自噬在姜黄素诱导喉癌Hep-2细胞死亡中的作用
发布时间：2018-10-23 14:32
【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(P0.05)。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(P0.05)。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。
[Abstract]:Aim: to study the effects of 3 methyl adenosine (3-MA), an autophagy inhibitor, on autophagy and apoptosis of Hep-2 cell line induced by curcumin. Methods: Hep-2 cells of human laryngeal carcinoma were cultured routinely, the proliferation inhibition rate of different concentrations of curcumin was detected by MTT assay, the autophagy level was observed by AO acridine orange staining, and the apoptosis rate; Western blot was detected by Annexin VFITC/PI double staining flow cytometry. The expression level of LC3,Beclin1,Bcl-2 and Bax protein was detected. Results: MTT assay showed that curcumin inhibited the proliferation of Hep-2 cells in a dose-dependent manner, and curcumin combined with 3-MA increased the apoptosis rate (P0.05). The results of acridine orange staining showed that 3-MA combined with curcumin could significantly reduce the autophagy level of Hep-2 cells induced by curcumin. The results showed that after combined use of 3-MA, the expression of protein Bcl-2 and Bclin-1 decreased, the expression of Bax protein increased, and the expression of LC3 鈪，
本文编号：2289528
[Abstract]:Aim: to study the effects of 3 methyl adenosine (3-MA), an autophagy inhibitor, on autophagy and apoptosis of Hep-2 cell line induced by curcumin. Methods: Hep-2 cells of human laryngeal carcinoma were cultured routinely, the proliferation inhibition rate of different concentrations of curcumin was detected by MTT assay, the autophagy level was observed by AO acridine orange staining, and the apoptosis rate; Western blot was detected by Annexin VFITC/PI double staining flow cytometry. The expression level of LC3,Beclin1,Bcl-2 and Bax protein was detected. Results: MTT assay showed that curcumin inhibited the proliferation of Hep-2 cells in a dose-dependent manner, and curcumin combined with 3-MA increased the apoptosis rate (P0.05). The results of acridine orange staining showed that 3-MA combined with curcumin could significantly reduce the autophagy level of Hep-2 cells induced by curcumin. The results showed that after combined use of 3-MA, the expression of protein Bcl-2 and Bclin-1 decreased, the expression of Bax protein increased, and the expression of LC3 鈪，
本文编号：2289528
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