依巴斯汀通过抑制AKT/mTOR通路诱导人黑素瘤细胞自噬  

Ebastine induces autophagy in human melanoma cells by inhibiting AKT/mTOR pathway

作　　者：张平 王静[3] 王颖超 倪莉 李明明 党宁宁 ZHANG Ping;WANG Jing;WANG Yingchao;NI Li;LI Mingming;DANG Ningning(Cheeloo College of Medicine,Shandong University,Jinan 250012,China;Jinan Central Hospital,Shandong University,Jinan 250013,China;Department of Dermatology,Binzhou Medical University Hospital,Binzhou 256600,China;Shandong Provincial Hospital,Shandong First Medical University,Jinan 250021,China)

机构地区：[1]山东大学齐鲁医学院,济南250012 [2]山东大学附属济南市中心医院,济南250013 [3]滨州医学院附属医院皮肤科,滨州256600 [4]山东第一医科大学附属省立医院,济南250021

出　　处：《中国麻风皮肤病杂志》2022年第10期673-677,共5页China Journal of Leprosy and Skin Diseases

基　　金：山东省自然科学基金面上项目(编号:ZR2017MH074)。

摘　　要：目的:研究依巴斯汀对人黑素瘤细胞自噬的影响及机制。方法:体外培养人黑素瘤细胞A375和M14,采用CCK-8增殖实验检测细胞活力,并计算IC50;利用mCherry-EGFP-LC3B双荧光指示系统检测自噬流;采用Western blot验证自噬相关蛋白LC3,Beclin1及信号通路蛋白的表达。结果:依巴斯汀可显著抑制人黑素瘤细胞的活力;依巴斯汀明显诱导人黑素瘤细胞中自噬小体和自噬溶酶体的产生;依巴斯汀显著上调人黑素瘤细胞中LC3-Ⅱ/Ⅰ的比值以及Beclin1的表达,同时抑制AKT/mTOR信号通路的活化,降低p-AKT/AKT和p-mTOR/mTOR。结论:依巴斯汀通过抑制AKT/mTOR通路诱导人黑素瘤细胞自噬的发生。Objective:To study the effect and mechanism of ebastine on autophagy of human melanoma cells.Methods:The human melanoma cells A375 and M14 were cultured in vitro and cell viability was detected by a cell counting kit(CCK-8)and IC50 was calculated.The autophagy flow was detected by mCherry-EGFP-LC3B fluorescence plasmid.Western blot was used to detect the expression level of autophagy-related proteins LC3,Beclin1 and signaling pathway proteins.Results:After the action of ebastine,the cell viabilities of human melanoma cells decreased,the production of autophagosomes and autophagolysosomes in human melanoma cells increased,the ratio of LC3-Ⅱ/Ⅰand the expression of Beclin1 increased,the activation of AKT/mTOR signaling pathway decreased,and p-AKT/AKT and p-mTOR/mTOR in human melanoma cells decreased.Conclusion:Ebastine induces significant autophagy in human melanoma cells by inhibiting the AKT/mTOR signaling pathway.

关 键 词：依巴斯汀 人黑素瘤细胞 自噬 AKT/mTOR信号通路 

分 类 号：R739.5[医药卫生—肿瘤]