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自噬在姜黄素诱导人喉癌Hep2细胞增殖中的作用

[The role of autophagy in the curcumin induced proliferation in human laryngeal cancer Hep2 cell].

作者信息

Wan B L, Yan T T, Zhang Y J, Li X T

机构信息

Department of Otolaryngology, Henan Provincial People's Hospital, Zhengzhou, 450000, China.

Department of Pharmacy, Zhengzhou University.

出版信息

Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2017 May 20;31(10):785-788. doi: 10.13201/j.issn.1001-1781.2017.10.012.

Abstract

The aim of this study is to study the effect of 3-methyladenine (3-MA) on the autophagy and apoptosis of human laryngeal cancer Hep 2 cells induced by curcumin. The proliferation of human laryngeal cancer Hep2 cells was observed by MTT assay. The autophagy level was detected by AO acridine orange staining. Annexin VFITC/PI double staining was used to detect the apoptosis of Hep2 cells. The expression of LC3, Beclin1, Bcl-2 and Bax protein were detected by Western blot. MTT assay showed that curcumin inhibited the proliferation of Hep2 cells in a dose/time dependent manner. The apoptosis rate of curcumin combined with 3-MA increased (<0.05). Acridine orange staining showed that 3-MA combined with curcumin could significantly reduce the autophagy level of laryngeal carcinoma Hep2 cells. The expression of Bcl-2, Bclin-1 and LC3 Ⅱ was decreased, while the expression of Bax protein was increased (<0.05). Curcumin can induce apoptosis of Hep 2 cells and induce the development of protective autophagy. The inhibitory effect of curcumin on the apoptosis of laryngeal carcinoma Hep2 cell line was significantly enhanced by 3-MA.

摘要

本研究旨在探讨3-甲基腺嘌呤(3-MA)对姜黄素诱导的人喉癌Hep 2细胞自噬和凋亡的影响。采用MTT法观察人喉癌Hep2细胞的增殖情况。通过吖啶橙染色检测自噬水平。采用Annexin V-FITC/PI双染法检测Hep2细胞的凋亡情况。通过蛋白质免疫印迹法检测LC3、Beclin1、Bcl-2和Bax蛋白的表达。MTT法显示,姜黄素以剂量/时间依赖性方式抑制Hep2细胞的增殖。姜黄素联合3-MA的凋亡率增加(<0.05)。吖啶橙染色显示,3-MA联合姜黄素可显著降低喉癌Hep2细胞的自噬水平。Bcl-2、Bclin-1和LC3Ⅱ的表达降低,而Bax蛋白的表达增加(<0.05)。姜黄素可诱导Hep 2细胞凋亡并诱导保护性自噬的发生。3-MA可显著增强姜黄素对喉癌Hep2细胞系凋亡的抑制作用。

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