| [Objectives]Cancer damage human health seriously, and the incidence of it is increasing every year. After entering into the 21century, tumor researchers are considering the new ideas, new ways and new methods to cure tumor. Using the traditional Chinese medicine to cure tumor is becoming one of the most important ways. The traditional Chinese medicine is the valued fortune of our country. There are rich natural medicine materials in our country, and tumor researchers attach more importance to the effect of these medicine materials. We studied the effect of the fructus amomi rotundus acting on SGC-7901 cells. We observed the changes of the inhibition ratio, the changes of the apoptosis ratio, and the changes of the micromechanism of the cells after acting on SGC-7901 cells by using different groups of fructus amomi rotundus. At the same time, we detected the expression of bcl-2 of SGC-7901. Based on the results of the research we want to studied the mechanism of it furthermore and put it into application.[Methods]We used the eugonic SGC-7901 cells. After being digested by trypsin, SGC-7901 cells were seeded in 96-well plates(1-2×106/ml). Employed different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml) when SGC-7901 cells subsided and grew along the bottom of 96-well plate. At the same time, we set up reagent control group and tumor cell control group. After 24,48 and 72 hours in cultivate device full of 5%CO2, measured the OD value(570nm). Then calculated the inhibition ratio of different groups.After 24, 48 hours when different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml) acted on SGC-7901 cells, collected cells after being digested by using trypsin. Then washed by PBS solution, dissolved the cells in PBS solution in order to be the 106/ml concentration. Mixed in proper acridine orange and ethidium bromide solution, watched the changes of the micromechanism of the apoptosis cells with fluoroscope. Calculated the apoptosis rate of different groups.After 24, 48 hours when different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml) acted on SGC-7901 cells, collected cells after being digested by using trypsin. Then washed by PBS solution, dissolved the cells in 200ul combine buffer solution, Mixed in proper Annexin-Ⅴ-FITC and PI, placed it in room temperature after 15 minutes, detected the apoptosis rate by FCM.After 48 hours when different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml) acted on SGC-7901 cells, collected the cells after being digested by using trypsin. Then embed the cell with wax. Dyed slice by SP after making cell wax slice. Observed the expression of bcl-2 with image-analysing system of different groups.[Results]After different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml) acted on SGC-7901 cells, MTT assay founded that the inhibition ratio are 8.03±2.39,25.18±1.53,30.99±1.96,40.40±1.94 after 24 hours; the inhibition ratio are 23.76±1.58,32.37±1.75,41.53±1.97,58.78±1.97 after 48 hours; the inhibition ratio are 29.93±1.79,41.32±1.97,49.24±1.57,64.65±1.33 after 72 hours. There are significant decrease between the control group and the experimental groups( P<0.05).Through observing the effects of the different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml) acted on SGC-7901 cells by AO-EB fluorescent staining method, we found the apoptosis rate are 2.50±0.50,11.33±1.04,18.50±1.32,24.00±0.50,33.83±2.08 after 24 hours; the apoptosis rate are 4.67±1.04,19.00±1.73,26.83±2.02,33.50±2.29,41.67±0.29 after 48 hours. There are significant decrease between the control group and the experimental group s ( P<0.05). After different concentration of fructus amomi rotundus (0.2mg/ml,0.4mg/ml, 0.8mg/ml,1.0mg/ml) acted on SGC-7901 cells, we detected the apoptosis rate by FCM. The early apoptosis rate are 1.43±0.15,9.87±0.21,16.43±0.85,20.80±1.90,26.77±0.35 after 24 hours; The early apoptosis rate are 2.07±0.25,16.13±1.07,20.47±0.87,23.83±0.91,35.37±1.36 after 48 hours. There are significant decrease between the control group and the experimental groups ( P<0.05).Observed the expression of bcl-2 with image-analysing system in different experimental groups. We found the average light consistency are 0.419±0.016,0.374±0.010,0.286±0.011,0.183±0.014 in the control group,0.4mg/ml,0.8mg/ml,1.0mg/ml group.There are significant decrease between the control group and the experimental groups( P<0.05).Conclusion:1,0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml fructus amomi rotundus group can inhibit the growth of SGC-7901 cells.2,0.2mg/ml,0.4mg/ml,0.8mg/ml,1.0mg/ml fructus amomi rotundus group can induce apoptosis of SGC-7901 cells.3,Some concentration of fructus amomi rotundus can lower the expression of bcl-2, it maybe promotes SGC-7901 cells apoptosis. |
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