Oestrogen-related receptor alpha inverse agonist XCT-790 arrests A549 lung cancer cell population growth by inducing mitochondrial reactive oxygen species production

Cell Prolif. 2010 Apr;43(2):103-13. doi: 10.1111/j.1365-2184.2009.00659.x.

Abstract

Objective: Although oestrogen-related receptor alpha (ERRalpha) is primarily thought to regulate energy homeostasis, it also serves as a prognostic marker for cancer. The aim of this study was to investigate any connection between ERRalpha activity and cell population growth.

Materials and methods: XCT-790, an ERRa specific inverse agonist, was employed to suppress ERRa activity in human non-small cell lung cancer cells (NSCLC) A549. Gene expressions were detected using quantitative real-time PCR and Western blot analysis. Mitochondrial mass, membrane potential and reactive oxygen species (ROS) production were measured by staining with Mitotracker green, JC-1 and CM-H(2)DCFDA dyes respectively. Rate of progression through the tricarboxylic acid (TCA) cycle was analysed by measuring activities of citrate synthase and succinate dehydrogenase. Cell cycle analysis was performed by using flow cytometry.

Results: We found that XCT-790 treatment reduced mitochondrial mass but enhanced mitochondrial ROS production by increasing rate through the TCA cycle, elevating mitochondrial membrane potential (DeltaPsi(m)) and down-regulating expression of superoxide dismutase. It was further demonstrated that XCT-790-induced ROS modulated p53 and Rb signalling pathways and suppressed cell replication.

Conclusions: ERRalpha affects cell cycle mechanisms through modulating mitochondrial mass and function. Dysregulation of this essential pathway leads to elevation in mitochondrial ROS production, which in turn modulates activities of tumour suppressors, resulting in cell cycle arrest.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzimidazoles / metabolism
  • Carbocyanines / metabolism
  • Carcinoma, Non-Small-Cell Lung / metabolism*
  • Carcinoma, Non-Small-Cell Lung / pathology
  • Cell Cycle
  • Cell Line, Tumor
  • Dose-Response Relationship, Drug
  • ERRalpha Estrogen-Related Receptor
  • Fluorescent Dyes / metabolism
  • Humans
  • Lung / metabolism
  • Lung / pathology
  • Lung Neoplasms / metabolism*
  • Lung Neoplasms / pathology
  • Membrane Potential, Mitochondrial / drug effects
  • Membrane Potentials / drug effects
  • Mitochondria / drug effects
  • Nitriles / pharmacology*
  • Reactive Oxygen Species / analysis
  • Reactive Oxygen Species / metabolism*
  • Receptors, Estrogen / agonists*
  • Signal Transduction / drug effects
  • Superoxide Dismutase / metabolism
  • Thiazoles / pharmacology*
  • Time Factors
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Benzimidazoles
  • Carbocyanines
  • Fluorescent Dyes
  • Nitriles
  • Reactive Oxygen Species
  • Receptors, Estrogen
  • Thiazoles
  • Tumor Suppressor Protein p53
  • XCT790
  • 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine
  • Superoxide Dismutase