摘要
目的:研究自噬抑制剂3-甲基腺苷(3-MA)对姜黄素诱导喉癌Hep-2细胞系自噬和凋亡的影响。方法:常规培养人喉癌Hep-2细胞,MTT法检测不同浓度姜黄素对细胞的增殖抑制率;AO吖啶橙染色法观察细胞的自噬水平,Annexin VFITC/PI双染法流式细胞仪检测各组细胞凋亡率;Western blot检测LC3、Beclin1、Bcl-2以及Bax蛋白的表达水平。结果:MTT法结果显示姜黄素呈时间-剂量依赖性抑制Hep-2细胞的增殖;姜黄素联合使用3-MA后可使凋亡率增多(P<0.05)。吖啶橙染色结果显示3-MA联合姜黄素能明显降低姜黄素引起的喉癌Hep-2细胞的自噬水平。Western blot结果显示联合使用3-MA后,蛋白Bcl-2以及Bclin-1表达降低,Bax蛋白表达水平升高,LC3Ⅱ表达减少(P<0.05)。结论:姜黄素能诱导喉癌Hep-2细胞凋亡,同时也诱导了保护性自噬的发生。自噬抑制剂3-MA抑制自噬后,能明显增强姜黄素对喉癌Hep-2细胞系的凋亡诱导效应。
Objective：The aim of this study is to study the effect of 3-methytadenine（3-MA） on the autophagy and apoptosis of human laryngeal cancer Hep 2 cells induced by curcumin. Method：The proliferation of human la- ryngeal 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, Beclinl, Bcl-2 and Bax protein were detected by Western blot. Result.. MTT assay showed that curcumin in- hibited the proliferation of Hep2 cells in a dose/time dependent manner. The apoptosis rate of curcumin combined with 3-MA increased（P〈0.05）. Acridine orange staining showed that 3-MA combined with curcumin could sig- nificantly 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（P〈0.05）. Conclusion： 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.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
北大核心
2017年第10期785-788,共4页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
关键词
姜黄素
自噬
凋亡
喉癌
curcumin
autophagy
apoptosis
human laryngeal cancer
