摘要
目的:探讨芦荟大黄素(AE)对胃癌SGC-7901细胞凋亡、自噬及p53蛋白(p53)/磷酸腺苷蛋白激酶(AMPK)/雷帕霉素靶蛋白(m TOR)信号通路的影响。方法:实验分为对照组(AE 0μmol·L^-1)、AE低(10μmol·L^-1)、中(30μmol·L^-1)、高(50μmol·L^-1)剂量组和自噬阻断剂3-甲基腺嘌呤(3-MA)+AE组(20μmol·L^-1+50μmol·L^-1)。CCK-8法检测各组细胞增殖抑制率,采用透射电镜及单丹磺酰尸胺(MDC)染色观察各组细胞自噬体及自噬溶酶体的变化,采用末端脱氧核苷酸转移酶介导的d UTP缺口末端标记测定(TUNEL)法观察各组细胞凋亡率变化,Western blot法检测各组细胞p53、AMPK、m TOR信号通路蛋白及自噬标致蛋白Beclin-1蛋白(Beclin-1)、微管相关轻链蛋白Ⅰ(LC3Ⅰ)及微管相关轻链蛋白Ⅱ(LC3Ⅱ)表达情况。结果:与对照组相比,AE各剂量组及3-MA+AE组细胞的增殖抑制率、凋亡指数(AI)显著升高(P<0.05),且呈剂量依赖性;AE中、高剂量组的增殖抑制率和AI与3-MA+AE组相比差异有统计学意义(P<0.05)。与对照组相比,AE各剂量组细胞自噬体、自噬空泡增多,溶酶体积分光密度(IOD)/所选区域面积(Area)值及p53、AMPK、Beclin-1表达显著升高,m TOR表达及LC3Ⅰ/LC3Ⅱ比值显著降低(P<0.05),且呈剂量依赖性;3-MA+AE组细胞自噬体、自噬空包等超微结构减少,IOD/Area值及p53、AMPK及Beclin-1表达显著降低,m TOR表达及LC3Ⅰ/LC3Ⅱ比值显著升高(P<0.05)。结论:AE可通过提高胃癌SGC-7901细胞自噬水平,诱导SGC-7901细胞凋亡,其机制可能与促进p53/AMPK/m TOR信号通路表达有关。
Objective: To investigate the effects of aloe emodin( AE) on apoptosis,autophagy and p53/AMPK/m TOR signaling pathway in gastric cancer SGC-7901 cells. Methods: The experiment was divided into the control group( AE 0 μmol·L^-1),AE low( 10 μmol·L^-1),medium( 30 μmol·L^-1) and high( 50 μmol·L^-1) dose groups and autophagy blocker 3-methyladenine( 3-MA)+ AE group( 20 μmol·L^-1+ 50 μmol·L^-1). CCk-8 method was used to detect the inhibition rate of cell proliferation,the changes of autophages and autophagic lysosomes were observed by transmission electron microscopy and MDC staining,TUNEL method was used to observe the changes of apoptotic rate,and the expressions of p53/AMPK/m TOR signaling pathway protein and autophagy marker protein Beclin-1,LC3 I and LC3 II were detected by Western blot. Results: Compared with those in the control group,the proliferation inhibition rate and the apoptotic index( AI) of AE group and 3-MA + AE group were significantly increased( P < 0. 05) in a dose-dependent manner. There were significant differences in the proliferation inhibition rate and the AI between AE medium and high dose groups and 3-MA + AE group( P < 0. 05). Compared with those in the control group,the cell autophage and autophagic vacuoles increased,the IOD/area values of lysosomes,p53,AMPK and Beclin-1 expressions significantly increased,and the m TOR expression and LC3 I/LC3 II ratio significantly decreased in AE groups( P < 0. 05) in a dose-dependent manner. The cell autophage and autophagic vacuoles decreased,the IOD/area values of lysosomes,and p53,AMPK and Beclin-1 expressions significantly decreased,and the m TOR expression and LC3 I/LC3 II ratio significantly increased in 3-MA + AE group( P < 0. 05). Conclusion: AE can induce the apoptosis of SGC-7901 cells by increasing the autophagy level of SGC-7901 cells. The mechanism may be related to promoting the expression of p53/AMPK/m TOR signaling pathway.
作者
刘豪杰
陈文礼
陈雪蕾
Liu Haojie;Chen Wenli;Chen Xuelei(Department of Gastroenterology, Ezhou Central Hospital, Hubei Ezhou 436000, China;Department of Gastroenterology, the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine)
出处
《中国药师》
CAS
2019年第10期1829-1834,共6页
China Pharmacist
