Hydrogen Sulfide Upregulation Sirt1Expression And Inhibits Migration Of HepG-2Cells

【Objective】To research of hydrogen sulfide (H2S) on the migration of hepatocellularcarcinoma HepG-2cells migration and Sirt1-dependent mechanism, providing a newtarget for the prevention and treatment of hepatocellular carcinoma.【Methods】1. Cultured hepatocellular carcinoma HepG-2cells in vitro, set up control groupand100,200and400μmol/L of H2S donor sodium hydrosulfide (NaHS), a total of4groups. Treated with HepG-2cells after16h, effect of H2S on HepG-2cells healingability were detection by Wound Healing assay; after24h, effect of H2S on HepG-2cells migration ability were detection by Transwell assay; and by Western blotanalysis to the effects of H2S on MMP-9and Sirt1expression of in HepG-2cells.2. Take the most obvious group which NaHS to inhibit HepG-2cell migration,set up blank control group and NaHS (400μmol/L), Sirtinol (Sirt1inhibitors,20μmol/L)+NaHS (400μmol/L) and Sirtinol (20μmol/L), a total of4groups. Pre-treament of HepG-2cells with20μmol/L Sirtinol after30min, inhibiting Sirt1expression of HepG-2cells, and then add400μmol/L NaHS were common treatedwith HepG-2cells after16h; effect of H2S on HepG-2cells healing ability weredetection by Wound Healing assay; after24h, effect of H2S on HepG-2cellsmigration ability were detection by Transwell assay; the expression of MMP-9in HepG-2cells was detection by Western blot.【Results】1.1Wound Healing assay showed that the scuffing distance of HepG-2cells exposedto100,200and400μmol/L H2S donor NaHS were156.8±14.32,139.4±20.21,124.8±14.15μm respectively, significantly decreased compared with174.2±18.99μmof no-treatment group (P＜0.05). Transwell assay showed that the number of HepG2cells migrated through the Matrigel treated by100,200and400μmol/L H2S donorNaHS for24h were18.3±1.16,16.3±1.52and12.3±0.58respectively，obviouslydecreased compared with24.0±1.73of no-treatment group (P＜0.05).1.2Western blot analysis showed that the NaHS (100,200,400μmol/L) groups of theSirt1proteins expression levels were higher than the control group (P <0.05); MMP-9protein levels were lower than the blank control group (P <0.05).2.1After suppression of Sirt1expression, Wound Healing assay results show thatNaHS group’s cell migration distance132.40±15.50μm is lower than the controlgroup’s187.60±34.41μm (P<0.01); the NaHS+Sirtinol group’s cell migrationdistance223.60±31.67μm is higher than the NaHS group’s132.40±15.50μm (P<0.001); the control group’s cell migration distance187.60±34.41μm is lower thanthe Sirtinol group’268.20±35μm (P<0.01). Transwell assay results show that NaHSgroup through the Matrigel of cells13.3±0.58were lower than the control group30±1.73(P <0.01); the NaHS+Sirtinol group through the Matrigel of cells48.7±8.96were higher than the NaHS group13.3±0.58(P <0.01); the control group through theMatrigel of cells30±1.73were lower than the Sirtinol group105±9(P <0.05).2.2After suppression of Sirt1expression, Western blot showed that the NaHS group’sMMP-9protein expression levels were lower than the control group (P＜0.01); theNaHS+Sirtinol group’s MMP-9protein expression levels were higher than the NaHS group (P＜0.01); the control group’s MMP-9protein expression levels were lowerthan the Sirtinol group (P＜0.01).【Conclusion】1. H2S can inhibit the migration of hepatocellular carcinoma HepG-2cells, which maybe related to the increase Sirt1, reduced MMP-9expression.2. Sirt1mediated hydrogen sulfide were inhibited of hepatoma HepG-2cell migration,which may be related with reduced MMP-9.