title: Inhibitory effect of electrolyzed reduced water on tumor angiogenesis.
abstract: 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 H(2)O(2) and decreased the release of H(2)O(2) 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.
authors.affiliation: Graduate School of Systems Life Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ye
authors.forename: Yuping
authors.initials: Y
authors.lastname: Li
authors.forename: Takeki
authors.initials: T
authors.lastname: Hamasaki
authors.forename: Noboru
authors.initials: N
authors.lastname: Nakamichi
authors.forename: Takaaki
authors.initials: T
authors.lastname: Komatsu
authors.forename: Taichi
authors.initials: T
authors.lastname: Kashiwagi
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Teruya
authors.forename: Ryuhei
authors.initials: R
authors.lastname: Nishikawa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kawahara
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Osada
authors.forename: Kazuko
authors.initials: K
authors.lastname: Toh
authors.forename: Masumi
authors.initials: M
authors.lastname: Abe
authors.forename: Huaize
authors.initials: H
authors.lastname: Tian
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kabayama
authors.forename: Kazumichi
authors.initials: K
authors.lastname: Otsubo
authors.forename: Shinkatsu
authors.initials: S
authors.lastname: Morisawa
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Katakura
authors.forename: Sanetaka
authors.initials: S
authors.lastname: Shirahata
mesh_terms: D002478:Cells, Cultured; D004572:Electrolysis; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D009369:Neoplasms / Q000098:blood supply*; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010084:Oxidation-Reduction; D012333:RNA, Messenger / Q000032:analysis; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D014867:Water
chemical_list: D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D014867:Water; D006861:Hydrogen Peroxide; D048049:Extracellular Signal-Regulated MAP Kinases