Effects Of Anti-proliferation And Induced Apoptosis By 1-methylhydantoin And 5-Fu On Gastric Cancer SGC-7901 Cells

Objective:To analysis the inhibit and induced apoptosis effects of 1-Methylhydantoin or 5-fluorouracil separate and the combination with the both in human gastric cancer cell line SGC-7901. Methods:Experimental subject: The human gastric cancer cell line SGC-7901 which were cultured in vitro. Groups: The experiment was divided into five groups: control group(PBS), 1-Methylhydantoin(5 mg·L-1, 10 mg·L-1, 20 mg·L-1, 50 mg·L-1, 100 mg·L-1), 5-fluorouracil(5 mg·L-1, 10 mg·L-1, 20 mg·L-1, 50 mg·L-1, 100 mg·L-1) and 1-Methylhydantoin combination with 5-fluorouracil(M 5 mg·L-1+Fu 5 mg·L-1, M 10 mg·L-1+Fu 10 mg·L-1, M 20 mg·L-1+Fu 20 mg·L-1, M 50 mg·L-1+Fu 50 mg·L-1, M 100 mg·L-1+Fu 100 mg·L-1).MTT assay was used to detect inhibition rate; FCM detected apoptosis and cell cycle; Western Blot detected the protein expression levels of ERK, P53, FAS, FASL. Results:1 MTT assay results:① 1-Methylhydantoin and 5-Fluorouracil could suppress proliferation of SGC-7901 cells within the range of experimental concentrations(5 ~ 100 mg·L-1), There are statistically significant differences contrast the control group(p<0.05). The intensity of inhibition rate dependend on concentration and time, as the concentration increases the rate of inhibition increased while the reaction time is increased to increase the rate of inhibition.② 1-Methylhydantoin groups: 24 h later, compared with the control group, each group were significantly inhibited the proliferation of SGC-7901 cells(p<0.05).③ 5-Fluorouracil groups: Each group were significantly inhibited the proliferation of SGC-7901 cells(p<0.05), and among the groups, there were significant differences(p<0.05) at the same time.④ Under the same concentration, the effect of 1-Methyl-hydantoin inferior to 5-fluorouracil. After 24 h, there is no significant difference between 20 mg·L-1 5-fluorouracil and 50 mg·L-1 and 100 mg·L-11-Methyl hydantoin on inhibiting proliferation of SGC-7901 cells(p>0.05). After 48 h, the difference was not significant between 50 mg·L-1 1Methylhydantoin and 10 mg·L-1 5-fluorouracil(p>0.05). After 72 h, there was no significant between 50 mg·L-1 1-Methylhydantoin and 10 mg·L-1 5-fluorouracil while also here was no significant between 100 mg·L-1 1-Methylhydantoin and 20 mg·L-1 5-fluorouracil(p>0.05).⑤1-Methylhydantoin combination with 5-fluorouracil: After 24 h, when the concentration of combination was 50 mg·L-1+50 mg·L-1, the inhibitory effect was higher than M(100 mg·L-1), meanwhile, it was significant lower than 5-Fu(100 mg·L-1)(p<0.05). After 48 h and 72 h, the effect of proliferation inhibitory were equal at the combination(50 mg·L-1+50mg·L-1) and the 5-fluorouracil(100 mg·L-1)(p>0.05), however, the two treatments were significant higher than M 100 mg·L-1)(p<0.05).2 FCM assay results: Compared with control group, there were significant differences of apoptosis rate and cell cycle of SGC-7901 cells in each treatment group of 1-Methylhydantoin, 5-Fu and 1-Methylhydantoin+5-Fu(p<0.05). Compared with 5-Fu groups and 1-Methylhydantoin groups, there were significant differences of apoptosis rate and cell cycle of SGC-7901 cells in combinaton groups(p<0.05).3 Western Blot detected results: Compared with the control group, the 1-Methyl-hydantoin treated SGC-7901 cells, extracellular regulated protein kinases(ERK), tumor suppressor gene(P53),death repeptor Fas and its ligand Fas L were no difference. The 5-fluorouracil treated SGC-7901 cells, there no difference in the expression of P53, but, the expression of ERK, Fas,and Fas L was significantly upregulated. Conclusions1.1-Methylhydantoin could inhibit proliferation and induce apoptosis of gastric cancer SGC-7901 cells in vitro.2.1-Methylhydantoin gastric cancer SGC-7901 cells in vitro in a dose and time dependent, with increasing concentration and the role of prolonged, the inhibitory effect increased.3.Under the same concentration, 1-Methylhydantoin were weaker than 5-fluorouracil on the inhibitory effect of gastric cancer SGC-7901 cells.4.1-Methylhydantoin and 5-fluorouracil combined to inhibit proliferation of gastric cancer SGC-7901 cells may have additive effect.