Effect And Mechanism Of Curcumin On 5-FU Induced Apoptosis In Gastric Cancer Cell Line SGC-7901

Objective1. To investigate the Curcumin combined with 5-FU in vitro culture human gastric cancer SGC-7901 cells growth inhibition; understand certain doses of Curcumin and low dose of 5-FU combined with on SGC-7901 cell growth inhibition can achieve the effect of high dose 5-FU alone.2. To explore the Curcumin combined with 5-FU in vitro culture in human gastric cancer cell line SGC-7901 cell apoptosis inducing effect; understand certain doses of Curcumin and low dose of 5-FU combined with induced apoptosis of SGC-7901 cells can reach the effect of high dose 5-FU alone.3. The effect of Curcumin combined with 5-FU in vitro cultured human gastric cancer SGC-7901 cell cycle effect; understanding of Curcumin and low dose of 5-FU combined with induced SGC-7901 cell cycle effect can reach the effect of high dose 5-FU alone.4. To explore the mechanism of Curcumin combined with 5-FU to induce the apoptosis of gastric cancer SGC-7901 cells, and to provide experimentalevidence for clinical application of Curcumin and 5-FU in the treatment of gastric cancer.Method1. Based on the enzyme mark instrument was measured by MTT assay of Curcumin(20 μ mol / L) and 5-FU(10, 20, 40 μ mol / L) used alone or in combination on the in vitro culture of human gastric cancer SGC-7901 cell growth and proliferation.2. Using PI single staining flow cytometry and AV-FITC/PI double staining flow cytometry analysis of Curcumin and its combination with SGC-7901 on the apoptosis and proliferation cycle of human gastric cancer cell line 5-FU.3. The effect of Curcumin and its combination with 5-FU on the activity of Caspase-3 and Caspase-8 in gastric cancer cell line SGC-7901 was detected by ELISA.4. To detect the expression of Bcl-2 and Bax in human gastric cancer SGC-7901 cell apoptosis related protein and 5-FU by using blot Western method.Result1. Curcumin combined with 5-FU on the proliferation of SGC-7901 cell inhibition: compared with the control group, the experimental group can significantly inhibit the proliferation of SGC-7901 cells(*P<0.05); Curcumin combined with low dose of 5-FU inhibition was higher than that in the dose of5-FU alone(P < 0.05); combined with Curcumin dose of 5-FU inhibition ratehigher than that of high dose of 5-FU alone(P < 0.05).2. Curcumin combined with 5-FU on the apoptosis of SGC-7901 cells influence: the apoptosis of SGC-7901 cells in each experimental group rate were significantly higher than those of the control group(P < 0.05); Curcumin combined with low dose of 5-FU group, cell apoptosis rate was 39.3%, which was significantly higher than that in the dose of 5-FU alone group of 26.4%(P< 0.05); combined with Curcumin dose 5-FU group cell apoptosis rate was43.9%, significantly higher than that of high dose 5-FU alone group(31.9%)(P < 0.05).3. Curcumin combined with 5-FU on proliferation of SGC-7901 cell cycle effects: compared with the control group, Curcumin and 5-FU can cause SGC-7901 cells in S phase(P < 0.05) block; Curcumin combined with low dose of 5-FU group and middle dose of 5-FU alone group compared to, in the proportion of S phase cells increased from 41.2% to 42%(P > 0.05);Curcumin combined with 5-FU dose group and high dose 5-FU alone group compared in the proportion of S phase cells by 45.6% rose to 53.8%(P <0.05).4. The influence of Curcumin combined with 5-FU on SGC-7901 cell apoptosis protease Caspase-3 and Caspase-8 activity: compared with the control group, 5-FU group and Curcumin could induce significant increase of SGC-7901 cells in Caspase-3 and Caspase-8 relative activity; Curcumin combined with low dose of 5-FU group and 5-FU group compared with singledose of Caspase-3, the relative percentage of activity by 0.14 times(1.14 +0.08) increased to 0.18 times(1.18 + 0.09) Caspase-8, the relative percentage of activity by 0.10 times(1.10 + 0.09) increased to 0.13 times(1.13 + 0.07)(*P; <0.05) of Curcumin combined with medium dose 5-FU group and high dose group compared with single 5-FU, Caspase-3, Caspase-8 relative activity respectively by 0.35 times the original(1.35 + 0.03), 0.19 times(1.19 + 0.01)increased to 0.4 times(1.40 + 0.03), 0.28 times(1.28 + 0.09)(*P <0.05).5. The influence of Curcumin combined with 5-FU on SGC-7901 cell apoptosis related protein Bcl-2 and Bax expression: compared with the control group, the expression of Bax protein in experimental group increased significantly(*P and <0.05); Curcumin combined with low dose of 5-FU group and 5-FU group compared with single dose, the expression of Bax protein significantly increased(relative value respectively. 2.56 + 0.09; 1.33 +0.01), while the expression of Bcl-2 was significantly decreased(relative values were 0.76 + 0.04; 0.88 + 0.05); Curcumin combined with medium dose5-FU group and high dose group compared with single 5-FU, Bax protein expression increased significantly(relative values were 3.27 + 0.06; 2.19 +0.05), and the expression of Bcl-2 protein was significantly decreased(relative values were 0.33 + 0.01; 0.53 + 0.02)(*P <0.05).Conclusion1. Effect of Curcumin combined with low dose 5-FU on growth inhibition and apoptosis of gastric cancer SGC-7901 cells cultured in vitro can achieve highdose of 5-FU;2. The enhancement effect of Curcumin combined with low dose 5-FU on the relative activity of Caspase-3 and Caspase-8 in gastric cancer SGC-7901 cells was significantly higher than that of high dose 5-FU alone;3. Curcumin combined with low dose 5-FU and high dose 5-FU alone in vitro culture of gastric cancer SGC-7901 cells Bax protein expression significantly increased, Bcl-2 protein expression was significantly reduced.