Mechanism of neem limonoids-induced cell death in cancer: Role of oxidative phosphorylation

Free Radic Biol Med. 2016 Jan:90:261-71. doi: 10.1016/j.freeradbiomed.2015.11.028. Epub 2015 Nov 25.

Abstract

We have previously reported that neem limonoids (neem) induce multiple cancer cell death pathways. Here we dissect the underlying mechanisms of neem-induced apoptotic cell death in cancer. We observed that neem-induced caspase activation does not require Bax/Bak channel-mediated mitochondrial outer membrane permeabilization, permeability transition pore, and mitochondrial fragmentation. Neem enhanced mitochondrial DNA and mitochondrial biomass. While oxidative phosphorylation (OXPHOS) Complex-I activity was decreased, the activities of other OXPHOS complexes including Complex-II and -IV were unaltered. Increased reactive oxygen species (ROS) levels were associated with an increase in mitochondrial biomass and apoptosis upon neem exposure. Complex-I deficiency due to the loss of Ndufa1-encoded MWFE protein inhibited neem-induced caspase activation and apoptosis, but cell death induction was enhanced. Complex II-deficiency due to the loss of succinate dehydrogenase complex subunit C (SDHC) robustly decreased caspase activation, apoptosis, and cell death. Additionally, the ablation of Complexes-I, -III, -IV, and -V together did not inhibit caspase activation. Together, we demonstrate that neem limonoids target OXPHOS system to induce cancer cell death, which does not require upregulation or activation of proapoptotic Bcl-2 family proteins.

Keywords: Apoptosis; Mitochondrial DNA; Necroptosis; Neem; Oxidative phosphorylation complex; ROS.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Azadirachta / chemistry*
  • Caspases / metabolism
  • Cyclin-Dependent Kinase Inhibitor p21 / physiology
  • DNA, Mitochondrial / analysis
  • Dynamins
  • Electron Transport Complex I / physiology
  • GTP Phosphohydrolases / analysis
  • HCT116 Cells
  • Humans
  • Limonins / pharmacology*
  • Microtubule-Associated Proteins / analysis
  • Mitochondrial Membrane Transport Proteins / physiology
  • Mitochondrial Permeability Transition Pore
  • Mitochondrial Proteins / analysis
  • Neoplasms / drug therapy
  • Neoplasms / pathology*
  • Oxidative Phosphorylation*
  • Tumor Suppressor Protein p53 / physiology

Substances

  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • DNA, Mitochondrial
  • Limonins
  • Microtubule-Associated Proteins
  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Permeability Transition Pore
  • Mitochondrial Proteins
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Caspases
  • GTP Phosphohydrolases
  • DNM1L protein, human
  • Dynamins
  • Electron Transport Complex I