Contribution of reactive oxygen species to migration/invasion of human glioblastoma cells U87 via ERK-dependent COX-2/PGE(2) activation

Neurobiol Dis. 2010 Jan;37(1):118-29. doi: 10.1016/j.nbd.2009.09.015. Epub 2009 Oct 3.


In the presence of 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulation, an increase in the migration/invasion of U87 glioblastoma cells was detected by a wound healing assay, transwell analysis, and spheroid formation assay by inducing matrix metalloproteinase-9 (MMP-9) enzyme activity via a gelatin zymographic analysis. A dose- and time-dependent increase in cyclooxygenase-2 (COX-2) gene expression with elevated prostaglandin E(2) (PGE(2)) production was identified in TPA- but not in 4alpha-TPA (a respective inactive compound)-treated U87 cells TPA-induced migration/invasion was significantly blocked by adding the COX-2-specific inhibitor, NS398, through a reduction in PGE(2) production. Data from the pharmacological studies using specific chemical inhibitors showed that activation of protein kinase C (PKC) and extracellular signal-regulated kinases (ERKs) was involved in TPA-induced migration/invasion, COX-2 protein expression, and MMP-9 activation. Stimulation of intracellular peroxide production by TPA was detected by a DCHF-DA assay, and the addition of superoxide dismutase (SOD) or tempol significantly inhibited TPA-induced migration/invasion and COX-2 protein expression accompanied by a decrease in peroxide production. An increase in NADPH oxidase activity by TPA was examined, and TPA-induced migration/invasion was blocked by adding DPI, an NADPH oxidase inhibitor. Additionally, the natural flavonoids quercetin (QE), baicalein (BE), and myricetin (ME) effectively blocked TPA-induced migration/invasion while simultaneously inhibiting COX-2/PGE(2) production, MMP-9 enzyme activity, and peroxide production in U87 cells. The contribution of ROS production to the migration/invasion of U87 glioblastoma cells via ERK-activated COX-2/PGE(2) and MMP-9 induction was first investigated here, and agents such as QE, BE, and ME with the ability to block these events possess the potential to be developed for use against migration/invasion by glioblastomas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Movement / physiology
  • Cyclooxygenase 2 / metabolism*
  • Cyclooxygenase Inhibitors / pharmacology
  • Dinoprostone / metabolism*
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • Glioblastoma / drug therapy
  • Glioblastoma / physiopathology*
  • Humans
  • Intracellular Space / drug effects
  • Intracellular Space / physiology
  • Matrix Metalloproteinase 9 / metabolism*
  • NADPH Oxidases / antagonists & inhibitors
  • NADPH Oxidases / metabolism
  • Peroxides / metabolism
  • Protein Kinase C / metabolism
  • Reactive Oxygen Species / metabolism*
  • Signal Transduction
  • Time Factors


  • Cyclooxygenase Inhibitors
  • Peroxides
  • Reactive Oxygen Species
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • NADPH Oxidases
  • Protein Kinase C
  • Extracellular Signal-Regulated MAP Kinases
  • Matrix Metalloproteinase 9
  • Dinoprostone