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中国癌症防治杂志

中国癌症防治杂志 ›› 2014, Vol. 6 ›› Issue (1): 1-6.doi: 10.3969/j.issn.1674-5671.2014.01.01

• 基础研究 •    下一篇

大黄素乙酰化物致鼻咽癌CNE-1细胞线粒体自噬活性的研究

  

  1. 广西医科大学药学院；广西肿瘤防治研究所实验研究部
  • 出版日期:2014-03-25 发布日期:2014-04-10
  • 通讯作者: 候华新 E-mail:houhuaxin@163.com
  • 基金资助:

    国家自然科学基金资助项目（81060270）；广西医学科学实验中心开放基金资助项目（KFJJ2011-25）

Mitophagy of nasopharyngeal carcinoma CNE-1 cells induced by acetylated emodin derivative

  • Online:2014-03-25 Published:2014-04-10

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被引次数

3

摘要： 目的 研究大黄素乙酰化物作用鼻咽癌CNE-1细胞后，其线粒体损伤及其与线粒体自噬的关系。方法 以5 mg·L-1及10 mg·L-1大黄素乙酰化物作用鼻咽癌CNE-1细胞，通过透射电镜观察大黄素乙酰化物导致鼻咽癌CNE-1 细胞自噬体形成的超微结构；激光共聚焦显微镜测定线粒体膜电位、细胞内钙离子、细胞内活性氧的浓度变化及线粒体自噬的活性。结果 与空白对照组比较，两个实验组细胞都出现线粒体损伤以及线粒体自噬体。大黄素乙酰化物作用后的细胞线粒体膜电位降低（P<0.05）；细胞内游离的钙离子浓度升高（P<0.05）；细胞内活性氧增加（P<0.05），且呈剂量依赖性。经线粒体与溶酶体示踪分析，可见大黄素乙酰化物作用的实验组均出现酸性溶酶体增殖，其中包含线粒体探针标记的线粒体碎片等典型的自噬特征，且增加的趋势呈剂量效应关系。结论 大黄素乙酰化物可致鼻咽癌CNE-1细胞线粒体损伤，并与线粒体自噬密切相关。

关键词: 鼻咽肿瘤, 大黄素乙酰化物, 线粒体自噬, 透射电镜, 激光共聚焦显微镜

Abstract: Objective To explore the relationship between mitochondrial damage and mitophagy in nasopharyngeal carcinoma CNE-1 cells treated with acetylated emodin derivative. Methods CNE-1 cells were divided into a control group and groups treated with 5 or 10 mg·L-1 of acetylated emodin derivative.After treatment， autophagosome ultrastructure was analyzed by transmission electron microscopy，while mitochondrial membrane potential and levels of intracellular free calcium ion and reactive oxygen species were measured by confocal microscopy. Results Cells treated with acetylated emodin derivative showed mitochondrial damage and the presence of mitophagosomes，which were not observed in control cells.The emodin derivative also led to a significant decrease in mitochondrial membrane potential and increases in levels of intracellular free calcium ion and reactive oxygen species（P<0.05）；these effects were dose-dependent. Analyzing mitochondrial and lysosome markers of treated cells revealed typical mitophagic features， including acidic lysosomal proliferation and lysosomes containing mitochondrial remnants labeled with Mito Tracker Green. The abundance of these features was also dose-dependent. Conclusion The acetylated emodin derivative can cause mitochondrial damage in nasopharyngeal carcinoma CNE-1 cells，perhaps through a mechanism closely related to mitophagy.

Key words: Nasopharyngeal neoplasm, Acetylated emodin derivative, Mitophagy, Transmission electron microscopy, Confocal laser scanning microscopy