Study On The Effect Of Heparanase Gene Over-expression On The Proliferation,Apoptosis And Migration Of Gallbladder Carcinama Cells And Its Mechanism

Objective:To investigate the effect of Heparanase gene over-expression on the proliferation,apoptosis and migration of gallbladder cancer cells,and to preliminary study its related molecular mechanism.Methods:1.CCK-8 assay and colony formation experiment were used to detect the proliferation of GBC-SD and GBD-SD cells.2.Hoechst 33324 staining assay and flow cytometry were used to detect the apoptosis of GBC-SD and GBD-SD cells.3.Scratch wound experiment was used to detect the migration ability of GBC-SD and GBD-SD cells.4.Western blot was used to detect the relative protein expression of Syndecan-1,FGF-2,E-cadherin,?-catenin,N-cadherin and Vimentin in GBC-SD and GBD-SD cells.5.Cellular immunofluorescence staining was used to detect E-cadherin and Vimentin expression and cell localization in GBC-SD and GBD-SD cells.Results:1.The results of CCK-8 experiment showed that the cell proliferation ability of GBD-SD group was significantly higher than that of GBC-SD group(P<0.05);The results of colony formation experiment showed that the cell colony forming ability of GBD-SD group increased significantly higher compared with GBC-SD group(P<0.05).2.Hoechst 33324 staining assay results showed that the rate of cell apoptosis in GBD-SD group was significantly lower than that in GBC-SD group(P<0.05);Flow cytometry results showed that the percentage of cell apoptosis in GBD-SD group was significantly reduced compared with GBC-SD group(P<0.05).3.The results of scratch wound experiment showed that the cell migration ability of GBD-SD group was significantly higher than that of GBC-SD group(P<0.05).4.Western blot results showed that compared with GBC-SD group,the relative protein expression of FGF-2?N-cadherin and Vimentin in GBD-SD group increased significantly higner(P<0.05),while the relative expression of Syndecan-1?E-cadherin and ?-catenin protein decreased significantly lower(P <0.05).5.The results of cell immunofluorescence staining showed that compared with the GBC-SD group,the staining intensity of E-cadherin in the GBD-SD group was weakened,while the staining in Vimentin was significantly enhanced.Conclusion:1.Heparanase gene over-expression up-regulates cell proliferation and migration ability,and down-regulates the level of cell apoptosis in gallbladder cancer cells.2.The mechanism which Heparanase regulates the biological behavior of gallbladder carcinoma cells may be related to the regulation of EMT through FGF-2.