阿司匹林对人胃癌细胞SGC-7901、BGC-823生长和自噬的影响
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篇名: 阿司匹林对人胃癌细胞SGC-7901、BGC-823生长和自噬的影响
TITLE:
摘要: 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2.5 μmol/L氯喹、2.5 μmol/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、5 mmol/L阿司匹林以及5 mmol/L阿司匹林单用和分别与2.5 μmol/L氯喹、2.5 μmol/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G1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,G1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G1期,其作用机制可能与激活细胞自噬有关。
ABSTRACT: 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 2.5 μmol/L chloroquine, 2.5 μmol/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 2.5 μmol/L chloroquine and 2.5 μmol/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-Ⅱ was 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.
期刊: 2019年第30卷第5期
作者: 李强,徐倩楠,廖仲恺,邓慧鸣,许荣华
AUTHORS: LI Qiang,XU Qiannan,LIAO Zhongkai,DENG Huiming,XU Ronghua
关键字: 胃癌细胞;阿司匹林;细胞自噬;细胞周期分布;存活率;凋亡率
KEYWORDS: Gastric cancer cell; Aspirin; Cell autophagy; Cell cycle distribution; Survival rate; Apoptosis rate
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