Chloroquine Sensitizes Nasopharyngeal Carcinoma Cells but Not Nasoepithelial Cells to Irradiation by Blocking Autophagy

PLoS One. 2016 Nov 30;11(11):e0166766. doi: 10.1371/journal.pone.0166766. eCollection 2016.

Abstract

Background: Treatment of nasopharyngeal carcinoma requires the application of high dosages of radiation, leading to severe long-term complications in the majority of patients. Sensitizing tumor cells to radiation could be a means to increase the therapeutic window of radiation. Nasopharyngeal carcinoma cells display alterations in autophagy and blockade of autophagy has been shown to sensitize them against chemotherapy.

Methods: We investigated the effect of chloroquine, a known inhibitor of autophagy, on sensitization against radiation-induced apoptosis in a panel of five nasopharyngeal carcinoma cell lines and a SV40-transformed nasoepithelial cell line. Autophagy was measured by immunoblot of autophagy-related proteins, immunofluorescence of autophagosomic microvesicles and electron microscopy. Autophagy was blocked by siRNA against autophagy-related proteins 3, 5, 6 and 7 (ATG3, ATG5, ATG6 and ATG7).

Results: Chloroquine sensitized four out of five nasopharyngeal cancer cell lines towards radiation-induced apoptosis. The sensitizing effect was based on the blockade of autophagy as inhibition of ATG3, ATG5, ATG6 and ATG7 by specific siRNA could substitute for the effect of chloroquine. No sensitization was seen in nasoepithelial cells.

Conclusion: Chloroquine sensitizes nasopharyngeal carcinoma cells but not nasoepithelial cells towards radiation-induced apoptosis by blocking autophagy. Further studies in a mouse-xenograft model are warranted to substantiate this effect in vivo.

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis / radiation effects*
  • Autophagy / drug effects*
  • Autophagy / genetics*
  • Autophagy-Related Protein 5 / genetics
  • Autophagy-Related Protein 7 / genetics
  • Autophagy-Related Proteins / genetics
  • Beclin-1 / genetics
  • Carcinoma / drug therapy
  • Carcinoma / radiotherapy*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Proliferation / radiation effects
  • Chloroquine / pharmacology*
  • Epithelial Cells / drug effects
  • Epithelial Cells / radiation effects*
  • Humans
  • In Situ Nick-End Labeling
  • Nasal Mucosa / cytology
  • Nasopharyngeal Carcinoma
  • Nasopharyngeal Neoplasms / drug therapy
  • Nasopharyngeal Neoplasms / radiotherapy*
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Radiation-Sensitizing Agents / pharmacology*
  • Ubiquitin-Conjugating Enzymes / genetics

Substances

  • ATG5 protein, human
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Beclin-1
  • RNA, Small Interfering
  • Radiation-Sensitizing Agents
  • Chloroquine
  • Ubiquitin-Conjugating Enzymes
  • ATG7 protein, human
  • Autophagy-Related Protein 7
  • ATG3 protein, human

Grants and funding

The authors received no specific funding for this work.