Beta hydroxybutyrate induces lung cancer cell death, mitochondrial impairment and oxidative stress in a long term glucose-restricted condition

Mol Biol Rep. 2024 Apr 24;51(1):567. doi: 10.1007/s11033-024-09501-w.


Background: Metabolic plasticity gives cancer cells the ability to shift between signaling pathways to facilitate their growth and survival. This study investigates the role of glucose deprivation in the presence and absence of beta-hydroxybutyrate (BHB) in growth, death, oxidative stress and the stemness features of lung cancer cells.

Methods and results: A549 cells were exposed to various glucose conditions, both with and without beta-hydroxybutyrate (BHB), to evaluate their effects on apoptosis, mitochondrial membrane potential, reactive oxygen species (ROS) levels using flow cytometry, and the expression of CD133, CD44, SOX-9, and β-Catenin through Quantitative PCR. The activity of superoxide dismutase, glutathione peroxidase, and malondialdehyde was assessed using colorimetric assays. Treatment with therapeutic doses of BHB triggered apoptosis in A549 cells, particularly in cells adapted to glucose deprivation. The elevated ROS levels, combined with reduced levels of SOD and GPx, indicate that oxidative stress contributes to the cell arrest induced by BHB. Notably, BHB treatment under glucose-restricted conditions notably decreased CD133 expression, suggesting a potential inhibition of cell survival through the downregulation of CD133 levels. Additionally, the simultaneous decrease in mitochondrial membrane potential and increase in ROS levels indicate the potential for creating oxidative stress conditions to impede tumor cell growth in such environmental settings.

Conclusion: The induced cell death, oxidative stress and mitochondria impairment beside attenuated levels of cancer stem cell markers following BHB administration emphasize on the distinctive role of metabolic plasticity of cancer cells and propose possible therapeutic approaches to control cancer cell growth through metabolic fuels.

Keywords: A459 cells; Apoptosis; Beta-hydroxybutyrate; Mitochondria membrane potential (ΔΨm); Reactive oxygen species (ROS); Stemness.

MeSH terms

  • 3-Hydroxybutyric Acid* / pharmacology
  • A549 Cells
  • AC133 Antigen / genetics
  • AC133 Antigen / metabolism
  • Apoptosis* / drug effects
  • Cell Death / drug effects
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Glucose* / metabolism
  • Humans
  • Lung Neoplasms* / drug therapy
  • Lung Neoplasms* / metabolism
  • Lung Neoplasms* / pathology
  • Membrane Potential, Mitochondrial* / drug effects
  • Mitochondria* / drug effects
  • Mitochondria* / metabolism
  • Oxidative Stress* / drug effects
  • Reactive Oxygen Species* / metabolism
  • Superoxide Dismutase / metabolism


  • Glucose
  • 3-Hydroxybutyric Acid
  • Reactive Oxygen Species
  • Superoxide Dismutase
  • AC133 Antigen