Naringin Combined with NF-κB Inhibition and Endoplasmic Reticulum Stress Induces Apoptotic Cell Death via Oxidative Stress and the PERK/eIF2α/ATF4/CHOP Axis in HT29 Colon Cancer Cells

Biochem Genet. 2021 Feb;59(1):159-184. doi: 10.1007/s10528-020-09996-5. Epub 2020 Sep 26.


Currently, combination therapy is considered the most effective solution for a selective chemotherapeutic effect in the treatment of colon cancer. This study investigated the death of both colon cancer HT29 cells and healthy vascular smooth muscle TG-Ha-VSMC cells (VSMCs) induced by naringin combined with endoplasmic reticulum (ER) stress and NF-κB inhibition. Naringin combined with tunicamycin and BAY 11-7082 suppressed the proliferation of HT29 cells in a dose-dependent manner and induced particularly apoptotic death without significantly affecting healthy VSMCs according to Annexin V/PI staining and AO/EB staining analyses. Insufficient antioxidant defense and heat shock response as well as excessive ROS generation were observed in HT29 cells following combination therapy. Quantitative real-time PCR and western blot analysis demonstrated that drug combination-induced mitochondrial apoptosis was activated through the ROS-mediated PERK/eIF2α/ATF4/CHOP pathway. Additionally, naringin combination significantly reduced the sXBP expression induced by tunicamycin+BAY 11-7082 in a dose-dependent manner. In conclusion, this study found that naringin combined with tunicamycin+BAY 11-7082 efficiently induced apoptotic cell death in HT29 colon cancer cells via oxidative stress and the PERK/eIF2α/ATF4/CHOP pathway, suggesting that naringin combined with tunicamycin plus BAY 11-7082 could be a new combination therapy strategy for effective colon cancer treatment with minimal side effects on healthy cells.

Keywords: Apoptosis; BAY 11-7082; Colon cancer; ER stress; Naringin; Tunicamycin.

MeSH terms

  • Activating Transcription Factor 4 / metabolism
  • Antioxidants / pharmacology
  • Apoptosis*
  • Caspases / metabolism
  • Cell Line, Tumor
  • Cell Survival
  • Colonic Neoplasms / drug therapy
  • Endoplasmic Reticulum / metabolism
  • Endoplasmic Reticulum Stress* / drug effects
  • Eukaryotic Initiation Factor-2 / metabolism
  • Flavanones / administration & dosage
  • Flavanones / pharmacology*
  • HT29 Cells
  • Humans
  • Mitochondria / metabolism
  • Muscle, Smooth, Vascular / cytology
  • NF-kappa B p50 Subunit / antagonists & inhibitors*
  • NF-kappa B p50 Subunit / metabolism
  • Nitriles / pharmacology
  • Oxidative Stress*
  • Reactive Oxygen Species
  • Signal Transduction*
  • Sulfones / pharmacology
  • Transcription Factor CHOP / metabolism
  • Tunicamycin / administration & dosage
  • eIF-2 Kinase / metabolism


  • 3-(4-methylphenylsulfonyl)-2-propenenitrile
  • ATF4 protein, human
  • Antioxidants
  • DDIT3 protein, human
  • Eukaryotic Initiation Factor-2
  • Flavanones
  • NF-kappa B p50 Subunit
  • NFKB1 protein, human
  • Nitriles
  • Reactive Oxygen Species
  • Sulfones
  • Tunicamycin
  • Activating Transcription Factor 4
  • Transcription Factor CHOP
  • EIF2AK3 protein, human
  • eIF-2 Kinase
  • Caspases
  • naringin