Anoxia/reoxygenation induces epithelial-mesenchymal transition in human colon cancer cell lines

Okajima,Manabu; Kokura,Satoshi; Ishikawa,Takeshi; Mizushima,Katsura; Tsuchiya,Reiko; Matsuyama,Tatsuzo; Adachi,Satoko; Okayama,Tetsuya; Sakamoto,Naoyuki; Kamada,Kazuhiro; Katada,Kazuhiro; Uchiyama,Kazuhiko; Handa,Osamu; Takagi,Tomohisa; Yagi,Nobuaki; Naito,Yuji; Yoshikawa,Toshikazu

doi:10.3892/or.2013.2401

Anoxia/reoxygenation induces epithelial-mesenchymal transition in human colon cancer cell lines

Authors:
- Manabu Okajima
- Satoshi Kokura
- Takeshi Ishikawa
- Katsura Mizushima
- Reiko Tsuchiya
- Tatsuzo Matsuyama
- Satoko Adachi
- Tetsuya Okayama
- Naoyuki Sakamoto
- Kazuhiro Kamada
- Kazuhiro Katada
- Kazuhiko Uchiyama
- Osamu Handa
- Tomohisa Takagi
- Nobuaki Yagi
- Yuji Naito
- Toshikazu Yoshikawa
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Affiliations: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan, Department of Cancer Immuno Cell Regulation, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan, Department of Cancer Immuno Cell Regulation, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan
Published online on: April 10, 2013 https://doi.org/10.3892/or.2013.2401
Pages: 2311-2317

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Abstract

Epithelial-mesenchymal transition (EMT) is considered to be a crucial event in the development of cancer metastasis. Anoxia/reoxygenation (A/R) is known to occur in cancer tissues due to angiogenesis and changes in tissue pressure that occur during tumor growth. We investigated whether A/R induces EMT in the human colon cancer cell line HT-29. Colon cancer cells were exposed to anoxia (2 h) followed by reoxygenation (4-22 h) and evaluated for EMT changes using immunofluorescence and western blot analyses. We also investigated the expression of EMT-related transcription factors (Snail and ZEB1) using RT-PCR and evaluated the expression of NF-κB using ELISA. To determine whether NF-κB is involved in A/R-induced EMT, HT-29 cells were treated with proteasome inhibitors. Colon cancer cells exposed to A/R underwent EMT morphological changes; the cancer cells acquired a spindle-shaped phenotype. The expression of E-cadherin on the cell surface and the total amount of E-cadherin proteins were reduced after A/R. The expression of EMT-related transcription factors (Snail, ZEB1) was increased after A/R. Pretreatment with proteasome inhibitors significantly attenuated the downregulation of E-cadherin induced by A/R. These results indicate that A/R induces EMT in human colon cancer cells through an NF-κB-dependent transcriptional pathway.

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