Inhibitory Effect of Electrolyzed Reduced Water on Tumor Angiogenesis

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  • Ye Jun
    Graduate School of Systems Life Sciences, Kyushu University Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Science, Xiamen University
  • Li Yuping
    Graduate School of Systems Life Sciences, Kyushu University School of Life Sciences, Nanchang University of Science and Technology
  • Hamasaki Takeki
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Nakamichi Noboru
    Functional Water Cell Analysis Center Co., Ltd.
  • Komatsu Takaaki
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Kashiwagi Taichi
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Teruya Kiichiro
    Graduate School of Systems Life Sciences, Kyushu University Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Nishikawa Ryuhei
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Kawahara Takeshi
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Osada Kazuhiro
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Toh Kazuko
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Abe Masumi
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Tian Huaize
    Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Kabayama Shigeru
    Nihon Trim Co., Ltd.
  • Otsubo Kazumichi
    Nihon Trim Co., Ltd.
  • Morisawa Shinkatsu
    Nihon Trim Co., Ltd.
  • Katakura Yoshinori
    Graduate School of Systems Life Sciences, Kyushu University Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
  • Shirahata Sanetaka
    Graduate School of Systems Life Sciences, Kyushu University Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University

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Vascular endothelial growth factor (VEGF) is a key mediator of tumor angiogenesis. Tumor cells are exposed to higher oxidative stress compared to normal cells. Numerous reports have demonstrated that the intracellular redox (oxidation/reduction) state is closely associated with the pattern of VEGF expression. Electrolyzed reduced water (ERW) produced near the cathode during the electrolysis of water scavenged intracellular H2O2 and decreased the release of H2O2 from a human lung adenocarcinoma cell line, A549, and down-regulated both VEGF transcription and protein secretion in a time-dependent manner. To investigate the signal transduction pathway involved in regulating VEGF expression, mitogen-activated kinase (MAPK) specific inhibitors, SB203580 (p38 MAPK inhibitor), PD98059 (ERK1/2 inhibitor) and JNKi (c-Jun N-terminal protein kinase inhibitor) were applied. The results showed that only PD98059 blocks VEGF expression, suggesting an important role for ERK1/2 in regulating VEGF expression in A549 cells. As well, ERW inhibited the activation of extracellular signal-regulated kinase (ERK) in a time-dependent manner. Co-culture experiments to analyze in vitro tubule formation assay revealed that A549 cell-derived conditioned medium significantly stimulated the formation of vascular tubules in all analyzed parameters; tubule total area, tubule junction, number of tubules, and total tubule length. ERW counteracted the effect of A549 cell-conditioned medium and decreased total tube length (p<0.01). The present study demonstrated that ERW down-regulated VEGF gene transcription and protein secretion through inactivation of ERK.

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