Study Of 1-methyl-hydantoin And Cisplatin Of Proliferation,Invasion And Metastasis In SGC-7901 Cell

Objective: The experiment is to study the mechanisms of cell proliferation, migration and invasion by 1-Methyl-Hydantoin and Cisplatin in human gastric cancer SGC-7901 cells, which provide the basis for cancer targets and clinical applications.Methods: 1. MTT assay was used to detect inhibition rate. The experiment was divided into four groups: control group(PBS), 1-Methyl1-Methyl-Hydantoin(M), DDP and 1-Methyl1-Methyl-Hydantoin combination with DDP(M+DDP). MTT was used to detect gastric cancer SGC-7901 cell proliferation inhibition rate after treating by different drugs and concentration at different time(24h, 48 h, 72h). 2. Wound-healing assays was used to observe the metastasis ability of gastric cancer SGC-7901 cell after treating by 1-methyl-hydantoin, cisplatin, and combination of the two drugs. 3. Transwell assay was used to explore invasion of gastric cancer SGC-7901 cell after treating by 1-methyl-hydantoin, cisplatin, and combination of the two drugs. 4. RT-PCR was used to detect the m RNA level of MMP9 and CD44V6 after treating by 1-methyl-hydantoin, cisplatin, and combination of the two drugs. 5. Western blotting was used to detect the protein level of MMP9 and CD44V6 after treating by 1-methyl-hydantoin, cisplatin, and combination of the two drugs.Results: 1.MTT assay results: ① The proliferation of SGC-7901 cells can be suppressed by 1-Methylhydantoin and DDP(the range of experimental concentrations is 0.5-80 umol/l). The intensity of inhibition rate dependend on concentration and time, as the concentration and the reaction time increasing the rate of inhibition was increased. ② 1-Methylhydantoin groups: compared with the control group, each group were significantly inhibited the proliferation of SGC-7901 cells(P<0.05).The rate of inhibition was best in 48 h when the concentration was 20 u mol/l. There were significant differences(p <0.05) among treatment groups. ③ DDP groups: Each group were significantly inhibited the proliferation of SGC-7901 cells(P<0.05), there were significant differences(P<0.05) at different time(24h, 48 h, 72h). And among the groups, the rate of inhibition was better in 48 h when the concentration was 1u mol/l. ④1-Methylhydantoin combination with DDP: Under the same concentration, the combination groups was stronger than the other groups on inhibiting of proliferation of SGC-7901 cells(p <0.05). 2. The scratches results showed: When SGC-7901 cells were treated for 24 h by 1-Methylhydantoin and cisplatin, injuries began to repair, and the experimental group repaired less compared with the control group,the width became smaller. After 48 h, scratches in the control group were narrowed, almost in healing,; and scratch changed smaller in the experimental group than in the control group, and the best effect was1-Methylhydantoin combination with cisplatin. The scratches results showed:the motorability of cancer cells could be weaken by treated with 1-methyl-hydantoin and cisplatin. 1-Methyl-hydantoin and cisplatin can inhibit cell migration, there was a significant difference in the experimental group and control group. 3.Transwell invasion assay results showed:compared with the control group, 1-methyl-hydantoin, cisplatin, and combination of the two drug could inhibit the invasion of gastric cancer SGC-7901 cell, the inhibition of 1-Methylhydantoin combination with cisplatin was the best. 4. Real-time quantitative(RT-PCR) results showed that MMP9, CD44V6 could express at the m RNA level in gastric cancer SGC-7901 cells. MMP9, CD44V6 expression at the m RNA level were decreased when treated with 1-methyl-hydantoin and cisplatin. 5. Western-blotting experiments showed that, compared with the control group, MMP9, CD44V6 proteins were downregulated when treated with 1-methyl-hydantoin and cisplatin. And the most obvious, which the proteins down regulated, was treated by 1-Methylhydantoin combination with cisplatin.Conclusions: 1、1-Methyl-hydantoin and DDP can inhibit the proliferation of human gastric cancer SGC7901 cells and affect cell growth. The inhibition is enhanced by the combination of the two drugs, which may have a synergistic effect. 2、1-Methyl-hydantoin and DDP can inhibit the invasion and migration of human gastric cancer SGC7901 cells. The inhibition is enhanced by the combination of the two drugs, and the expression of MMP-9 and CD44V6 decreased.