in our previous study.…”

“…After 24 h of transfection, the Transwell assays were performed to examine the migration and invasion capabilities of the indicated groups using the same method as described in our previous study [7]. The migrated or invaded cells at the bottom of the lter membrane were photographed using a light microscope (Nikon, Tokyo, Japan) at 200X magni cation.…”

“…In our previous study, we found that the expression pro les of numerous circular RNAs (circRNAs), a class of single-stranded molecules [6], were dysregulated in the placenta of FGR fetuses [7]. Moreover, we found that hsa_circ_0000848 is involved in regulating migration, invasion, and apoptosis of trophoblast cells, suggesting that it may play a role in the etiology and pathogenesis of FGR [7]. CircRNAs exist widely in mammalian cells and are involved in the regulation of gene expression at the transcriptional or posttranscriptional levels [8,9].…”