EBSS饥饿人鼻咽癌CNE-2细胞株诱导自噬的机制研究  

Research of EBSS starvation induced autophagy and its mechanism in CNE-2 cells

作　　者：刘芮吟 吴梦鸽 曾汶 张睿[1] 李诗琴[1] 费继敏[1] Liu Ruiyin;Wu Mengge;Zeng Wen;Zhang Rui;Li Shiqin;Fei Jimin(Department of Head and Neck,Third Affiliated Hospital of Kunming Medical,Yunnan Kunming 650118,China)

机构地区：[1]昆明医科大学第三附属医院云南省肿瘤医院头颈外科,云南昆明650118

出　　处：《现代肿瘤医学》2019年第1期30-35,共6页Journal of Modern Oncology

基　　金：云南省卫生高层次人才基金资助项目(编号:D-201659)

摘　　要：目的:观察饥饿状态下永生化正常鼻咽上皮NP69细胞、鼻咽癌CNE-2细胞的自噬变化及自噬与凋亡的相互作用,为Baf-A1在鼻咽癌中的应用前景提供证据。方法:EBSS溶液替代DMEM溶液在NP69和CNE-2细胞中制造体外饥饿环境,Baf-A1抑制细胞自噬。应用Western blot及透射电镜检测细胞自噬,MTT检测细胞存活率,qRT-PCR检测mRNA表达。结果:与NP69细胞相比,EBSS制造的饥饿环境显著促进CNE-2细胞发生自噬,处理24时后出现大量细胞凋亡,100 nmol/L Baf-A1抑制细胞自噬可延缓饥饿诱导的细胞凋亡。qRT-PCR检测结果:EBSS作用下,LKB1、AMPK、ULK1、Beclin1、ATG5 mRNA水平均显著上升(P <0. 05)。结论:EBSS通过LKB1/AMPK/m TOR通路显著诱导CNE-2细胞自噬的发生。Objective:To investigate starvation induced autophagy and mechanism in normal nasopharyngeal cell line NP69 and nasopharyngeal carcinoma CNE-2 cell,to provide evidence for the application of Baf-A1 in nasopharyngeal carcinoma.Methods:NP69 and CNE-2 cell were starvation induced by EBSS medium instead of DMEM medium,Baf-A1 inhibit autophagy of CNE-2 cell.Western blot were used to detect the special markers of autophagic microtubule-associated protein 1 light chain 3(LC3),and transmission electron microscope(TEM)was used to observe the autophagic body.In condition,cell proliferation and apoptosis induced by nutrient depletion was measured by MTT assay.qRT-PCR analysis of the mRNA expression of autophagy related genes.Results:Compared with NP69,CNE-2 cell autophagy activity was significantly promoted in stavation condition induced by EBSS solution.The cell apoptosis was delayed by 100 nmol/L Baf-A1 which inhibit autophagy of CNE-2 cell.Meanwhile,the results of qRT-PCR demonstrated EBSS significantly promoted mRNA levels of LKB1,AMPK,ULK1,Beclin1,ATG5 in CNE-2 cells(P<0.05).Conclusion:The starvation condition may induce autophagy of CNE-2 cells by LKB1/AMPK/mTOR pathway.

关 键 词：鼻咽癌 自噬 饥饿 LC3 LKB1 BECLIN1 

分 类 号：R739.6[医药卫生—肿瘤]