AMPK/mTOR信号通路介导的自噬在血根碱抑制鼻咽癌细胞增殖中的作用研究  

Role of autophagy in sanguinarine inhibiting nasopharyngeal carcinoma cells proliferation via AMPK/mTOR signaling pathway

作　　者：苏俐丹 何迎春(指导)[1] 黄立中[2] 刘洁[1] SU Lidan;HE Yingchun;HUANG Lizhong;LIU Jie(Hunan Provincial Key Laboratory for Prevention and Treat-ment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医药防治眼耳鼻咽喉疾病湖南省重点实验室,长沙410208 [2]湖南中医药大学中西医结合学院,长沙140208

出　　处：《中国免疫学杂志》2022年第23期2870-2875,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(81874408);湖南省中医药管理局项目(2021014);湖南中医药大学中西医结合一流学科开放基金项目(2018ZXYJH34)。

摘　　要：目的:研究血根碱(SAN)对人鼻咽癌5-8F细胞自噬和增殖的影响,并探讨5-8F细胞自噬与增殖的关系及作用机制。方法:MTT和实时无标记细胞分析技术(RTCA)检测细胞增殖;单丹磺酰戊二胺(MDC)染色和透射电镜观察自噬情况;Western blot检测蛋白表达。结果:与Control组相比,SAN可显著抑制鼻咽癌细胞增殖,降低PCNA表达,诱导自噬小体形成,降低P62表达,提高Beclin-1表达、LC3Ⅱ/Ⅰ;而抑制自噬后(3-MA预处理),SAN诱导自噬和抑制细胞增殖作用显著减弱,同时,SAN对Beclin-1、LC3Ⅱ/Ⅰ、p62、PCNA的影响改变;SAN可激活AMPK/mTOR信号通路关键蛋白AMPK、p-AMPK并抑制mTOR表达;而抑制AMPK/mTOR信号通路后(抑制剂Compound C预处理),SAN诱导自噬和抑制细胞增殖作用显著减弱。结论:SAN能够抑制人鼻咽癌5-8F细胞增殖并诱导自噬,可能通过诱导细胞自噬抑制细胞增殖,其机制可能与激活AMPK/mTOR信号通路有关。Objective:To investigate effect of sanguinarine(SAN)on proliferation and autophagy in nasopharyngeal carcinoma 5-8F cells,and to explore relationship between autophagy and proliferation of 5-8F cells and its mechanism.Methods:MTT and real time cellular analysis(RTCA)were used to detect cell proliferation;Monodansylcadaverine(MDC)staining and transmission electron microscope were used to observe autophagy;Western blot was used to detect protein expression.Results:Compared with Control group,SAN could significantly inhibit proliferation of nasopharyngeal carcinoma cells and reduce expression of PCNA,induce formation of autophagosomes,reduce P62 expression and increase Beclin-1 expression and LC3Ⅱ/Ⅰ.After inhibiting autophagy(3-MA pre-treatment),SAN’s effects of inducing autophagy and inhibiting cell proliferation were significantly reduced.At the same time,SAN had a different effect on Beclin-1,LC3Ⅱ/Ⅰ,p62 and PCNA.SAN conld activate AMPK/mTOR signaling pathway key proteins AMPK,p-AMPK and inhibit expression of mTOR;and after inhibiting AMPK/mTOR signaling pathway(pretreatment with inhibitor Compound C),SAN’s effect of inducing autophagy and inhibiting cell proliferation was significantly reduced.Conclusion:SAN can inhibit proliferation of human nasopharyngeal carcinoma 5-8F cells and induce autophagy,may inhibit cell proliferation by inducing autophagy,whose mechanism may be related to activation of AMPK/mTOR signaling pathway.

关 键 词：血根碱 鼻咽癌 自噬 增殖 AMPK/mTOR信号通路 

分 类 号：R739.62[医药卫生—肿瘤]