MicroRNA-34a induces apoptosis in the human glioma cell line, A172, through enhanced ROS production and NOX2 expression

Biochem Biophys Res Commun. 2014 Jan 31;444(1):6-12. doi: 10.1016/j.bbrc.2013.12.136. Epub 2014 Jan 4.

Abstract

Background: MicroRNA is a type of non-coding small RNA involved in regulating genes and signaling pathways through incomplete complementation with target genes. Recent research supports key roles of miRNA in the formation and development of human glioma.

Methods: The relative quantity of miR-34a was initially determined in human glioma A172 cells and glioma tissues. Next, we analyzed the impact of miR-34a on A172 cell viability with the MTT assay. The effects of miR-34a overexpression on apoptosis were confirmed with flow cytometry and Hoechst staining experiments. We further defined the target genes of miR-34a using immunofluorescence and Western blot.

Results: MiR-34a expression was significantly reduced in human glioma A172 cells and glioma tissue, compared with normal glial cells and tissue samples. Our MTT data suggest that up-regulation of miR-34a inhibits cell viability while suppression of miR-34a enhances cell viability. Flow cytometry and Hoechst staining results revealed increased rates of apoptosis in A172 human glioma cells overexpressing miR-34a. Using immunofluorescence and Western blot analyses, we identified NOX2 as a target of miR-34a in A172 cells.

Conclusion: MiR-34a serves as a tumor suppressor in human glioma mainly by decreasing NOX2 expression.

Keywords: Apoptosis; Human glioma cell line A172; NOX2; miR-34a.

MeSH terms

  • Apoptosis / genetics*
  • Apoptosis / physiology
  • Cell Line, Tumor
  • Cell Survival
  • Central Nervous System Neoplasms / genetics
  • Central Nervous System Neoplasms / metabolism
  • Central Nervous System Neoplasms / pathology
  • Down-Regulation
  • Glioma / genetics*
  • Glioma / metabolism*
  • Glioma / pathology
  • Humans
  • Membrane Glycoproteins / antagonists & inhibitors
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism*
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism*
  • NADPH Oxidase 2
  • NADPH Oxidases / antagonists & inhibitors
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism*
  • Neuroglia / metabolism
  • RNA, Neoplasm / genetics
  • RNA, Neoplasm / metabolism
  • RNA, Small Interfering / genetics
  • Reactive Oxygen Species / metabolism

Substances

  • MIRN34 microRNA, human
  • Membrane Glycoproteins
  • MicroRNAs
  • RNA, Neoplasm
  • RNA, Small Interfering
  • Reactive Oxygen Species
  • CYBB protein, human
  • NADPH Oxidase 2
  • NADPH Oxidases