Effects Of Diallyl Disulfide On Migration And Invasion In Gastric Cancer MGC803 Cells Of Overexpression Of RhoGDI2

Objective: Our former studies have proved that diallyl disulfide(DADS) could suppress migration and invasion of MGC803 cells by down-regulation of LIMK1 expression via blocking Rac1-Pak1/Rock signalway.While proteomics found that DADS could cut the expression of Rho GDI2.In this study,we will investigate the effect of DADS on biological behaviour in MGC803 cell of overexpressed Rho GDI2 by establishing a stable MGC803 cells of overexpressed RhoGDI2.Methods: We transfected the overexpression of Rho GDI2 gene to MGC803, finally we established Rho GDI2 overexpression MGC803 cells.We used Western blotting assay to detect the expressions level of Rho GDI2 protein. We used wound healing and transwell invasion chamber assays to disscuss the effect of DADS on migration and invasion in Rho GDI2 overexpression MGC803 cells. We used nude mice xenograft experiments to disscuss the effect of DADS on tumorigenicity in Rho GDI2 overexpression MGC803 cells.Results 1. Establish a stable MGC803 cells line of overexpressed Rho GDI2. We could find green fluorescence in two groups of transfection cells, Rho GDI2/MGC803 and p IRES2-EGFP/MGC803. Western Blot proved that the expression level of Rho GDI2 protein in Rho GDI2/MGC803 were higher than that ofMGC803 and p IRES2-EGFP/MGC803. It showed that we established a stable MGC803 cells line of overexpressed Rho GDI2. 2. The effect of DADS on migration in MGC803 cell of overexpressed Rho GDI2. Wound healing demonstrated that the nick distance of Rho GDI2/MGC803 were lower than that of MGC803 and p IRES2-EGFP/MGC803(P<0.05).It showed that overexpression of Rho GDI2 could promote the migration of MGC803. The nick distance of cells dealed with DADS were significantly higher than that of of cells without DADS(P<0.05).It shows that DADS can reduce the migration of MGC803 and Rho GDI2/MGC803. 3. Effect of DADS on invasion in MGC803 cell of overexpressed Rho GDI2. Transwell invasion chamber assays demonstrated that the numbers of cells through the cell membrane of Rho GDI2/MGC803 were higher than that of MGC803 and p IRES2-EGFP/MGC803(P<0.05).It showed that overexpression of Rho GDI2 could promote the invasion of MGC803. The numbers of cells through the cell membrane of cells dealed with DADS were significantly lower than that of cells without DADS(P<0.05).It showed that DADS can reduce the invasion of MGC803 and Rho GDI2/MGC803. 4.Effect of DADS on tumorigenicity in MGC803 cell of overexpressed Rho GDI2. Nude mice xenograft experiments found that the growth of xenograft tumor of MGC803 groups were lower than that of Rho GDI2/MGC803 cells groups(P<0.05). It proved that the high expression of Rho GDI2 promote the growth of nude mice xenograft tumor.The growth rate of xenograft tumor of cells dealed with DADS were significantly below to that of the cells without DADS(P<0.05).It proved that DADS significantly suppressed tumorigenicity of MGC803 and Rho GDI2/MGC803. Immunohistochemistry of xenograft show that the expression of E-cadherin dealed with DADS is higher than that of without DADS. While the expression of Vimentin、Ki-67 and CD34 dealed with DADS is lower than that of without DADS. The expression of E-cadherin in Rho GDI2/MGC803 is lower than that of MGC803. While the expression of Vimentin、Ki-67 and CD34 in Rho GDI2/MGC803 is higherr than that of MGC803.Conclusion 1. DADS inhibit migration invasion and the growth rate of nude mice xenograft tumor in Rho GDI2 overexpression MGC803 cells. 2. Rho GDI2 overexpression can increase the migration invasion and growth of nude mice xenograft tumor in MGC803 cells.