Casticin impairs cell growth and induces cell apoptosis via cell cycle arrest in human oral cancer SCC-4 cells

Environ Toxicol. 2018 Feb;33(2):127-141. doi: 10.1002/tox.22497. Epub 2017 Nov 3.

Abstract

Casticin, a polymethoxyflavone, present in natural plants, has been shown to have biological activities including anti-cancer activities. Herein, we investigated the anti-oral cancer activity of casticin on SCC-4 cells in vitro. Viable cells, cell cycle distribution, apoptotic cell death, reactive oxygen species (ROS) production, and Ca2+ production, levels of ΔΨm and caspase activity were measured by flow cytometric assay. Cell apoptosis associated protein expressions were examined by Western blotting and confocal laser microscopy. Results indicated that casticin induced cell morphological changes, DNA condensation and damage, decreased the total viable cells, induced G2 /M phase arrest in SCC-4 cells. Casticin promoted ROS and Ca2+ productions, decreases the levels of ΔΨm , promoted caspase-3, -8, and -9 activities in SCC-4 cells. Western blotting assay demonstrated that casticin affect protein level associated with G2/M phase arrest and apoptosis. Confocal laser microscopy also confirmed that casticin increased the translocation of AIF and cytochrome c in SCC-4 cells. In conclusion, casticin decreased cell number through G2 /M phase arrest and the induction of cell apoptosis through caspase- and mitochondria-dependent pathways in SCC-4 cells.

Keywords: SCC-4 human oral cancer cells; apoptosis; caspase- and mitochondria-dependent pathways; casticin.

MeSH terms

  • Apoptosis / drug effects*
  • Calcium / metabolism
  • Caspase 3 / metabolism
  • Caspase 8 / metabolism
  • Caspase 9 / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects*
  • Cytochromes c / metabolism
  • DNA Damage / drug effects
  • Flavonoids / toxicity*
  • G2 Phase Cell Cycle Checkpoints / drug effects*
  • Humans
  • M Phase Cell Cycle Checkpoints / drug effects*
  • Membrane Potential, Mitochondrial / drug effects
  • Microscopy, Confocal
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Mouth Neoplasms / metabolism
  • Mouth Neoplasms / pathology
  • Reactive Oxygen Species / metabolism

Substances

  • Flavonoids
  • Reactive Oxygen Species
  • casticin
  • Cytochromes c
  • Caspase 3
  • Caspase 8
  • Caspase 9
  • Calcium