Hypermethylation of miR-338-3p and Impact of its Suppression on Cell Metastasis Through N-Cadherin Accumulation at the Cell -Cell Junction and Degradation of MMP in Gastric Cancer

Cell Physiol Biochem. 2018;50(2):411-425. doi: 10.1159/000494153. Epub 2018 Oct 11.

Abstract

Background/aims: MicroRNAs (miRNAs) have been well studied in human carcinogenesis and cancer progression. Our previous study showed the down-regulation of miR-338-3p expression in human gastric cancer (GC). However, the reasons of this dysregulation remain largely unclear.

Methods: Bisulfite sequence analysis was performed to explore the methylation status of the promoter region of miR-338-3p. Cell wound-healing and transwell assays were performed to examine the capacity of cell migration and cell interaction. A dual-luciferase reporter was used to validate the bioinformatics-predicted target gene of miR-338-3p. Western blotting, RNA interference, and immunofluorescence (IF) were used to evaluate the expression of MMPs and the location of N-cadherin to determine the mechanism underlying miR-338-3p-induced anti-tumor effects.

Results: miR-338-3p was epigenetically silenced, and this loss of expression was significantly correlated with the Borrmann Stage in GC. Restoring miR-338-3p expression in BGC-823 cells inhibited cell migration and invasion. Moreover, Ras-related protein (Rab-14) and Hedgehog acyltransferase (Hhat) were identified as direct targets of miR-338-3p. Both enforced expression of miR-338-3p and small interfering RNA induced Rab14-mediated accumulation of N-cadherin in the cell -cell junctions or Hhat-associated matrix metalloproteinase (MMP) degradation, which may underline the metastasis defects caused by loss of miR-338-3p in GC.

Conclusion: These data indicate that miR-338-3p functions as a tumor suppressor in GC, and that the hypermethylation status of its CpG island might be a novel potential strategy for treating GC.

Keywords: Gastric cancer; Hhat-MMP signaling; Metastasis; Rab14-N-cadherin; miR-338-3p.

MeSH terms

  • 3' Untranslated Regions
  • Acyltransferases / antagonists & inhibitors
  • Acyltransferases / genetics
  • Acyltransferases / metabolism
  • Antagomirs / metabolism
  • Base Sequence
  • Cadherins / metabolism*
  • Cell Line, Tumor
  • Cell Movement
  • DNA Methylation
  • Humans
  • Intercellular Junctions / metabolism*
  • Matrix Metalloproteinases / metabolism*
  • MicroRNAs / antagonists & inhibitors
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Sequence Alignment
  • Signal Transduction
  • Stomach Neoplasms / genetics
  • Stomach Neoplasms / pathology*
  • rab GTP-Binding Proteins / antagonists & inhibitors
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / metabolism

Substances

  • 3' Untranslated Regions
  • Antagomirs
  • Cadherins
  • MIRN338 microRNA, human
  • MicroRNAs
  • RNA, Small Interfering
  • Acyltransferases
  • HHAT protein, human
  • Matrix Metalloproteinases
  • Rab14 protein, human
  • rab GTP-Binding Proteins