Over-expression of nm23-H1 in HeLa cells provides cells with higher resistance to oxidative stress possibly due to raising intracellular p53 and GPX1

Acta Pharmacol Sin. 2008 Dec;29(12):1451-8. doi: 10.1111/j.1745-7254.2008.00902.x.

Abstract

Aim: To determine whether the antitumor factor nm23 is related with antioxidation.

Methods: Full-length human nm23-H1 was cloned into a mammalianexpressing vector and transiently introduced into HeLa cells.

Results: A remarkably low level of reactive oxygen species (ROS) was detected in the cells overexpressing nm23-H1. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays found that the cells transfected with a nm23- H1-expressing plasmid had higher viability and stronger resistance to oxidative stress. Immunoprecipitation tests revealed that endogenous nm23-H1 formed a protein complex with p53. Furthermore, the intracellular levels of p53 and p53- regulated gene GPX1 were obviously increased in the cells overexpressing nm23- H1. The downregulation of p53 in the cells overexpressing nm23-H1 resulted in a higher cellular ROS level and lower cell viability.

Conclusion: The findings suggest that nm23-H1 may act as a cellular protector against oxidative stress, possibly triggering the p53-related antioxidative pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Glutathione Peroxidase / genetics
  • Glutathione Peroxidase / metabolism*
  • Glutathione Peroxidase GPX1
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / metabolism
  • NM23 Nucleoside Diphosphate Kinases / genetics
  • NM23 Nucleoside Diphosphate Kinases / metabolism*
  • Oxidants / metabolism
  • Oxidative Stress*
  • Reactive Oxygen Species / metabolism
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • NM23 Nucleoside Diphosphate Kinases
  • Oxidants
  • Reactive Oxygen Species
  • Tumor Suppressor Protein p53
  • Hydrogen Peroxide
  • Glutathione Peroxidase
  • NME1 protein, human
  • Glutathione Peroxidase GPX1
  • GPX1 protein, human