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  

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

作　　者：杨娅[1] 宁晓飞[2] 李炳亮[3] 姚慧 山长平[4] 吕敏 Yang Ya;Ning Xiaofei;Li Bingliang;Yao Hui;Shan Changping;Lyu Min(Health Management Center of Affiliated Hospital of Jining Medical University,Jining 272001,China;Department of Gastrointestinal Surgery,Affiliated Hospital of Jining Medical University,Jining 272001,China;Department of Joint and Sports Medicine,Affiliated Hospital of Jining Medical University,Jining 272001,China;Department of Oncology,Affiliated Hospital of Jining Medical University,Jining 272001,China)

机构地区：[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

出　　处：《国际肿瘤学杂志》2022年第5期257-262,共6页Journal of International Oncology

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、200.0μg/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200.0μg/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、200.0μg/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、200.0μg/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200.0μg/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200.0μg/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200.0μg/ml原青花素组较对照组显著增加(P<0.001,P<0.001)。200.0μg/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、200.0μg/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�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 12.5,50.0,200.0μg/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 200.0μg/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 12.5,50.0,200.0μg/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 12.5,50.0,200.0μg/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 200.0μg/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 200.0μg/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

关 键 词：原花青素 胃肿瘤 SNU-1细胞 活性氧 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]