The role of NF-kappaB in 6-hydroxydopamine- and TNFalpha-induced apoptosis of PC12 cells

Naunyn Schmiedebergs Arch Pharmacol. 2004 Jun;369(6):563-9. doi: 10.1007/s00210-004-0938-1. Epub 2004 May 13.

Abstract

6-Hydroxydopamine (6-OHDA) is widely used to study the death of catecholaminergic cells related to Parkinson's disease. Oxidative stress and gene transcription are known to mediate the pro-apoptotic effect of 6-OHDA. As redox mechanisms are involved in activation of the transcription factor NF-kappaB, we studied the role of NF-kappaB in 6-OHDA-induced death of PC12 cells. We stably transfected PC12 cells with a doxycycline-regulated expression vector for the NF-kappaB super-repressor (IkappaBalpha mutated at serine-32 and serine-36, IkappaBalpha-SR). NF-kappaB transcriptional activity was evaluated by transient transfection of an NF-kappaB-driven luciferase reporter gene. Expression of IkappaBalpha-SR inhibited NF-kappaB stimulated by tumor necrosis factor alpha (TNFalpha) and 6-OHDA. Apoptosis was quantified by counting cells with condensed nuclei. IkappaBalpha-SR inhibited apoptosis induced by 6-OHDA but enhanced apoptosis that was triggered by TNFalpha. The converse effects of NF-kappaB could be due to different target genes that are induced in the context of TNFalpha and 6-OHDA stimulation. Indeed, TNFalpha stimulated mRNA accumulation of the anti-apoptotic superoxide dismutase 2 through NF-kappaB whereas 6-OHDA induced mRNA accumulation of the pro-apoptotic c-myc. These data demonstrate that NF-kappaB regulates survival of the neuron-like PC12 cells in a stimulus-specific manner. In the context of 6-OHDA stimulation, NF-kappaB mediates pro-apoptotic effects, suggesting that NF-kappaB signaling could be a target for drug development in Parkinson-related neurodegeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / pharmacology
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Doxycycline / pharmacology
  • I-kappa B Proteins / biosynthesis
  • I-kappa B Proteins / genetics
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / genetics
  • NF-kappa B / physiology*
  • Oxidopamine
  • PC12 Cells
  • Rats
  • Transfection
  • Tumor Necrosis Factor-alpha

Substances

  • I-kappa B Proteins
  • NF-kappa B
  • Nfkbia protein, rat
  • Tumor Necrosis Factor-alpha
  • NF-KappaB Inhibitor alpha
  • Oxidopamine
  • 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
  • Doxycycline