Clathrin-mediated endocytosis is required for ANE 30-100K-induced autophagy

J Oral Pathol Med. 2018 Jan;47(1):25-31. doi: 10.1111/jop.12593. Epub 2017 Jun 21.

Abstract

Background: We identified an autophagy-inducing areca nut (AN) ingredient (AIAI) in the 30-100 kDa fraction of AN extract (ANE 30-100K). This study was to analyze the role of endocytosis in ANE 30-100K-induced autophagy.

Methods: We used benzyl alcohol, dynasore, and shRNA of clathrin and dynamin to assess whether ANE 30-100K-induced cytotoxicity and accumulation of microtubule-associated protein 1 light chain 3 (LC3)-II were affected in oral (OECM-1) and esophageal (CE81T/VGH) carcinoma cells.

Results: Both benzyl alcohol and dynasore effectively reduced ANE 30-100K-induced cytotoxicity and LC3-II accumulation in OECM-1 and CE81T/VGH cells. Downregulated protein expression of both clathrin and dynamin by their shRNA also significantly attenuated ANE 30-100K-induced elevation of LC3-II levels in CE81T/VGH cells.

Conclusions: These results indicate that AIAI may be engulfed by cells through clathrin-mediated endocytosis, which promotes the execution of the following autophagy program.

Keywords: areca nut; autophagy; benzyl alcohol; clathrin; dynamin; dynasore; endocytosis.

MeSH terms

  • Areca / chemistry*
  • Autophagy / drug effects*
  • Benzyl Alcohol / pharmacology
  • Cell Line, Tumor / drug effects
  • Clathrin / pharmacology*
  • Down-Regulation / drug effects
  • Endocytosis / drug effects*
  • Humans
  • Hydrazones / pharmacology
  • Microtubule-Associated Proteins / metabolism
  • Mouth Neoplasms / chemically induced*
  • Nuts / chemistry
  • Plant Extracts / chemistry
  • Plant Extracts / pharmacology*
  • RNA, Small Interfering / metabolism

Substances

  • Clathrin
  • Hydrazones
  • MAP1LC3B protein, human
  • Microtubule-Associated Proteins
  • N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide
  • Plant Extracts
  • RNA, Small Interfering
  • Benzyl Alcohol