阿司匹林对人胃癌细胞SGC-7901、BGC-823生长和自噬的影响

目的:研究阿司匹林对人胃癌细胞SGC-7901、BGC-823生长和自噬的影响。方法:以SGC-7901、BGC-823细胞为研究对象,磷酸盐缓冲液(PBS)为阴性对照组,作用时间为48 h,采用MTT法检测1、2、4、6、8、10 mmol/L阿司匹林以及5 mmol/L阿司匹林单用和分别与2.5μmol/L氯喹、2.5μmol/L 3-甲基腺嘌呤(3-MA)联用对胃癌细胞存活率的影响;采用流式细胞术检测2、5mmol/L阿司匹林以及5 mmol/L阿司匹林单用和分别与2.5μmol/L氯喹、2.5μmol/L 3-MA联用对胃癌细胞凋亡率和细胞周期分布的影响;采用Hoechst33258染色观察5 mmol/L阿司匹林对胃癌细胞核形态学的影响;采用Transwell小室试验检测5 mmol/L阿司匹林对胃癌细胞迁移的影响;采用激光共聚焦扫描显微镜观察5 mmol/L阿司匹林对胃癌细胞内自噬体形成的影响;采用Western blot法检测2、5 mmol/L阿司匹林对胃癌细胞内自噬标志物LC3-Ⅱ蛋白表达的影响。结果:与阴性对照组比较,阿司匹林能明显抑制SGC-7901、BGC-823细胞的存活率,且呈浓度依赖性,但对SGC-7901、BGC-823细胞的凋亡率没有明显影响,能够诱导细胞周期阻滞在G_1期。与阿司匹林单用组比较,阿司匹林+氯喹和阿司匹林+3-MA作用后SGC-7901、BGC-823细胞存活率明显升高,G_1期细胞分布率明显降低,差异均有统计学意义(P<0.05或P<0.01)。与阴性对照组比较,阿司匹林作用后SGC-7901、BGC-823细胞均未见明显的DNA裂解片段、凋亡小体以及碎块状的浓密亮蓝色,迁移数量明显减少(P<0.001),自噬体明显增加,LC3-Ⅱ蛋白表达明显增强(P<0.05)。结论:阿司匹林能够显著抑制胃癌细胞SGC-7901、BGC-823的生长,使其细胞周期停滞在G_1期,其作用机制可能与激活细胞自噬有关。

Effects of Aspirin on the Growth and Autophagy of Human Gastric Cancer Cells SGC-7901 and BGC-823

OBJECTIVE:To study the effects of aspirin on the growth and autoghagy of human gastric cancer cells SGC-7901 and BGC-823.METHODS:SGC-7901 and BGC-823 cells were selected as research objects,with phosphate buffer(PBS)as negative control treated for 48 h,MTT assay was used to detect the effects of 1,2,4,6,8,10 mmol/L aspirin,5 mmol/L aspirin alone or combined with 2.5μmol/L chloroquine,2.5μmol/L 3-methyladenine(3-MA)on survival rate of gastric cancer cells.Flow cytometry was used to detect the effects of 2 and 5 mmol/L aspirin,5 mmol/L aspirin alone or combined with 2.5μmol/L chloroquine and 2.5μmol/L 3-MA on the apoptosis rate and cell cycle distribution of gastric cancer cells.Hoechst33258 staining was used to observe the effects of 5 mmol/L aspirin on morphology of gastric cancer cell nucleus;Transwell chamber test was adopted to detect the effects of 5 mmol/L aspirin on the migration of gastric cancer cell.Laser confocal scanning microscopy was used to observe the effects of 5 mmol/L aspirin on autophagy formation in gastric cancer cells.Western blot method was used to detect the effects of 2 and 5 mmol/L aspirin on the protein expression of autophagy markers LC3-Ⅱin gastric cancer cells.RESULTS:Compared with negative control group,aspirin could inhibit the survival rates of SGC-7901 and BGC-823 cells in dose-dependent manner,but had no significant effects on apoptosis rate of SGC-7901 and BGC-823 cells;SGC-7901 and BGC-823 cells were blocked in G1 phase.Compared with aspirin alone group,the survival rates of SGC-7901 and BGC-823 were increased significantly after treated with aspirin+chloroquine and aspirin+3-MA,while the distribution rate of SGC-7901 and BGC-823 cells at G1 phase were decreased significantly,with statistical significance(P<0.05 or P<0.01).Compared with negative control group,there were no obvious DNA fragmentation fragments,apoptotic bodies and fragments of dense bright blue,while the number of migration cells were decreased significantly in SGC-7901 and BGC-823 cells after treated with aspirin(P<0.001);the number of autophagosome was increased significantly and the protein expression of LC3-Ⅱwas enhanced significantly(P<0.05).CONCLUSIONS:Aspirin can significantly inhibit the growth of SGC-7901 and BGC-823 cells,and arrest cell cycle in G1 phase,the mechanism of which may be associated with the activation of autophagy.