The Research Of Curcumin Induced Gastric Cancer SGC-7901 Cells Apoptosis Mechanism

Objective: In this study we are going to observe curcumin inhibits human gastric cancer cells( SGC-7901)and its possible role in cell cycle,evaluate the effect of curcumin for the nf-kappa B, Livin, Caspase 3 expression. So we can provide more evidences of curcumin which could inhibit human gastric cancer cells.Discussion the possible mechanism of curcumin induced human gastric cancer SGC-7901 cells apoptosis. Method:(1)When the SGC-7901 cells in the logarithmic phase,used 1000 cells per hole added to the 96 holes in the cell culture plate,the inoculation of volume is 100μL;Under the condition of 37 ℃and 5% CO2 stick wall after 12 h, then added10,20,40,60, 80,100μmol/L different concentrations of curcumin group and DMSO solvent control group and blank control group,volume is 100μL,repeat each concentration set five hole,respectively to foster12 h, 24 h, 36 h,48h,72 h after pour culture supernatants,to join the CCK 8 fluid,by enzyme standard meter measure the energy value of each cell.(2) According to the method of CCK 8 screening in the control group and 20 、 40 、 60μmol/L curcumin concentration groups,the logarithmic phase of SGC-7901 cells,5000 cells per hole added to the 6 holes in the cell culture plate,after being cell count of 106/hole,add 2ml 1640 medium respectively,20,40,60μmol/L curcumin solution,cultivating 24 h,using flow cytometry instrument to measure each cell of the cell cycle and apoptosis rate;(3) Selection of blank control group and 20,40,60μmol/L concentrations of curcumin groups,groups of cells protein,Western Blot test groups of SGC-7901 cells nf-kappa B,Livin and Caspase-3 in gastric cancer cell apoptosis related proteins expression changes.(4)Using SPSS18.0 statistical analysis software to analyze the data,the experimental group and the control groups were compared using univariate paired samples T test was used to compare two or more data analysis of variance. Results:1.The CCK 8 testing data shows that: with the increase of curcumin drug concentration and action time,the SGC-7901 cell proliferation activity gradually reduced.Using analysis of variance each drug concentration groups of experimental data from different time cell proliferation activity,has significant statistical difference(P < 0.05).Pearson correlation analysis, according to the results of SGC-7901 cells with different concentrations of curcumin has a dose dependent(r = 0.901,P = 0.00 < 0.01), and the dependence of the time(r = 0.389,P = 0.00 < 0.01). 2.We used Flow Cytometry Method analysis of gastric cancer cell SGC-7901 cell cycle and apoptosis rate : as the concentration of curcumin was increase and time was extension,the SGC-7901 cells G1 phase cells was significantly increased,S phase cell number was decreased,shows that curcumin can block cell growth in G1 phase. 3.Western blot experiment showed that as curcumin drug concentration increased,can downregulation the NF-kappa B protein expression levels,Livin protein expression level gradually reduced at the same time, Caspase-3 protein expression level increased,and there are obvious effects of dependencies. Conclusions : 1.Curcumin inhibition the human gastric cancer SGC-7901 cells proliferation activity and has a concentration and time dependent in vitro;2.Curcumin might affect the gastric cancer cell cycle, and block in G1 phase;3.Curcumin could decreasing the NF-kappa B signaling pathway expression levels,at the same time reduce the Livin protein expression level,increasing the Caspase-3 protein expression level, and there are obvious effects of dependencies.