天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (2): 143-145.doi: 10.11958/20160975

• 细胞与分子生物学 • 上一篇    下一篇

白黎芦醇对鼻咽癌 CNE-2 细胞自噬的影响及机制

王敏， 赵红， 郑家法， 谢远杰， 姚平波   

  1. 南华大学附属南华医院耳鼻喉科 （邮编 441002）
  • 收稿日期:2016-09-07 修回日期:2016-11-08 出版日期:2017-02-15 发布日期:2017-02-14
  • 作者简介:王敏 （1983）， 男， 硕士， 主治医师， 主要从事鼻咽部肿瘤研究
  • 基金资助:
    湖南省医药卫生基金资助项目 （B2013-037）

The effect and mechanism of resveratrol on autophagy regulation in nasopharyngeal carcinoma CNE-2 cells

WANG Min, ZHAO Hong, ZHENG Jia-fa, XIE Yuan-jie, YAO Ping-bo   

  1. Department of Otorhinolaryngology, South China Affiliated Hospital of South China University, Hengyang 441002, China
  • Received:2016-09-07 Revised:2016-11-08 Published:2017-02-15 Online:2017-02-14

PDF (PC)

3562

摘要： 目的 观察白藜芦醇（RES）对鼻咽癌 CNE-2 细胞自噬活性及自噬流的影响， 并探讨其作用机制。方法取人低分化鼻咽癌细胞系 CNE-2， 分为对照组、 RES 干预组， 对照组置于 37 ℃培养箱中培养， 不作处理； RES 干预组加入 40 μmol/L RES， 置 37 ℃培养箱中培养。采用 Western blot 法检测 2 组自噬微管相关蛋白轻链 3B （LC3B）、 p62、 Beclin-1 及 mTOR 通路蛋白 p-mTOR、 p-S6 表达水平， 腺病毒 GFP-mRFP-LC3 荧光瞬时转染技术通过不同颜色斑点检测自噬流表达。结果 （1） RES 干预组 LC3B 蛋白表达水平明显高于对照组， p62 蛋白表达水平低于对照组 （均 P＜0.05）。2 组 Beclin-1 蛋白表达差异无统计学意义。（2） RES 干预组 p-mTOR 和 p-S6 蛋白表达水平明显低于对照组 （均 P＜0.05）。（3） RES 干预组的 mRFP 荧光点数 （红点） 多于对照组， GFP 荧光点数 （黄点） 则少于对照组 （均 P＜ 0.05）。结论 RES 能够促进鼻咽癌 CNE-2 细胞自噬流， 其机制可能与促进 mTOR 信号通路中的相关蛋白表达有关。

关键词: 鼻咽肿瘤, 自噬, 白藜芦醇, CNE-2

Abstract: Objective To investigate the effect of resveratrol on autophagic flux of nasopharyngeal carcinoma CNE-2 cells, and to explore the underlying mechanism. Methods Nasopharyngeal carcinoma CNE- 2 cells were divided into control group and resveratrol group. Cells in control group were normally cultured at 37 ℃ and received no further treatment. Resveratrol group was added 40 μmol/L resveratrol 2 h before cells were culture at 37 ℃ . Western blot analysis was performed to detect protein expressions of LC3B, p62, Beclin-1, phospho-mTOR (p-mTOR) and phospho-S6 (p-S6). The autophagic flux was detected under the confocal laser scanning microscopy through different color spots, after cells were transfected with adenovirus encoding GFP- mRFP- LC3. Results (1) The protein expression of LC3B was significantly increased and the protein expression of p62 was significantly decreased in resveratrol group compared with those of control group (P＜0.05). There was no significant difference in Beclin- 1 expression between two groups. (2) Compared to control group, expressions of p- mTOR and p- S6 were significantly decreased in resveratrol group (P＜0.05). (3) Compared to control group, the red mRFP puncta were significantly increased, and the yellow GFP puncta were significantly decreased in resveratrol group (P＜0.05). Conclusion Resveratrol promotes the autophagic flux of nasopharyngeal carcinoma CNE- 2 cells, and the effects are possibly dependent on the activation of mTOR pathway-related proteins.