Study Of Granzyme B Gene Combined Curcumin Affect Huh7Hepatoma Cell

Objective:To amplified and identified granzyme B-LV183lentiviral vecto.Methods:Granzyme B-LV183transfection of Huh7hepatoma cells,caclculate the transfection efficiency, and validated by RT-PCR. The PCR amplification products was verified by sequencing.Results:The amplified lentiviral vector OD260/280=1.87, electrophoresis results was clear. Granzyme B-LV183transfected into Huh7hepatoma cells,the transfection efficiency was about70%, by RT-PCR detection of gene expression. PCR products were sequenced and the results are correct.Conclusions:1.We successfully transfected Granzyme B-LV183into Huh7hepatoma cells. Part2Study of granzyme B combined Curcumin affect Huh7hepatoma cellObjective:To study the impact of transfected GranzymeB gene combines curcumin on Huh7hepatoma cells.Methods:Based on part one, The experiment is divided into five groups: normal control group (KB), the curcumin group (GH), GranzymeB transfection group (GrB), GranzymeB curcumin group (GG), empty plasmid (KZL).40μmol/L concentrations of curcumin solution in the curcumin group and GranzymeB curcumin group (6,24,96orifice in the GG and GH group were added of500μl,200μl,20μl has been configured40μmol/L concentration of turmeric prime solution).Detect apoptosis by flow cytometry, detect cell proliferation by CCK-8.Results:The concentration of ALT,AST,AFP in each curcumin dosing group, P<0.05, has significant difference; Compared in each pairwised group, the GG group and GrB group, the curcumin group, blank group, the empty plasmid, P<0.05,has significant difference; The GrB group compared with the curcumin group, P>0.05, has no significant difference. Apoptosis rate of each group, P <0.05, has significant difference; GG group compared with the the GrB group and curcumin group, blank group, empty vector group, P <0.05, has significant difference; the GrB group compared with the curcumin group, P>0.05, has no significant difference; Cell proliferation test of each group, P<0.05, has significant difference; Compared the GG group to GranzymeB group, blank group and curcumin group, the empty plasmid group, P<0.05, has significant difference; GranzymeB group and curcumin group P>0.05has no significant difference.Conclusions:1.GranzymeB joint effect of curcumin on liver cancer cells to inhibit the proliferation of hepatoma cells,and promote apoptosis of hepatoma cells.