Digestive Diseases and Sciences

, Volume 57, Issue 8, pp 2022–2030

Intestinal Epithelial Cells with Impaired Autophagy Lose Their Adhesive Capacity in the Presence of TNF-α

  • Masaya Saito
  • Tatsuro Katsuno
  • Tomoo Nakagawa
  • Toru Sato
  • Yoshiko Noguchi
  • Sayuri Sazuka
  • Keiko Saito
  • Makoto Arai
  • Koutaro Yokote
  • Osamu Yokosuka
Original Article

Abstract

Background and Objectives

Genome-wide association studies have revealed a link between autophagy-related (ATG) genes and susceptibility to Crohn’s disease. This suggests underlying involvement of autophagy impairment in the pathogenesis of Crohn’s disease. This study was performed to investigate the pathophysiological importance of autophagy impairment in intestinal epithelial cells exposed to TNF-α.

Methods

Human colonic epithelial cells (HT-29) and rat small intestinal epithelial cells (IEC-18) were used. Formation of phosphatidylethanolamine-conjugated microtubule-associated protein light chain 3 (LC3-II) was monitored as a marker of autophagy. Autophagy was inhibited using 3-methyladenine or short interfering RNA targeting ATG5 and ATG16L1.

Results

TNF-α treatment elicited a significant dose-dependent increase in LC3-II protein levels, thus autophagy is induced in the presence of TNF-α. Combined autophagy inhibition and TNF-α treatment induced a marked increase in the number of detached cells and a decrease in activated integrin β1 protein levels. Trypan blue staining indicated 70–80 % of the detached cells were alive, suggesting that these cells became detached not because they were killed but because of dysfunction of cellular adhesion.

Conclusions

This is the first study indicating that intestinal epithelial cells with impaired autophagy lose their adhesive capacity in the presence of TNF-α. These observations indicate that impairment of autophagy leads to disruption of the intestinal epithelial cell layers in TNF-α-rich environments.

Keywords

Crohn’s disease Autophagy-related (ATG) gene Phosphatidylethanolamine-conjugated microtubule-associated protein light chain 3 (LC3-II) 3-Methyladenine (3-MA) Integrin β1 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Masaya Saito
    • 1
  • Tatsuro Katsuno
    • 1
  • Tomoo Nakagawa
    • 1
  • Toru Sato
    • 1
  • Yoshiko Noguchi
    • 1
  • Sayuri Sazuka
    • 1
  • Keiko Saito
    • 1
  • Makoto Arai
    • 1
  • Koutaro Yokote
    • 2
  • Osamu Yokosuka
    • 1
  1. 1.Department of Medicine and Clinical Oncology (K1), Graduate School of MedicineChiba UniversityChiba-shiJapan
  2. 2.Department of Clinical Cell Biology and Medicine (F5), Graduate School of MedicineChiba UniversityChibaJapan

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