AMPK在维生素E琥珀酸酯诱导人胃癌细胞自噬中的作用研究

doi:10.3969/j.issn.1004-616x.2016.05.002

癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (5): 342-347.doi: 10.3969/j.issn.1004-616x.2016.05.002

AMPK在维生素E琥珀酸酯诱导人胃癌细胞自噬中的作用研究

余洋, 侯丽颖, 宋华翠, 吴坤

哈尔滨医科大学公共卫生学院营养与食品卫生学教研室, 黑龙江哈尔滨 150081

收稿日期:2016-01-11 修回日期:2016-06-16 出版日期:2016-09-30 发布日期:2016-09-30
通讯作者: 吴坤,E-mail:wukun_15000@126.com E-mail:wukun_15000@126.com
作者简介:余洋,E-mail:909865119@qq.com
基金资助:
国家自然科学基金资助项目（81172651）

Involvement of AMPK in vitamin E succinate-induced autophagy in human gastric cancer cells

YU Yang, HOU Liying, SONG Huacui, WU Kun

Department of Nutrition and Food Hygiene, School of Public Health, Harbin Medical University, Harbin 150081, Heilongjiang, China

Received:2016-01-11 Revised:2016-06-16 Online:2016-09-30 Published:2016-09-30

摘要/Abstract

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、20 μg/mL）VES处理SGC-7901细胞24 h，以及20 μg/mL 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，20 μg/mLVES处理SGC-7901细胞24 h，Western blot法检测LC3、Beclin-1蛋白以及AMPK、mTOR、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 mRNA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 mRNA表达下降（P<0.05）；mTOR、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mTOR及其下游分子活性，继而诱导自噬的发生。

关键词: 维生素E琥珀酸酯, 腺苷酸活化蛋白激酶, 自噬, 胃癌细胞

Abstract:

OBJECTIVE: To study the role of AMPK in autophagy which was induced by vitamin E succinate (VES) in human gastric cancer SGC-7901 cells. METHODS: SGC-7901 cells were treated with different doses (5,10,15,20 μg/mL) of VES or with 20 μg/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),Beclin-1 and the phosphorylation level of AMPK. RNAi was used to silence AMPK expression in SGC-7901 cells,then these cells were treated with 20 μg/mL VES for 24 h. Western blot was used to evaluate the protein levels of LC3,Beclin-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 Beclin-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,Beclin-1 and LC3 mRNA were decreased,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.

Key words: vitamin E succinate, AMPK, autophagy, gastric cancer cell line

中图分类号:

R730.2

余洋, 侯丽颖, 宋华翠, 吴坤. AMPK在维生素E琥珀酸酯诱导人胃癌细胞自噬中的作用研究[J]. 癌变·畸变·突变, 2016, 28(5): 342-347.

YU Yang, HOU Liying, SONG Huacui, WU Kun. Involvement of AMPK in vitamin E succinate-induced autophagy in human gastric cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(5): 342-347.

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AMPK在维生素E琥珀酸酯诱导人胃癌细胞自噬中的作用研究

Involvement of AMPK in vitamin E succinate-induced autophagy in human gastric cancer cells

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