ALKBH5 promotes invasion and metastasis of gastric cancer by decreasing methylation of the lncRNA NEAT1

J Physiol Biochem. 2019 Aug;75(3):379-389. doi: 10.1007/s13105-019-00690-8. Epub 2019 Jul 9.

Abstract

N6-Methyladenosine (m6A) is the most common posttranscriptional modification of RNA and plays critical roles in cancer pathogenesis. However, the biological function of long noncoding RNA (lncRNA) methylation remains unclear. As a demethylase, ALKBH5 (alkylation repair homolog protein 5) is involved in mediating methylation reversal. The purpose of this study was to investigate lncRNA m6A modification and its role in gastric cancer (GC). Bioinformatics predicted interactions of ALKBH5 with lncRNAs. Five methods were employed to assess the function of nuclear paraspeckle assembly transcript 1 (NEAT1), including gene silencing, RT-PCR, separation of nuclear and cytoplasmic fractions, scrape motility assays, and transwell migration assays. Then, m6A RNA immunoprecipitation and immunofluorescence were used to detect methylated NEAT1 in GC cells. Rescue assays were performed to define the relationship between NEAT1 and ALKBH5. NEAT1 is a potential binding lncRNA of ALKBH5. NEAT1 was overexpressed in GC cells and tissue. Additional experiments confirmed that knockdown of NEAT1 significantly repressed invasion and metastasis of GC cells. ALKBH5 affected the m6A level of NEAT1. The binding of ALKBH5 and NEAT1 influences the expression of EZH2 (a subunit of the polycomb repressive complex) and thus affects GC invasion and metastasis. Our findings indicate a novel mechanism by which ALKBH5 promotes GC invasion and metastasis by demethylating the lncRNA NEAT1. They may be potential therapeutic targets for GC.

Keywords: Alkylation repair homolog protein 5; Gastric cancer; Long noncoding RNAs; N 6-Methyladenosine; Nuclear paraspeckle assembly transcript 1.

MeSH terms

  • AlkB Homolog 5, RNA Demethylase / physiology*
  • Cell Line, Tumor
  • Cell Movement
  • Enhancer of Zeste Homolog 2 Protein / metabolism
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Methylation
  • Neoplasm Invasiveness
  • RNA, Long Noncoding / physiology*
  • Stomach Neoplasms* / metabolism
  • Stomach Neoplasms* / pathology

Substances

  • NEAT1 long non-coding RNA, human
  • RNA, Long Noncoding
  • ALKBH5 protein, human
  • AlkB Homolog 5, RNA Demethylase
  • EZH2 protein, human
  • Enhancer of Zeste Homolog 2 Protein