Wan B L, Yan T T, Zhang Y J, Li X T
Department of Otolaryngology, Henan Provincial People's Hospital, Zhengzhou, 450000, China.
Department of Pharmacy, Zhengzhou University.
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.
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.
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。
