Bax/Bak-independent mitochondrial depolarization and reactive oxygen species induction by sorafenib overcome resistance to apoptosis in renal cell carcinoma

J Biol Chem. 2017 Apr 21;292(16):6478-6492. doi: 10.1074/jbc.M116.754184. Epub 2017 Feb 1.

Abstract

Renal cell carcinoma (RCC) is polyresistant to chemo- and radiotherapy and biologicals, including TNF-related apoptosis-inducing ligand (TRAIL). Sorafenib, a multikinase inhibitor approved for the treatment of RCC, has been shown to sensitize cancer cells to TRAIL-induced apoptosis, in particular by down-regulation of the Bak-inhibitory Bcl-2 family protein Mcl-1. Here we demonstrate that sorafenib overcomes TRAIL resistance in RCC by a mechanism that does not rely on Mcl-1 down-regulation. Instead, sorafenib induces rapid dissipation of the mitochondrial membrane potential (ΔΨm) that is accompanied by the accumulation of reactive oxygen species (ROS). Loss of ΔΨm and ROS production induced by sorafenib are independent of caspase activities and do not depend on the presence of the proapoptotic Bcl-2 family proteins Bax or Bak, indicating that both events are functionally upstream of the mitochondrial apoptosis signaling cascade. More intriguingly, we find that it is sorafenib-induced ROS accumulation that enables TRAIL to activate caspase-8 in RCC. This leads to apoptosis that involves activation of an amplification loop via the mitochondrial apoptosis pathway. Thus, our mechanistic data indicate that sorafenib bypasses central resistance mechanisms through a direct induction of ΔΨm breakdown and ROS production. Activation of this pathway might represent a useful strategy to overcome the cell-inherent resistance to cancer therapeutics, including TRAIL, in multiresistant cancers such as RCC.

Keywords: B cell lymphoma 2 (Bcl-2) family; Bak; Bax; Trail; apoptosis; cancer; drug resistance; reactive oxygen species (ROS); sorafenib.

MeSH terms

  • Apoptosis
  • Carcinoma, Renal Cell / drug therapy
  • Carcinoma, Renal Cell / metabolism*
  • Caspase 8 / metabolism
  • Cell Line, Tumor
  • Down-Regulation
  • Drug Resistance, Neoplasm*
  • Enzyme Activation
  • Flow Cytometry
  • Humans
  • Kidney Neoplasms / drug therapy
  • Kidney Neoplasms / metabolism*
  • Membrane Potential, Mitochondrial
  • Mitochondria / metabolism*
  • Myeloid Cell Leukemia Sequence 1 Protein / metabolism
  • Niacinamide / analogs & derivatives*
  • Niacinamide / pharmacology
  • Phenylurea Compounds / pharmacology*
  • Protein Conformation
  • RNA, Small Interfering / metabolism
  • Reactive Oxygen Species / metabolism*
  • Signal Transduction
  • Sorafenib
  • TNF-Related Apoptosis-Inducing Ligand / metabolism
  • bcl-2 Homologous Antagonist-Killer Protein / metabolism
  • bcl-2-Associated X Protein / metabolism

Substances

  • BAK1 protein, human
  • BAX protein, human
  • MCL1 protein, human
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Phenylurea Compounds
  • RNA, Small Interfering
  • Reactive Oxygen Species
  • TNF-Related Apoptosis-Inducing Ligand
  • TNFSF10 protein, human
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-2-Associated X Protein
  • Niacinamide
  • Sorafenib
  • CASP8 protein, human
  • Caspase 8