Induction of Endoplasmic Reticulum Stress via Reactive Oxygen Species Mediated by Luteolin in Melanoma Cells

Anticancer Res. 2016 May;36(5):2281-9.

Abstract

Background: This study aimed to investigate whether luteolin, a flavonoid, induces apoptosis in human melanoma cells via endoplasmic reticulum (ER) stress.

Materials and methods: To investigate the effects of luteolin in human melanoma cells, the anti-proliferation, apoptosis, ER stress induction and reactive oxygen species (ROS) generation were evaluated using MTT, Hoechst 33342, ER-tracker Blue White DPX and DCF-DA staining assays, respectively.

Results: Luteolin inhibited cell proliferation and increased apoptotic body formation. Luteolin induced ER stress, as shown by ER staining and mitochondrial Ca(2+) overloading. Luteolin increased expression of the ER stress-related proteins; protein kinase RNA-like ER kinase, phospho eukaryotic translation initiation factor 2α, activating transcription factor (ATF) 6, CCAAT/enhancer-binding protein-homologous protein (CHOP), and cleaved caspase 12. Furthermore, luteolin increased the level of intracellular ROS, leading to ROS-mediated apoptosis and ER stress. However, N-acetyl cysteine, a ROS scavenger, decreased ROS levels, apoptosis, and ER stress induced by luteolin treatment. In addition, knockdown of CHOP and ATF6 by small-interfering RNA inhibited luteolin-induced cell death.

Conclusion: Luteolin induces apoptosis by ER stress via increasing ROS levels.

Keywords: Ca2+ overloading; ER stress; Luteolin; Reactive oxygen species; melanoma.

MeSH terms

  • Activating Transcription Factor 6 / antagonists & inhibitors
  • Activating Transcription Factor 6 / genetics
  • Apoptosis / drug effects
  • Calcium / analysis
  • Cell Line, Tumor
  • Endoplasmic Reticulum Stress / drug effects*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Luteolin / pharmacology*
  • Melanoma / pathology*
  • Mitochondria / chemistry
  • Mitochondria / drug effects
  • Neoplasm Proteins / antagonists & inhibitors
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • RNA, Small Interfering / genetics
  • Reactive Oxygen Species*
  • Staining and Labeling
  • Transcription Factor CHOP / antagonists & inhibitors
  • Transcription Factor CHOP / genetics
  • Transfection
  • Tumor Stem Cell Assay

Substances

  • ATF6 protein, human
  • Activating Transcription Factor 6
  • DDIT3 protein, human
  • Neoplasm Proteins
  • RNA, Small Interfering
  • Reactive Oxygen Species
  • Transcription Factor CHOP
  • Luteolin
  • Calcium