Butyrate-induced cell death and differentiation are associated with distinct patterns of ROS in HT29-derived human colon cancer cells

Dig Dis Sci. 2010 Apr;55(4):920-30. doi: 10.1007/s10620-009-0820-6. Epub 2009 May 12.


To investigate the role of reactive oxygen species (ROS) induced by butyrate in tumor cells, we compared HT29R, an HT29-derived human colon cancer cell line refractory to butyrate-induced cell differentiation but highly sensitive to cell death, with the differentiation-positive HT29-12 and HT29-21 cell lines (exhibiting low sensitivity to butyrate-induced cell death), with respect to levels of butyrate-induced free radicals (FRs), ROS, and H(2)O(2). Dose-dependent increase of FRs (as determined by electron spin resonance spectroscopy) and ROS (dichlorofluorescein assay) was induced in HT29R, but not in HT29-12 and HT29-21 cells, where, in contrast to HT29R, a dose-dependent increase of H(2)O(2) release (phenol red assay) was induced by butyrate. The mode of butyrate-induced cell death in HT29R cells was of a mixed type with necrosis predominating, which, however, switched to apoptosis as the major type of cell death in the presence of the drugs 1,5-dihydroxyisoquinoline, resveratrol, or cyclosporine A. The results suggest that FRs and ROS induced by butyrate in HT29R cells are products of cell death, while H(2)O(2) induced in HT29-12 and HT29-21 cells is functionally related to cell differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / pharmacology
  • Apoptosis / drug effects
  • Butyrates / pharmacology*
  • Cell Adhesion / drug effects
  • Cell Death / drug effects*
  • Cell Differentiation / drug effects*
  • Cell Division / drug effects
  • Cyclosporine / pharmacology
  • Dose-Response Relationship, Drug
  • Electron Spin Resonance Spectroscopy
  • Free Radicals
  • HT29 Cells
  • Humans
  • Hydrogen Peroxide / metabolism
  • Immunosuppressive Agents / pharmacology
  • In Situ Nick-End Labeling
  • Isoquinolines / pharmacology
  • Necrosis
  • Reactive Oxygen Species / metabolism*
  • Resveratrol
  • Stilbenes / pharmacology


  • Antioxidants
  • Butyrates
  • Free Radicals
  • Immunosuppressive Agents
  • Isoquinolines
  • Reactive Oxygen Species
  • Stilbenes
  • 1,5-dihydroxyisoquinoline
  • Cyclosporine
  • Hydrogen Peroxide
  • Resveratrol