中文摘要：

目的：研究维生素E琥珀酸酯（VES）是否影响人胃癌SGC-7901细胞中腺苷酸活化蛋白激酶（AMPK）活性,并探讨AMPK在VES诱导自噬中的作用。方法：用不同剂量（5、10、15、20μg/m L）VES处理SGC-7901细胞24 h,以及20μg/m L VES分别处理SGC-7901细胞3、6、12、24 h后,Western blot法检测SGC-7901细胞中自噬标志蛋白LC3、Beclin-1及AMPK、ACC及其磷酸化蛋白的表达。用RNAi技术沉默AMPK,20μg/m LVES处理SGC-7901细胞24 h,Western blot法检测LC3、Beclin-1蛋白以及AMPK、m TOR、p70S6K与4EBP-1及其磷酸化蛋白表达,并采用荧光定量PCR法检测LC3 m RNA表达情况。结果：VES处理SGC-7901细胞后,与对照组比较,随着VES剂量的增加、作用时间的增长,AMPK磷酸化水平上升,LC3、Beclin-1蛋白表达增加（P〈0.05）。RNAi沉默AMPK后,与单纯VES处理组比较,LC3、Beclin-1蛋白表达降低,LC3 m RNA表达下降（P〈0.05）;m TOR、p70S6K及4EBP-1磷酸化活性升高（P〈0.05）。结论：VES可诱导SGC-7901细胞中AMPK活化,AMPK活化后影响m TOR及其下游分子活性,继而诱导自噬的发生。

英文摘要：

OBJECTIVE： To study the role of AMPK in autophagy which was induced by vitamin E succinate （VES） in human gastric cancer SGC-7901 cells. ME T HODS：（5,10,15,20μg/mL） of VES or with 20 [ig/mL VES for 3 ,6 ,12 ,24 h. Western blot was used to evaluate the microtubule-associated protein 1 light chain 3 （LC3,an autophagy marker protein）,Becl in-1 and the phosphorylation level of AMPK. RNAi was used to silence AMPK expression in SGC-7901 cells,then these cells were treated with 20μg/mL VES for 24 h. Western blot was used to evaluate the protein levels of LC3,Becl in -1 ,phosphorylation level of AMPK,mTOR,p70S6K and 4EBP-1. Real time PCR was used to detect the mRNA level of LC3. RESULTS ：With increasing doses of VES,and prolonged time of VES treatment,the phosphorylation level of AMPK,LC3 and Becl in-1 were elevated compared with controls （P〈0.05）. When AMPK was silenced,in VES treatment plus AMPK silenced group,both the protein level of LC3,Be cl in-1 and LC3 mRNA were d e c re a s ed ,compared with those among the non - silenced group （P〈0.05）, and the protein activity of mTOR, p70S6K and 4EBP-1 were increased （P〈0.05）. CONCLUSION： The activity of AMPK was up-regulated in VES-treated human gastric cancer SGC-7901 cells. AMPK was involved in VES-induced autophagy via inhibiting mTOR and its downstream molecular activity.