Downregulated lncRNA HOXA11-AS Affects Trophoblast Cell Proliferation and Migration by Regulating RND3 and HOXA7 Expression in PE

Abstract

The long noncoding RNA HOXA11-AS displays abnormal expression in numerous human diseases. However, its function and biological mechanisms remain unclear in preeclampsia (PE). In this study, we report that HOXA11-AS is significantly downregulated in preeclamptic placental tissues and could contribute to the occurrence and development of PE. Silencing of HOXA11-AS expression could significantly suppress trophoblast cell growth and migration, whereas HOXA11-AS overexpression facilitated cell growth in the HTR-8/SVneo, JEG3, and JAR cell lines. RNA-seq analysis also indicated that HOXA11-AS silencing preferentially regulated numerous genes associated with cell proliferation and cell migration. Mechanistic analyses showed that HOXA11-AS could recruit Ezh2 and Lsd1 protein and regulate RND3 mRNA expression in the nucleus. In the cytoplasm, HOXA11-AS modulates HOXA7 expression by sponged miR-15b-5p, affecting trophoblast cell proliferation. Together, these data confirm that aberrant expression of HOXA11-AS is involved in the occurrence and development of PE and may act as a prospective diagnosis and therapeutic target in PE.

Down‐regulated lncRNA AGAP2‐AS1 contributes to pre‐eclampsia as a competing endogenous RNA for JDP2 by impairing trophoblastic phenotype

“…These assays were conducted as previously described by Xu et al After transfecting cells with either siRNAs or the plasmid, we performed fluorescein isothiocyanate (FITC)‐annexin V and propidium iodide (PI) staining using the FITC‐Annexin V Apoptosis Detection kit (BD Biosciences), following the manufacturer's instructions. The cell cycle level was determined by propidium oxide staining using the Cycle Test Plus DNA Reagent kit (BD Biosciences), following the protocol.…”

“…This experiment was carried out as previously described in Xu et al Briefly, HTR‐8/SVneo and JEG3 cells were transfected with specific siRNAs and/or plasmid; they were then re‐suspended in RPMI‐1640 medium containing 1% FBS and subsequently added to the upper chamber. The lower chamber was filled with 700 µl RPMI‐1640 (Gibco) medium containing 10% FBS.…”

“…This experiment was conducted as defined earlier by Xu et al Firstly, HTR‐8/SVneo cells were plated in 6‐well plates. After 24‐48 hours, the cells were fixed with 75% ethanol for 30 minutes and then washed thrice with PBS.…”

“…This experiment was conducted as defined earlier by Xu et al Trophoblast cells were preserved with formaldehyde and incubated for 10 minutes to generate DNA‐protein cross links. Then, cell lysates were sonicated to generate chromatin fragments of 200‐300 bp and immunoprecipitated with FOXP1 (Millipore) or IgG as the control.…”

“…Despite the lack of a general consensus regarding their biological characteristics, there is growing evidence that lncRNAs have a high degree of regulation and important functions in human diseases. Specifically, lncRNAs have emerged as key molecular players in the regulation of gene expression in different biological processes such as parental imprinting, stem cell pluripotency, cell growth, and metastasis and apoptosis . It is considered that these lncRNAs are regulated through effects such as miRNA sponging, chromatin remodelling and epigenetic modification .…”