Dimethyl fumarate induces necroptosis in colon cancer cells through GSH depletion/ROS increase/MAPKs activation pathway

Br J Pharmacol. 2015 Aug;172(15):3929-43. doi: 10.1111/bph.13184. Epub 2015 Jun 12.

Abstract

Background and purpose: Dimethyl fumarate (DMF) is a newly approved drug for the treatment of relapsing forms of multiple sclerosis and relapsing-remitting multiple sclerosis. Here, we investigated the effects of DMF and its metabolites mono-methylfumarate (MMF and methanol) on different gastrointestinal cancer cell lines and the underlying molecular mechanisms involved.

Experimental approach: Cell viability was measured by the MTT or CCK8 assay. Protein expressions were measured by Western blot analysis. LDH release, live- and dead-cell staining, intracellular GSH levels, and mitochondrial membrane potential were examined by using commercial kits.

Key results: DMF but not MMF induced cell necroptosis, as demonstrated by the pharmacological tool necrostatin-1, transmission electron microscopy, LDH and HMGB1 release in CT26 cells. The DMF-induced decrease in cellular GSH levels as well as cell viability and increase in reactive oxygen species (ROS) were inhibited by co-treatment with GSH and N-acetylcysteine (NAC) in CT26 cells. DMF activated JNK, p38 and ERK MAPKs in CT26 cells and JNK, p38 and ERK inhibitors partially reversed the DMF-induced decrease in cell viability. GSH or NAC treatment inhibited DMF-induced JNK, p38, and ERK activation in CT26 cells. DMF but not MMF increased autophagy responses in SGC-7901, HCT116, HT29 and CT26 cancer cells, but autophagy inhibition did not prevent the DMF-induced decrease in cell viability.

Conclusion and implications: DMF but not its metabolite MMF induced necroptosis in colon cancer cells through a mechanism involving the depletion of GSH, an increase in ROS and activation of MAPKs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Colonic Neoplasms / enzymology
  • Colonic Neoplasms / metabolism
  • Colonic Neoplasms / pathology*
  • Dimethyl Fumarate / adverse effects*
  • Fumarates / adverse effects
  • Glutathione / metabolism*
  • HMGB1 Protein / metabolism
  • Humans
  • L-Lactate Dehydrogenase / metabolism
  • MAP Kinase Signaling System / drug effects*
  • Maleates / adverse effects
  • Membrane Potential, Mitochondrial / drug effects
  • Methanol / adverse effects
  • Mice
  • Mitogen-Activated Protein Kinases / metabolism*
  • Necrosis / chemically induced*
  • Necrosis / metabolism
  • Reactive Oxygen Species / metabolism*

Substances

  • Fumarates
  • HMGB1 Protein
  • Maleates
  • Reactive Oxygen Species
  • citraconic acid
  • L-Lactate Dehydrogenase
  • Mitogen-Activated Protein Kinases
  • Dimethyl Fumarate
  • Glutathione
  • Methanol