Synthesized Pheophorbide a-mediated photodynamic therapy induced apoptosis and autophagy in human oral squamous carcinoma cells

J Oral Pathol Med. 2013 Jan;42(1):17-25. doi: 10.1111/j.1600-0714.2012.01187.x. Epub 2012 Jun 28.

Abstract

Background: Pheophorbide a (Pa) is a chlorine-based photosensitizer derived from an ethnopharmacological herb, and our group recently synthesized Pa by the removal of a magnesium ion and a phytyl group from chlorophyll-a. In this study, the effect of photodynamic therapy (PDT) with synthesized Pa was examined in a human oral squamous cell carcinoma (OSCC) cells.

Methods: Cells were treated with PDT with Pa, and reactive oxygen species (ROS) and mitochondrial membrane potential [ΔΨ (m)] were examined. Apoptosis was measured using annexin V staining and immunoblot. Autophagy was characterized by the increase in LC3B-II and the formation of autophagosome and acidic vesicular organelles (AVOs).

Results: Pa-PDT inhibited the proliferation of OSCC cells in a dose-dependent manner. Pa-PDT increased the number of apoptotic cells by inactivating ERK pathway. Pa-PDT also induced autophagy in OSCC cells evidenced by the increased levels of LC3 type II expression and the accumulation of AVOs. The inhibition of autophagy enhanced Pa-PDT-mediated cytotoxicity through an increase in necrosis.

Conclusions: These results suggest that synthesized Pa-PDT exerts anti-tumor effects by inducing apoptosis and autophagy and provide novel evidence that Pa-PDT induces autophagy, and autophagy inhibition enhances Pa-PDT-mediated necrosis in OSCC cells.

MeSH terms

  • Apoptosis*
  • Autophagy* / drug effects
  • Carcinoma, Squamous Cell / drug therapy*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Chlorophyll / analogs & derivatives*
  • Chlorophyll / chemical synthesis
  • Chlorophyll / pharmacology
  • Chlorophyll / therapeutic use
  • Humans
  • MAP Kinase Signaling System / drug effects
  • Microtubule-Associated Proteins / biosynthesis
  • Mouth Neoplasms / drug therapy*
  • Necrosis
  • Phagosomes
  • Photochemotherapy*
  • Photosensitizing Agents / chemical synthesis
  • Photosensitizing Agents / pharmacology
  • Photosensitizing Agents / therapeutic use*
  • Reactive Oxygen Species / metabolism

Substances

  • MAP1LC3B protein, human
  • Microtubule-Associated Proteins
  • Photosensitizing Agents
  • Reactive Oxygen Species
  • Chlorophyll
  • pheophorbide a