【摘要】：目的:研究自噬抑制劑3-甲基腺苷(3-MA)對姜黃素誘導喉癌Hep-2細胞系自噬和凋亡的影響。方法:常規(guī)培養(yǎng)人喉癌Hep-2細胞,MTT法檢測不同濃度姜黃素對細胞的增殖抑制率;AO吖啶橙染色法觀察細胞的自噬水平,Annexin VFITC/PI雙染法流式細胞儀檢測各組細胞凋亡率;Western blot檢測LC3、Beclin1、Bcl-2以及Bax蛋白的表達水平。結果:MTT法結果顯示姜黃素呈時間-劑量依賴性抑制Hep-2細胞的增殖;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細胞的自噬水平。Western blot結果顯示聯(lián)合使用3-MA后,蛋白Bcl-2以及Bclin-1表達降低,Bax蛋白表達水平升高,LC3Ⅱ表達減少(P0.05)。結論:姜黃素能誘導喉癌Hep-2細胞凋亡,同時也誘導了保護性自噬的發(fā)生。自噬抑制劑3-MA抑制自噬后,能明顯增強姜黃素對喉癌Hep-2細胞系的凋亡誘導效應。
[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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