| Objective:To study the influence of the glelitsia sinensis lam concentration on the proliferation,the inhabiting on some cancerous gene and telomerase activity of human hepatocarcinoma cell.Methods:Four groups in random:comparison group,3 groups of the glelitsia sinensis lam concentration(0.05mg/mL,0.1mg/mL,0.2mg/mL),to record the influence of the proliferation,the inhabit on some cancerous gene and telomerase activity of human hepatocarcinoma cell bel-7402.Experiment 1:Select the parts of glelitsia sinensis lam that have initial inhabit on proliferation of human hepatocarcinoma cancerous cells.Divide 18 male Wistar rats into six groups and assign glelitsia sinensis lam concentration to the six roups.Use MTT to detect the influence of drug serum on each group on the proliferation of human hepatocarcinoma cell.Experiment 2:Further purify the parts of glelitsia sinensis lam that have initial inhabit on proliferation of human hepatocarcinoma cells.Three groups in random:comparison group,group of glelitsia sinensis lam concentration A,group of glelitsia sinensis lam concentration B,to record,by MTT,the influence of the proliferation of human hepatocarcinoma cells.Experiment 3:To observe the influence of glelitsia sinensis lam on lifecycle,apoptosis-related genes and telomerase of human hepatocarcinoma cells.Four groups in random:comparison group,3groups of the glelitsia sinensis lam concentration(0.05mg/mL,0.1mg/mL,0.2mg/mL),to record,by MTT,the influence of the proliferation of human hepatocarcinoma cells. Experiment 4:Observe the influence of glelitsia sinensis lam on lifecycle of human hepatocarcinoma cells.Four groups in random: comparison group,3 groups of the glelitsia sinensis lam concentration (0.05mg/mL,0.1mg/mL,0.2mg/mL).Choose 488nm excitation light on the flow cytometry to detect the DNA quantity of each group to calculate the apoptosis rate of human hepatocarcinoma cells.Experiment 5:To observe the influence of glelitsia sinensis lam concentration on apoptosis-related genes of human hepatocarcinoma cells.Four groups in random:comparison group,3 groups of the glelitsia sinensis lam concentration(0.05mg/mL,0.1mg/mL,0.2mg/mL),to detect, by immunohistochemical method,the influence on the genes(bax,bcl-2, p53)of human hepatocarcinoma cells.Experiment 6:To observe the influence of glelitsia sinensis lam concentration on telomersae of human hepatocarcinoma cells.Four groups in random:comparison group,3 groups of the glelitsia sinensis lam concentration(0.05mg/mL,0.1mg/mL,0.2mg/mL).The detection of telomerase activity is implemented according to the instruction in the telomerase testing kit.Experimental data checked by t test,spss10.0 statistical software used for statistical analysis.Results:Experiment 1:MTT experimental results shows that glelitsia sinensis lam ethyl acetate concentration has significant inhabit on proliferation of human hepatocarcinoma cell bel-7402(P＜0.01).Oil acid solution,ether solution,butanol solution,and water solution of glelitsia sinensis lam concentration have no statistically significance on the proliferation of bel-7402(P＞0.05).Experiment 2:MTT experimental results shows that glelitsia sinensis lam ethyl acetate further purified concentration B has significant inhabit on proliferation of human hepatocarcinoma cell bel-7402(P＜0.01).Concentration B has no statistically significance on the proliferation of bel-7402(P＞0.05).Experiment 3:MTT experimental results show that high and middle concentration groups of glelitsia sinensis lam have significant inhabit on proliferation of human hepatocarcinoma cell bel-7402(P＜0.05).Among which,high concentration group has extreme statistically significance on the proliferation of bel-7402(P＜0.01).Experiment 4:MTT experimental results show that after 72 hours,G0 G1 period of bel-7402 cells increases while S period decreases in the low,middle and high concentration groups of glelitsia sinensis lam.Compared with the comparison group,the difference is significant P＜0.01).Apoptosis peak occurs before G0 - G1.Experiment 5:The experimental results showed that the drug serum bcl-2 expression color has no significant difference compared with the comparison group(p＞0.05).Bax and p53 expression color of low,middle and high concentration serum groups are deepen compared with the comparison group,with statistical significance(p＜0.05,or P＜0.01).Results also show that influence on the bax and p53 protein expression has no obvious dependency on the consistency.Experiment 6: Compared with the comparison group,value decreased in low,middle and high concentration of serum group and comparison group A(p＜0.05,or p＜0.01),indicating that low,medium and high serum groups of glelitsia sinensis lam and the comparison group can inhibit the activity of telomerase of cell bel-7402.Among which,high concentrations of serum group has very significant inhabit on telomerase activity(p＜0.01).Conclusions:The concentration of glelitsia sinensis lam can inhabit the proliferation,accelerate the apoptotsis of bel-7402 cell,inhabit the telomerase activity of cell bel-7402.Inhabit and control cancer gene inactivation and activation of tumor suppressor genes,prevent the unlimited reproduction of tumor cell.Compared with the comparison group, high concentration group has more significance on the proliferation of tumor cells.Initially,the results show that the possible mechanisms of theglelitsia sinensis lam' s inhabit on the tumor cells are as follows:①inhibit activation of telomerase,to stop the unlimited reproduction of DNA;②enhance the activity of tumor suppressor gene,and promote cancer cell apoptosis;③as a possible Kinds of information material, directly involved in apoptosis,inhibition of tumor cell DNA unlimited reproduction.Further analysis is expected.
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