【摘要】：目的:研究自噬抑制劑3-甲基腺苷(3-MA)對姜黃素誘導(dǎo)喉癌Hep-2細(xì)胞系自噬和凋亡的影響。方法:常規(guī)培養(yǎng)人喉癌Hep-2細(xì)胞,MTT法檢測不同濃度姜黃素對細(xì)胞的增殖抑制率;AO吖啶橙染色法觀察細(xì)胞的自噬水平,Annexin VFITC/PI雙染法流式細(xì)胞儀檢測各組細(xì)胞凋亡率;Western blot檢測LC3、Beclin1、Bcl-2以及Bax蛋白的表達(dá)水平。結(jié)果:MTT法結(jié)果顯示姜黃素呈時(shí)間-劑量依賴性抑制Hep-2細(xì)胞的增殖;姜黃素聯(lián)合使用3-MA后可使凋亡率增多(P0.05)。吖啶橙染色結(jié)果顯示3-MA聯(lián)合姜黃素能明顯降低姜黃素引起的喉癌Hep-2細(xì)胞的自噬水平。Western blot結(jié)果顯示聯(lián)合使用3-MA后,蛋白Bcl-2以及Bclin-1表達(dá)降低,Bax蛋白表達(dá)水平升高,LC3Ⅱ表達(dá)減少(P0.05)。結(jié)論:姜黃素能誘導(dǎo)喉癌Hep-2細(xì)胞凋亡,同時(shí)也誘導(dǎo)了保護(hù)性自噬的發(fā)生。自噬抑制劑3-MA抑制自噬后,能明顯增強(qiáng)姜黃素對喉癌Hep-2細(xì)胞系的凋亡誘導(dǎo)效應(yīng)。
[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，

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