Study on the mechanism of procyanidin mediated anti gastric cancer SNU-1 cell line by inducing the production of reactive oxygen species / 国际肿瘤学杂志

ABSTRACT

Objective:To investigate the effect and molecular mechanism of procyanidin on the proliferation, apoptosis and reactive oxygen species (ROS) level of gastric cancer cell line SNU-1 in vitro. Methods:SNU-1 cells were divided into control group and 12.5, 50.0, 200.0 μg/ml procyanidin groups. The effect of procyanidin on the proliferation of SNU-1 cells was detected by CCK-8 assay. The apoptosis level and ROS positive rate of cells were detected by flow cytometry, and 2 mmol/L glutathione was added to SNU-1 cells added with 200.0 μg/ml procyanidin to detect the apoptosis level and ROS positive rate of cells. The expression of apoptosis-related protein in cells was detected by Western blotting.Results:The results of CCK-8 experiment showed that the proliferation activities of SNU-1 cells in the control group and the 12.5, 50.0, 200.0 μg/ml procyanidin groups were 3.69±0.30, 3.29±0.41, 0.91±0.39, 0.45±0.22 respectively, with a statistically significant difference ( F=279.84, P<0.001) . Compared with the control group, the proliferation activities of SNU-1 cells in the three procyanidin groups were significantly inhibited ( P=0.006, P<0.001, P<0.001) . The results of flow cytometry showed that the early apoptosis rates of SNU-1 cells in the control group and the 12.5, 50.0, 200.0 μg/ml procyanidin groups were (0.00±0.00) %, (0.00±0.00) %, (0.09±0.07) % and (0.45±0.22) % respectively, with a statistically significant difference ( F=7.14, P=0.003) . The 50.0 and 200.0 μg/ml procyanidin groups increased significantly compared with the control group ( P=0.003, P=0.007) . The late apoptosis rates of SNU-1 cells in the four groups were (0.00±0.00) %, (0.01±0.00) %, (6.98±0.77) % and (33.32±2.78) % respectively, with a statistically significant difference ( F=654.28, P=0.003) . The 50.0 and 200.0 μg/ml procyanidin groups increased significantly compared with the control group ( P<0.001, P<0.001) . The positive rates of ROS in SNU-1 cells in the four groups were (0.02±0.01) %, (0.10±0.05) %, (1.15±0.26) % and (1.58±0.22) % respectively, with a statistically significant difference ( F=162.24, P<0.001) . The 50.0 and 200.0 μg/ml procyanidin groups increased significantly compared with the control group ( P<0.001, P<0.001) . The positive rates of ROS in SNU-1 cells in the 200.0 μg/ml procyanidin group and the glutathione intervention group were (1.25±0.63) % and (0.13±0.02) % respectively, with a statistically significant difference ( t=5.39, P=0.001) . The early apoptosis rates of the two groups were (10.56±3.24) % and (2.09±0.24) % respectively, and the late apoptosis rates were (29.65±6.01) % and (23.63±1.52) % respectively, with statistically significant differences ( t=2.61, P=0.048; t=3.97, P=0.012) . The expressions of Bcl-2 protein in SNU-1 cells in the control group and the 12.5, 50.0, 200.0 μg/ml procyanidin groups were 1.00±0.00, 0.83±0.05, 0.60±0.14 and 0.41±0.23 respectively, with a statistically significant difference ( F=10.63, P=0.004) . The 50.0 and 200.0 μg/ml procyanidin groups decreased significantly compared with the control group ( P<0.001, P<0.001) . Conclusion:Procyanidin can inhibit proliferation and promote apoptosis of gastric cancer SNU-1 cells in vitro, which may be achieved by increasing intracellular ROS levels and reducing Bcl-2 protein expression.