Repeated H2 O2 exposure drives cell cycle progression in an in vitro model of ulcerative colitis

J Cell Mol Med. 2013 Dec;17(12):1619-31. doi: 10.1111/jcmm.12150. Epub 2013 Oct 9.

Abstract

The production of hydrogen peroxide (H2 O2 ) drives tumourigenesis in ulcerative colitis (UC). Recently, we showed that H2 O2 activates DNA damage checkpoints in human colonic epithelial cells (HCEC) through c-Jun N-terminal Kinases (JNK) that induces p21(WAF1) . Moreover, caspases circumvented the G1/S and intra-S checkpoints, and cells accumulated in G2/M. The latter observation raised the question of whether repeated H2 O2 exposures alter JNK activation, thereby promoting a direct passage of cells from G2/M arrest to driven cell cycle progression. Here, we report that increased proliferation of repeatedly H2 O2 -exposed HCEC cells (C-cell cultures) was associated with (i) increased phospho-p46 JNK, (ii) decreased total JNK and phospho-p54 JNK and (iii) p21(WAF1) down-regulation. Altered JNK activation and p21(WAF1) down-regulation were accompanied by defects in maintaining G2/M and mitotic spindle checkpoints through adaptation, as well as by apoptosis resistance following H2 O2 exposure. This may cause increased proliferation of C-cell cultures, a defining initiating feature in the inflammation-carcinoma pathway in UC. We further suggest that dysregulated JNK activation is attributed to a non-apoptotic function of caspases, causing checkpoint adaptation in C-cell cultures. Additionally, loss of cell-contact inhibition and the overcoming of senescence, hallmarks of cancer, contributed to increased proliferation. Furthermore, there was evidence that p54 JNK inactivation is responsible for loss of cell-contact inhibition. We present a cellular model of UC and suggest a sinusoidal pattern of proliferation, which is triggered by H2 O2 -induced reactive oxygen species generation, involving an interplay between JNK activation/inactivation, p21(WAF1) , c-Fos, c-Jun/phospho-c-Jun, ATF2/phospho-ATF2, β-catenin/TCF4-signalling, c-Myc, CDK6 and Cyclin D2, leading to driven cell cycle progression.

Keywords: DNA damage checkpoint adaptation; cell cycle progression; hydrogen peroxide; ulcerative colitis.

MeSH terms

  • Caspases / metabolism
  • Cell Cycle / drug effects*
  • Cell Cycle Checkpoints / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Colitis, Ulcerative / pathology*
  • Contact Inhibition / drug effects
  • Cyclin D2 / metabolism
  • Cyclin-Dependent Kinase 6 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Down-Regulation / drug effects
  • Enzyme Activation / drug effects
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / enzymology
  • Humans
  • Hydrogen Peroxide / pharmacology*
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • Models, Biological*
  • Reactive Oxygen Species / metabolism
  • Transcription Factors / metabolism

Substances

  • Cyclin D2
  • Cyclin-Dependent Kinase Inhibitor p21
  • Reactive Oxygen Species
  • Transcription Factors
  • Hydrogen Peroxide
  • Cyclin-Dependent Kinase 6
  • JNK Mitogen-Activated Protein Kinases
  • Caspases