Nox1-dependent superoxide production controls colon adenocarcinoma cell migration

Biochim Biophys Acta. 2008 Jan;1783(1):23-33. doi: 10.1016/j.bbamcr.2007.10.010. Epub 2007 Oct 30.


Reactive oxygen species are well-known mediators of various biological responses. Recently, new homologues of the catalytic subunit of NADPH oxidase have been discovered in non-phagocytic cells. These new homologues (Nox1-Nox5) produce low levels of superoxides compared to the phagocytic homologue Nox2/gp91phox. Using Nox1 siRNA, we show that Nox1-dependent superoxide production affects the migration of HT29-D4 colonic adenocarcinoma cells on collagen-I. Nox1 inhibition or down-regulation led to a decrease of superoxide production and alpha 2 beta 1 integrin membrane availability. An addition of arachidonic acid stimulated Nox1-dependent superoxide production and HT29-D4 cell migration. Pharmacological evidences using phospholipase A2, lipoxygenases and protein kinase C inhibitors show that upstream regulation of Nox1 relies on arachidonic acid metabolism. Inhibition of 12-lipoxygenase decreased basal and arachidonic acid induced Nox1-dependent superoxide production and cell migration. Migration and ROS production inhibited by a 12-lipoxygenase inhibitor were restored by the addition of 12(S)-HETE, a downstream product of 12-lipoxygenase. Protein kinase C delta inhibition by rottlerin (and also GO6983) prevented Nox1-dependent superoxide production and inhibited cell migration, while other protein kinase C inhibitors were ineffective. We conclude that Nox1 activation by arachidonic acid metabolism occurs through 12-lipoxygenase and protein kinase C delta, and controls cell migration by affecting integrin alpha 2 subunit turn-over.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / metabolism*
  • Adenocarcinoma / pathology*
  • Arachidonate 12-Lipoxygenase / metabolism
  • Arachidonic Acids / pharmacology
  • Cell Membrane / metabolism
  • Cell Movement* / drug effects
  • Colonic Neoplasms / metabolism*
  • Colonic Neoplasms / pathology*
  • HT29 Cells
  • Humans
  • Integrin alpha2beta1 / metabolism
  • NADPH Oxidase 1
  • NADPH Oxidases / metabolism*
  • Protein Kinase C-delta / metabolism
  • Superoxides / antagonists & inhibitors
  • Superoxides / metabolism*


  • Arachidonic Acids
  • Integrin alpha2beta1
  • Superoxides
  • Arachidonate 12-Lipoxygenase
  • NADPH Oxidase 1
  • NADPH Oxidases
  • NOX1 protein, human
  • Protein Kinase C-delta