Heat shock exerts anticancer effects on liver cancer via autophagic degradation of aquaporin 5

Kudou,Michihiro; Shiozaki,Atsushi; Kosuga,Toshiyuki; Shimizu,Hiroki; Ichikawa,Daisuke; Konishi,Hirotaka; Morimura,Ryo; Komatsu,Shuhei; Ikoma,Hisashi; Fujiwara,Hitoshi; Okamoto,Kazuma; Marunaka,Yoshinori; Otsuji,Eigo

doi:10.3892/ijo.2017.3940

Heat shock exerts anticancer effects on liver cancer via autophagic degradation of aquaporin 5

Authors:
- Michihiro Kudou
- Atsushi Shiozaki
- Toshiyuki Kosuga
- Hiroki Shimizu
- Daisuke Ichikawa
- Hirotaka Konishi
- Ryo Morimura
- Shuhei Komatsu
- Hisashi Ikoma
- Hitoshi Fujiwara
- Kazuma Okamoto
- Yoshinori Marunaka
- Eigo Otsuji
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Affiliations: Division of Digestive Surgery, Department of Surgery, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan, Departments of Molecular Cell Physiology and Bio-Ionomics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan
Published online on: March 29, 2017 https://doi.org/10.3892/ijo.2017.3940
Pages: 1857-1867

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Abstract

Previous studies described that the expression of aquaporin 5 (AQP5) was altered in tumors of various organs. AQP5 is attracting attention as a new cancer therapeutic target. In the present study, heat shock-induced changes in AQP5 expression were evaluated by immunofluorescent staining (IF) and western blotting (WB) of liver cancer cells. AQP5 knockdown experiments or a heat shock treatment were conducted, and their effects on cell volume, proliferation, cell cycle, the activity of apoptosis and migration/invasion were compared. Cycloheximide (CHX) chase experiments and double IF of AQP5 and light chain 3B (LC3B) were performed to investigate the mechanisms underlying changes in AQP5 expression. The results showed that IF and WB revealed decrease in AQP5 expression on cellular membranes and in the cytoplasm of heated cells. AQP5 knockdown and heat shock similarly decreased cell volume, suppressed migration/invasion and proliferation, and induced early apoptosis and partial G0/G1 arrest. CHX chase experiments revealed that heat shock accelerated the degradation of AQP5, which was rescued under CHX and the autophagy inhibitor, bafilomycin A1 (BafA1). Double IF showed the co-localization of AQP5 and LC3B on BafA1-treated heated cells. In conclusion, we demonstrated that heat shock decreased AQP5 on cellular membranes and in the cytoplasm by activating autophagic degradation, and heat shock and AQP5 knockdown exerted similar anticancer effects, suggesting that heat shock exerts anticancer effects via the autophagic degradation of AQP5.

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