国际肿瘤学杂志

国际肿瘤学杂志 ›› 2022, Vol. 49 ›› Issue (5): 257-262.doi: 10.3760/cma.j.cn371439-20220210-00048

• 论著 •    下一篇

原花青素通过诱导活性氧产生介导抗SNU-1胃癌细胞株的作用机制研究

杨娅1, 宁晓飞2, 李炳亮3, 姚慧1, 山长平4(), 吕敏4   

  1. 1济宁医学院附属医院健康管理中心,济宁 272001
    2济宁医学院附属医院胃肠外科,济宁 272001
    3济宁医学院附属医院关节与运动医学科,济宁 272001
    4济宁医学院附属医院肿瘤科,济宁 272001
  • 收稿日期:2022-02-10 修回日期:2022-03-25 出版日期:2022-05-08 发布日期:2022-05-31
  • 通讯作者: 山长平,孙娜 E-mail:scp9933@163.com

Study on the mechanism of procyanidin mediated anti gastric cancer SNU-1 cell line by inducing the production of reactive oxygen species

Yang Ya1, Ning Xiaofei2, Li Bingliang3, Yao Hui1, Shan Changping4(), Lyu Min4   

  1. 1Health Management Center of Affiliated Hospital of Jining Medical University, Jining 272001, China
    2Department of Gastrointestinal Surgery, Affiliated Hospital of Jining Medical University, Jining 272001, China
    3Department of Joint and Sports Medicine, Affiliated Hospital of Jining Medical University, Jining 272001, China
    4Department of Oncology, Affiliated Hospital of Jining Medical University, Jining 272001, China
  • Received:2022-02-10 Revised:2022-03-25 Online:2022-05-08 Published:2022-05-31
  • Contact: Shan Changping E-mail:scp9933@163.com

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摘要：

目的 初步探究原花青素在体外对胃癌细胞株SNU-1增殖、凋亡、活性氧（ROS）水平的影响和分子机制。方法 SNU-1细胞分为对照组和12.5、50.0、200.0 μg/ml原花青素组,使用CCK-8实验检测原花青素对SNU-1细胞增殖的影响;流式细胞术检测细胞的凋亡水平和ROS阳性率,并向加入200.0 μg/ml原花青素的SNU-1细胞中加入2 mmol/L谷胱甘肽,检测细胞的凋亡水平和ROS阳性率;采用蛋白质印迹法检测细胞中凋亡相关蛋白的表达水平。结果 CCK-8实验结果显示对照组和12.5、50.0、200.0 μg/ml原花青素组的SNU-1细胞增殖活力分别为3.69±0.30、3.29±0.41、0.91±0.39、0.45±0.22,差异有统计学意义（F=279.84,P<0.001）;与对照组比较,3个原青花素组SNU-1细胞的增殖被显著抑制（P=0.006,P<0.001,P<0.001）。流式细胞术结果显示,对照组和12.5、50.0、200.0 μg/ml原花青素组的SNU-1细胞早期凋亡率分别为（0.00±0.00）%、（0.00±0.00）%、（0.09±0.07）%、（0.45±0.22）%,差异有统计学意义（F=7.14,P=0.003）;50.0和200.0 μg/ml原青花素组较对照组显著增加（P=0.003,P=0.007）。4组SNU-1细胞晚期凋亡率分别为（0.00±0.00）%、（0.01±0.00）%、（6.98±0.77）%、（33.32±2.78）%,差异有统计学意义（F=654.28,P=0.003）;50.0和200.0 μg/ml原青花素组较对照组显著增加（P<0.001,P<0.001）。4组SNU-1细胞ROS阳性率分别为（0.02±0.01）%、（0.10±0.05）%、（1.15±0.26）%、（1.58±0.22）%,差异有统计学意义（F=162.24,P<0.001）;50.0和200.0 μg/ml原青花素组较对照组显著增加（P<0.001,P<0.001）。200.0 μg/ml原花青素组和谷胱甘肽干预组SNU-1细胞的ROS阳性率分别为（1.25±0.63）%、（0.13±0.02）%,差异具有统计学意义（t=5.39,P=0.001）;2组细胞早期凋亡率分别为（10.56±3.24）%、（2.09±0.24）%,晚期凋亡率分别为（29.65±6.01）%、（23.63±1.52）%,差异均具有统计学意义（t=2.61,P=0.048;t=3.97,P=0.012）。对照组和12.5、50.0、200.0 μg/ml原花青素组SNU-1细胞Bcl-2蛋白表达水平分别为1.00±0.00、0.83±0.05、0.60±0.14、0.41±0.23,差异有统计学意义（F=10.63,P=0.004）;50.0和200.0 μg/ml原青花素组较对照组显著减少（P<0.001,P<0.001）。结论 原花青素在体外对胃癌SNU-1细胞具有抑制增殖和促进凋亡的作用,可能是通过升高细胞内的ROS水平和减少Bcl-2蛋白表达实现的。

关键词: 原花青素, 胃肿瘤, SNU-1细胞, 活性氧, 细胞凋亡

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.

Key words: Procyanidin, Gastric neoplasms, SNU-1 cells, Reactive oxygen species, Apoptosis