Suppressive effects of electrochemically reduced water on matrix metalloproteinase-2 activities and in vitro invasion of human fibrosarcoma HT1080 cells

• Published in: Cytotechnology (Dordrecht) 2012-05, Vol.64 (3), p.357-371
• Main Authors: Kinjo, Tomoya, Ye, Jun, Yan, Hanxu, Hamasaki, Takeki, Nakanishi, Hidekazu, Toh, Kazuko, Nakamichi, Noboru, Kabayama, Shigeru, Teruya, Kiichiro, Shirahata, Sanetaka
• Format: Article
• Language: English
• Subjects: Acids; Biochemistry; Biological and medical sciences; Biomedicine; Biotechnology; Cancer; Cells; Chemistry; Chemistry and Materials Science; Diabetes; Down-regulation; Electrolysis; Enzyme inhibitors; Extracellular matrix; Fibrosarcoma; Fundamental and applied biological sciences. Psychology; Gelatinase A; Gene expression; Hydrogen; Hydrogen peroxide; Kinases; MAP kinase; Matrix metalloproteinase; Metalloproteinase; Metastasis; Nanoparticles; Nematodes; Original Research; Oxidative stress; Phosphorylation; Signal transduction; Transcription factors; Tumors
• ISSN: 0920-9069; 1573-0778
• DOI: 10.1007/s10616-012-9469-7

It has been demonstrated that hydrogen peroxide (H 2 O 2 ) is directly associated with elevated matrix metalloproteinase-2 (MMP-2) expression in several cell lines. Electrochemically reduced water (ERW), produced near the cathode during electrolysis, and scavenges intracellular H 2 O 2 in human fibrosarcoma HT1080 cells. RT-PCR and zymography analyses revealed that when HT1080 cells were treated with ERW, the gene expression of MMP-2 and membrane type 1 MMP and activation of MMP-2 was repressed, resulting in decreased invasion of the cells into matrigel. ERW also inhibited H 2 O 2 -induced MMP-2 upregulation. To investigate signal transduction involved in MMP-2 downregulation, mitogen-activated protein kinase (MAPK)-specific inhibitors, SB203580 (p38 MAPK inhibitor), PD98059 (MAPK/extracellular regulated kinase kinase 1 inhibitor) and c-Jun NH 2 -terminal kinase inhibitor II, were used to block the MAPK signal cascade. MMP-2 gene expression was only inhibited by SB203580 treatment, suggesting a pivotal role of p38 MAPK in regulation of MMP-2 gene expression. Western blot analysis showed that ERW downregulated the phosphorylation of p38 both in H 2 O 2 -treated and untreated HT1080 cells. These results indicate that the inhibitory effect of ERW on tumor invasion is due to, at least in part, its antioxidative effect.