miR-24-3p Regulates Progression of Gastric Mucosal Lesions and Suppresses Proliferation and Invasiveness of N87 Via Peroxiredoxin 6

Dig Dis Sci. 2016 Dec;61(12):3486-3497. doi: 10.1007/s10620-016-4309-9. Epub 2016 Oct 14.

Abstract

Background: MicroRNAs, targeting mRNAs of cancer-associated genes, are often aberrantly expressed in human gastric cancer (GC).

Aim: We have examined the possible role and mechanisms of miRNA regulation of Prdx-6 in the development and progression of H. pylori-related gastric mucosal lesions.

Methods: First, miR-24-3p was predicted to target Prdx-6, and this negative regulation was validated by luciferase reporter analyses, Western blot, and quantitative RT-PCR. Next, immunohistochemistry and in situ hybridization were performed to detect the Prdx-6 and miR-24-3p expression in tissue microarrays of gastric mucosal lesions. Finally, the miR-24-3p function in GC cell line N87 was examined by MTT, Annexin V-FITC, PI, transwell migration, and Matrigel invasion assays.

Results: In our study, Prdx-6 expression was negatively regulated by miR-24-3p expression and miR-24-3p interacted with the 3'-untranslated region of Prdx-6 to down-regulate its expression level. In addition, miR-24-3p expression gradually decreased in human gastric specimens from chronic superficial gastritis (CSG) to dysplasia and was upregulated in GC tissues compared with adjacent normal tissues. Contrary to this, Prdx-6 expression showed inverse tendency in the same tissue. More so, expression of miR-24-3p was reduced in samples with H. pylori infection, especially in CSG. Moreover, miR-24-3p was associated with GC lymph nodes and liver metastasis. Gain- or loss-of-function experiments showed that miR-24-3p significantly inhibited N87 cell growth, migration, and invasion and promoted apoptosis, while Prdx-6 reversed these miR-24-3p effects.

Conclusions: miR-24-3p was identified as a regulator of development and progression of H. pylori-related gastric mucosal lesions.

Keywords: Gastric cancer; H. pylori; Prdx-6; miR-24.

MeSH terms

  • Apoptosis / genetics
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation / genetics
  • Down-Regulation
  • Gastric Mucosa
  • Gastritis / genetics*
  • Gastritis / metabolism
  • Gastritis / pathology
  • Gene Expression Regulation, Neoplastic*
  • Helicobacter Infections / genetics*
  • Helicobacter Infections / metabolism
  • Helicobacter Infections / pathology
  • Helicobacter pylori
  • Humans
  • Immunohistochemistry
  • In Situ Hybridization
  • MicroRNAs / genetics*
  • Neoplasm Invasiveness
  • Peroxiredoxin VI / genetics*
  • Peroxiredoxin VI / metabolism
  • RNA, Messenger / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stomach Neoplasms / genetics*
  • Stomach Neoplasms / metabolism
  • Stomach Neoplasms / pathology
  • Up-Regulation

Substances

  • MIRN24 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • PRDX6 protein, human
  • Peroxiredoxin VI