《中国康复理论与实践》

《中国康复理论与实践》 ›› 2015, Vol. 21 ›› Issue (08): 905-912.

• 特稿 • 上一篇    下一篇

神经酰胺通过JNK/c-Jun信号通路诱导胶质瘤细胞自噬性死亡

张露勇1，张淼2，刘师卜1，李锐3   

  1. 1.中国食品药品检定研究院食品化妆品所，北京市100044；2.中国人民解放军总医院外科药房，北京市100853；3.中日友好医院神经外科，北京市100029。作者简介：张露勇(1980-)，男，汉族，山东荣成市人，硕士，助理研究员，主要从事食品、保健食品、化妆品的功能毒理评价研究工作。通讯作者：李锐(1971-)，男，汉族，云南昆明市人，主治医师，主要从事神经相关疾病的研究。E-mail: reedleer@sina.com。
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2015-08-25 发布日期:2015-08-25

Autophagic Cell Death in Glioma Cell Induced by Ceramide through JNK/c-Jun Pathway

ZHANG Lu-yong1, ZHANG Miao2, LIU Shi-bu1, LI Rui3   

  1. 1. Institute for Food and Cosmetics Control, National Institutes for Food and Drug Control, Beijing 100044, China; 2. Department of Pharmaceutical Care, Chinese PLA General Hospital, Beijing 100853, China; 3. Department of Neurosurgery, China-Japan Friendship Hospital, Beijing 100029, China
  • Received:1900-01-01 Revised:1900-01-01 Published:2015-08-25 Online:2015-08-25

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摘要： 目的探讨神经酰胺对胶质瘤细胞87-MG和U251 自噬性死亡的作用及机制。方法采用MTT和流式细胞的方法检测不同浓度神经酰胺刺激87-MG和U251 细胞后细胞存活和凋亡的改变；电镜和Western blotting 技术检测自噬和JNK/c-Jun 信号通路的改变，JNK药理性抑制剂SP600125 特异性抑制JNK通路，观察其对神经酰胺诱导自噬死亡的影响。结果神经酰胺刺激24 h后，87-MG和U251 存活呈现时间依赖性下降(P<0.05)；相应的细胞死亡数目剂量依赖性升高(P<0.05)，但凋亡性死亡比例较低；神经酰胺刺激后镜下观察到的自噬小体计数，LC3B/LC3A和Beclin-1 的表达以及JNK/c-Jun 的磷酸化程度都增加(P<0.05)。提前给予SP600125 抑制JNK信号通路的活性后，可以阻断神经酰胺诱导的细胞自噬性死亡(P<0.05)。结论神经酰胺可诱导胶质瘤细胞87-MG和U251 出现自噬性死亡，机制可能与JNK信号通路的活化有关。

关键词: 胶质瘤, 神经酰胺, 自噬性死亡, JNK信号通路

Abstract: Objective To observe the autophagy of 87-MG and U251 glioma cells induced by ceramide and explore the possible mechanism. Methods The viability and apoptosis of 87-MG and U251 cells were detected by MTT assay and flow cytometry, respectively. Autophagic- related protein expressions of LC3B /LC3A and Beclin-1 were determined by Western blotting. The activation of JNK/c-Jun signaling pathway induced by ceramide with or without the treatment of JNK specific inhibitor SP600125 was also measured. Results 24 hours after treatment of ceramide, the growth of 87-MG and U251 cells was significantly inhibited time-dependently (P<0.05); and the number of autophagic cells increased dose-dependently (P<0.05). The levels of LC3B/LC3A and Beclin-1 significantly increased after ceramide treatment (P<0.05). JNK signaling pathway was activated in the 87-MG and U251 cells and the phosphorylation of c-Jun also increased after ceramide treatment. This activation of autophagy could be reversed by the pre-treatment of SP600125. Conclusion Ceramide may induce autophagy in 87-MG and U251 glioma cells and the mechanism may be related to the activation of JNK/c-Jun signaling pathway.

Key words: glioma cell, ceramide, autophagic cell death, JNK signaling pathway