Effects of JUN and NFE2L2 knockdown on oxidative status and NFE2L2/AP-1 targets expression in HeLa cells in basal conditions and upon sub-lethal hydrogen peroxide treatment

Mol Biol Rep. 2019 Feb;46(1):27-39. doi: 10.1007/s11033-018-4412-4. Epub 2018 Dec 4.

Abstract

Although NFE2L2 transcription factor is considered to make the most significant contribution to the NFE2L2/AP-1-pathway-dependent antioxidants regulation in the human cell, AP-1 has the potential to provide significant backup and even play an equal role in the cell. Considering this, the present study is focused on revealing how JUN, an AP-1 component, and NFE2L2 contribute to regulation of four target genes containing AREs with embedded TREs-SQSTM1, FTH1, HMOX1 and CBR3 and to cellular oxidative status in general in basal conditions and under pro-oxidative influence. NFE2L2 and JUN were down-regulated in HeLa cells using siRNA-mediated knockdown approach. These cells were subsequently exposed to 400 µM hydrogen peroxide in the medium or equal volume of sterile water. They revealed some evidence of both backup functioning and competing between the two factors. Importantly, JUN demonstrated a high level of participation (inc. as a negative regulator) in functioning of the classic NFE2L2 targets and in cellular oxidative status establishment in general. One of the key findings was a dramatic increase in JUN expression following NFE2L2 knockdown in basal conditions. The both AP-1 and NFE2L2 sub-pathways equally determine the outcome of the NFE2L2/AP-1 pathway activation induced by various stimuli, and the outcome is stimulus type- and stimulus-intensity-specific and results from either of the two eventually dominating sub-pathways.

Keywords: AP-1; Antioxidant signaling; NFE2L2; NRF2; Oxidative status.

MeSH terms

  • Antioxidants / metabolism
  • Down-Regulation / drug effects
  • Epithelial Cells / metabolism
  • Gene Expression Regulation / drug effects
  • Gene Knockdown Techniques / methods
  • HeLa Cells
  • Heme Oxygenase-1 / genetics
  • Humans
  • Hydrogen Peroxide / metabolism
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / physiology*
  • Oxidation-Reduction
  • Oxidative Stress / drug effects
  • Oxidative Stress / genetics*
  • Proto-Oncogene Proteins c-jun / genetics
  • Proto-Oncogene Proteins c-jun / physiology*
  • Transcription Factor AP-1 / genetics
  • Transcription Factor AP-1 / physiology

Substances

  • Antioxidants
  • NF-E2-Related Factor 2
  • NFE2L2 protein, human
  • Proto-Oncogene Proteins c-jun
  • Transcription Factor AP-1
  • Hydrogen Peroxide
  • Heme Oxygenase-1