Inhibition of ARC decreases the survival of HEI-OC-1 cells after neomycin damage in vitro

Oncotarget. 2016 Oct 11;7(41):66647-66659. doi: 10.18632/oncotarget.11336.

Abstract

Hearing loss is a common sensory disorder mainly caused by the loss of hair cells (HCs). Noise, aging, and ototoxic drugs can all induce apoptosis in HCs. Apoptosis repressor with caspase recruitment domain(ARC) is a key factor in apoptosis that inhibits both intrinsic and extrinsic apoptosis pathways; however, there have been no reports on the role of ARC in HC loss in the inner ear. In this study, we used House Ear Institute Organ of Corti 1 (HEI-OC-1) cells, which is a cochlear hair-cell-like cell line, to investigate the role of ARC in aminoglycoside-induced HC loss. ARC was expressed in the cochlear HCs as well as in the HEI-OC-1 cells, but not in the supporting cells, and the expression level of ARC in HCs was decreased after neomycin injury in both cochlear HCs and HEI-OC-1 cells, suggesting that reduced levels of ARC might correlate with neomycin-induced HC loss. We inhibited ARC expression using siRNA and found that this significantly increased the sensitivity of HEI-OC-1 cells to neomycin toxicity. Finally, we found that ARC inhibition increased the expression of pro-apoptotic factors, decreased the mitochondrial membrane potential, and increased the level of reactive oxygen species (ROS) after neomycin injury, suggesting that ARC inhibits cell death and apoptosis in HEI-OC-1 cells by controlling mitochondrial function and ROS accumulation. Thus the endogenous anti-apoptotic factor ARC might be a new therapeutic target for the prevention of aminoglycoside-induced HC loss.

Keywords: apoptosis; cochlea; hair cell; mitochondrial function; reactive oxygen species.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Apoptosis Regulatory Proteins / genetics*
  • Apoptosis Regulatory Proteins / metabolism
  • Cell Line
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Cells, Cultured
  • Hair Cells, Auditory / cytology
  • Hair Cells, Auditory / drug effects*
  • Hair Cells, Auditory / metabolism
  • Membrane Potential, Mitochondrial / drug effects
  • Mice
  • Muscle Proteins / genetics*
  • Muscle Proteins / metabolism
  • Neomycin / pharmacology*
  • Protein Synthesis Inhibitors / pharmacology
  • RNA Interference*
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / genetics

Substances

  • Apoptosis Regulatory Proteins
  • Muscle Proteins
  • Nol3 protein, mouse
  • Protein Synthesis Inhibitors
  • Reactive Oxygen Species
  • Neomycin