Diallyl Disulfide Inhibits Gastric Cancer By Generating Hydrogen Sulfide And Its Preliminary Mechanism

Objective:Diallyl disulfide（DADS）is the main components of fat-soluble substances in garlic.The study found that DADS can inhibit a variety of tumors and has the advantage of low toxicity.It is an anti-tumor drug with great development potential.Our previous study confirmed that DADS can inhibit the proliferation of human gastric cancer cells,block gastric cancer cells in G2/M phase,and inhibit EMT of human gastric cancer cells.Hydrogen sulfide（H₂S）is the third important endogenous gas messenger molecule in the human body after NO and CO.It has been proved that H₂S has a protective effect on gastric mucosa and can inhibit the growth of cancer.DADS is a sulfur-containing compound and may be a donor of H₂S.Therefore,we explored whether DADS can inhibit the growth of gastric cancer cells by producing H₂S and preliminary study its mechanism.Methods:In this study,we used the methylene blue method hydrogen sulfide kits and fluorescence probe experiments were detect to content of human gastric cancer SGC-7901 cells in each treatment group.Immunofluorescence was used to detect the localization of CSE and CBS in cells.CCK8,Transwell migration,Transwell invasion,scratch experiment and flow cytometry were used to detect the effects of cell proliferation,migration,invasion and apoptosis on gastric cancer cell function.After adding CSE inhibitor PPG,observe the changes of each treatment group.Western Blot was used to detect the expression of related proteins in different cells and different treatment groups.Thus,to study the situation of H₂S produced by DADS and the effect of H₂S on gastric cancer cells.Results:1.The methylene blue method and fluorescent probe experiments have proved that DADS can produce H₂S in SGC-7901 cells,and the amount of H₂S produced is related to the concentration of DADS.2.After DADS and NaHS were added to SGC-7901 cells,part of the cell morphology of DADS and NaHS group changed from polygon to circle or oval.3.Immunofluorescence found that CSE and CBS are mainly expressed in the nucleus.4.Western Blot was used to detect the expression of CSE and CBS in gastric mucosa GES-1 and gastric cancer SGC-7901,MGC-803 cells and the expression of gastric cancer SGC-7901 in different treatment groups.Among them,the expression of CSE and CBS in gastric mucosal cell lines was higher than that in gastric cancer cell lines.Among gastric cancer SGC-7901 cells,CSE expression was highest in the DADS group and lowest in the NaHS group.5.Fluorescence probe showed that the content of H₂S in the cells decreased significantly after adding PPG.6.Western Blot and immunofluorescence test results confirm that PPG can significantly reduce the increase of CSE caused by DADS.7.H₂S can inhibit the proliferation,migration,invasion and promote the apoptosis of SGC-7901 cells.8.DADS can inhibit the proliferation,migration,invasion and promote the apoptosis of gastric cancer cells.PPG,an inhibitor of H₂S production,can reduce the above effects of DADS.9.Western Blot detected the expression of EMT-associated pathway proteins in each treatment group.The results showed that the expression levels of E-Cadherin and Claudin-1 in the DADS group and NaHS group increased,and the expression of Slug decreased.Conclusion:1.DADS can produce H₂S in gastric cancer SGC-7901 cells,and the amount of H₂S produced is related to the concentration of DADS.2.H₂S can inhibit the proliferation,migration,invasion and promote the apoptosis of SGC-7901 cells.3.DADS can play an anti-gastric cancer effect by generating H₂S,and its mechanism is related to the inhibition of EMT.