Early single cell bifurcation of pro- and antiapoptotic states during oxidative stress

J Biol Chem. 2004 Jun 25;279(26):27494-501. doi: 10.1074/jbc.M312135200. Epub 2004 Apr 12.

Abstract

In a population of cells undergoing oxidative stress, an individual cell either succumbs to apoptotic cell death or maintains homeostasis and survives. Exposure of PC-12-D(2)R cells to 200 microm hydrogen peroxide (H(2)O(2)) induces apoptosis in about half of cells after 24 h. After 1-h exposure to 200 microm H(2)O(2), both antiapoptotic extracellular regulated kinase (ERK) phosphorylation and pro-apoptotic Ser-15-p53 phosphorylation are observed. Microarray and real-time PCR assays of gene expression after H(2)O(2) exposure identified several transcripts, including egr1, that are rapidly induced downstream of ERK. Single cell analysis of egr1 induction and of phospho-ERK and phospho-p53 formation revealed the presence of two distinct cellular programs. Whereas the proportion of cells activating ERK versus p53 at 1 h depended on H(2)O(2) concentration, individual cells showed exclusively either phospho-p53 formation or activation of ERK and egr1 induction. Exposure to H(2)O(2) for 1 h also elicited these two non-overlapping cellular responses in both dopaminergic SN4741 cells and differentiated postmitotic PC-12-D(2)R cells. Repressing p53 with pifithrin-alpha or small interfering RNA increased ERK phosphorylation by H(2)O(2), indicating that p53-dependent suppression of ERK activity may contribute to the bi-stable single cell responses observed. By 24 h, the subset of cells in which ERK activity was suppressed exhibit caspase 3 activation and the nuclear condensation characteristic of apoptosis. These studies suggest that the individual cell rapidly and stochastically processes the oxidative stress stimulus, leading to an all-or-none cytoprotective or pro-apoptotic signaling response.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • Caspase 3
  • Caspases / metabolism
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cell Line
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Enzyme Activation / drug effects
  • Gene Expression Profiling
  • Hydrogen Peroxide / toxicity
  • MAP Kinase Signaling System / physiology
  • Mice
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism
  • Oxidative Stress / drug effects
  • Oxidative Stress / physiology*
  • PC12 Cells
  • Phosphorylation
  • RNA / biosynthesis
  • Rats
  • Reactive Oxygen Species / metabolism*
  • Transcription Factors / biosynthesis
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Reactive Oxygen Species
  • Transcription Factors
  • Tumor Suppressor Protein p53
  • RNA
  • Hydrogen Peroxide
  • Mitogen-Activated Protein Kinases
  • Casp3 protein, mouse
  • Casp3 protein, rat
  • Caspase 3
  • Caspases