THE INHIBITION OF MMP-2 EXPRESSION IN HUMAN FIBROSARCOMA HT1080 CELLS BY ELECTROLYZED^EDUCED WATER

摘要

It is known that hydrogen peroxide (H_2O_2) is involved in tumor metastasis and matrix metalloproteinase2 (MMP- 2) expression can be activated by extracellular H2O2. Electrolyzed-reduced water (ERW) produced near the cathode during electrolysis of water exhibits high pH, low dissolved oxygen, extremely high dissolved molecular hydrogen, and extremely negative redox potential values. We expected that ERW which acts as an antagonizing factor to scavenge H_2O_2, would attenuate MMP-2 gsne expression. The RT-PCR analysis revealed that when human fibrosarcoma HT1Q80 cells were treated with ERW for 24h, 48h, 72h, the expression MMP-2 was decreased. The invasion of the cells on matrigel was also repressed. To further investigate the signal transduction involved in the down-regulation of MMP-2, MAPK-specific inhibitors were used to block the MAPK signal cascade underlying the process of MMP-2 expression. HT1080 cells were treated with 10 μM SB203580 (inhibitor for p38 MAPK), 20 μM PD98059 (inhibitor forMEK-1), 40 nM JNKi (inhibitor for JNK) for 24 h, respectively. The MMP-2 expression was inhibited only by the SB203580 treatment. In order to further elucidate the mechanism underlying the down-regulation of MMP induced by the ERW treatment, the promoterof MMP-2 was isolated, and the luciferase assay was performed. The results indicated that the down-regulation of MMP-2 after the ERW treatment was due to the inhibition of the bindings of transcriptional factors on the promoter of MMP-2, which might have been activated by p38 MAPK signal cascade.