Over-expression of long non-coding RNA NORAD promotes trophoblastic cell viability, migration, and invasion in preeclampsia via the miR-202-5p/FXR1 axis

Taiwan J Obstet Gynecol. 2022 Mar;61(2):255-264. doi: 10.1016/j.tjog.2022.02.012.

Abstract

Objective: Abnormal expression of long non-coding RNAs (lncRNAs) is critical in preeclampsia (PE) pathogenesis. The current study explored the function of non-coding RNA activated by DNA damage (NORAD) in the progression of PE.

Materials and methods: Quantitative real-time PCR was utilized to determine the expression of NORAD, microRNA (miR)-202-5p, and fragile X-related gene 1 (FXR1) in PE and normal placenta tissues. Cell viability was determined using 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-h-tetrazolium bromide assay, and cell migration and invasion were assessed using the Transwell assay. Target relationships were confirmed with the dual-luciferase reporter assay. Western blotting was performed to determine the protein level of FXR1.

Results: NORAD expression was markedly reduced in PE placenta. NORAD over-expression enhanced the viability, migration, and invasion of HTR-8/SVneo cells. miR-202-5p was a target and was negatively regulated by NORAD. Down-regulation of miR-202-5p promoted the viability, migration, and invasion of HTR-8/SVneo cells. miR-202-5p inversely regulated FXR1 expression by targeting the 3'UTR of FXR1. Both miR-202-5p up-regulation and FXR1 knockdown reversed the NORAD over-expression-induced enhancement in the viability, migration, and invasion of HTR-8/SVneo cells.

Conclusion: Collectively, the results revealed that NORAD over-expression promoted trophoblast viability, invasion, and migration by regulating the miR-202-5p/FXR1 axis. These findings clarify PE pathogenesis and will inform the discovery of new targets for PE treatment.

Keywords: Fragile X-related gene 1; MiR-202-5p; NORAD; Preeclampsia; Trophoblast.

MeSH terms

  • Cell Survival / genetics
  • Female
  • Humans
  • MicroRNAs* / genetics
  • Pre-Eclampsia* / genetics
  • Pre-Eclampsia* / metabolism
  • Pregnancy
  • RNA, Long Noncoding* / genetics
  • RNA, Long Noncoding* / metabolism
  • RNA-Binding Proteins* / genetics
  • RNA-Binding Proteins* / metabolism
  • Trophoblasts / metabolism

Substances

  • FXR1 protein, human
  • MIRN202 microRNA, human
  • MicroRNAs
  • NORAD long non-coding RNA, human
  • RNA, Long Noncoding
  • RNA-Binding Proteins