Influence of hydrogen-occluding-silica on migration and apoptosis in human esophageal cells in vitro

Abstract

In the last decade, many studies have shown that hydrogen gas or hydrogen water can reduce the levels of reactive oxygen species in the living body. Molecular hydrogen has antioxidant and antiapoptotic effects and a preventive effect on oxidative stress-induced cell death. In the present study, we investigated solidified hydrogen-occluding-silica (H2-silica) that can release molecular hydrogen into cell culture medium because the use of hydrogen gas has strict handling limitations in hospital and medical facilities and laboratories, owing to its physicochemical characteristics. Human esophageal squamous cell carcinoma (KYSE-70) cells and normal human esophageal epithelial cells (HEEpiCs) were used to investigate the effects of H2-silica on cell viability and proliferation. Cell migration was examined with wound healing and culture-insert migration assays. The intracellular levels of reactive oxygen species were evaluated with a nitroblue tetrazolium assay. To assess the apoptotic status of the cells, the Bax/Bcl-2 ratio and cleaved caspase-3 were analyzed by western blot. The results showed that KYSE-70 cells and HEEpiCs were generally inhibited by H2-silica administration, and there was a significant proliferation-inhibitory effect in an H2-silica concentration-dependent manner compared with the control group (P < 0.05) in KYSE-70. Apoptosis-inducing effect on KYSE-70 cells was observed in 10, 300, 600, and 1,200 ppm H2-silica, and only 1,200 ppm H2-silica caused a 2.4-fold increase in apoptosis in HEEpiCs compared with the control group as the index of Bax/Bcl-2. H2 silica inhibited cell migration in KYSE-70 cells, and high concentrations had a cytotoxic effect on normal cells. These findings should provide insights into the mechanism of inhibition of H2-silica on human cancer cells in vitro.

Hydrogen: A Novel Option in Human Disease Treatment

“…Interestingly, Wang et al recently discovered that H 2 inhibited the growth, migration, and invasion of the A549 and H1975 lung cancer cell lines and promoted cell apoptosis, suggesting that H 2 might play crucial roles in the treatment of lung cancer [ 44 ]. Li et al also revealed the apoptosis-inducing effect of H 2 on KYSE-70 human esophageal squamous cell carcinoma cells [ 45 ]. Thus, H 2 may protect normal cells from damage and suppressing cancer cells.…”

Effects of hydrogen-occluding-silica microparticles on wound repair and cell migratory behavior of normal human esophageal epitheliocytes

“…In our previous studies, fascin exerted an important function in cell migration and filopodia formation. 10 Since microfilaments exert important actions in processes such as cell migration and pseudopodia formation, formation of filopodia and microvilli can be used as an indicator of cell proliferation. In this study, cell migration was promoted, and formation of microvilli was activated in the 100 ppm (low concentration) scratched group, where a proliferation effect was seen.…”

“…As previously described, 10 HEEpiCs treated with H 2 -silica were seeded at approximately 1 × 10 6 cells per T25 flask and further cultured for 72-96 hours. Cells were collected using a small rubber scraper (TR9000, TrueLine, Nashville, TN, USA), centrifuged at 5000 r/min at 4°C for 5 minutes, then washed twice in ice-cold PBS (–).…”

“…Additional procedures were the same as reported in our previous study. 10 Anti-Bax (BioLegend, CA, USA, #625901), anti-Bcl-2 (eBioscience, CA, #141028), anti-α-tubulin, and anti-fascin-1 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, #SC-21743) were utilized as the primary antibodies, and Jackson Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (#115-035-003) 2.0 mL was utilized as the secondary antibody. Finally, both the ratio of the intensity of activated caspase-3 versus native caspase-3 and the ratio of the intensity of Bax versus Bcl-2 were used to evaluate the occurrence of apoptosis in HEEpiCs.…”

“…In our previous study, the effect of H 2 -silica on cell migration was measured, and the fact was found that H 2 -silica is more effective in inhibiting migration of cancer cells than normal cells. 10 It suggests that H 2 -silica can inhibit the metastasis of cancer cells in vitro , it also suggests that damaged normal cells would be restored and wound-healing effect would appear at the same time.…”