芝麻酚通过AMPK/SIRT1/NF-κB信号通路调控食管鳞状细胞癌Eca109细胞的自噬和凋亡  

Sesamol regulates autophagy and apoptosis of esophageal squamous cell carcinoma Eca109 cells through AMPK/SIRT1/NF-κB signal pathway

作　　者：刘山 王华兵 刘冲 张倬 LIU Shan;WANG Huabing;LIU Chong;ZHANG Zhuo(Department of Thoracic Surgery,Wuhan Third Hospital,Wuhan 430060,Hubei,China;Intensive Care Unit,Wuhan Third Hospital,Wuhan 430060,Hubei,China)

机构地区：[1]武汉市第三医院胸外科,湖北武汉430060 [2]武汉市第三医院重症监护室,湖北武汉430060

出　　处：《中国肿瘤生物治疗杂志》2023年第2期123-128,共6页Chinese Journal of Cancer Biotherapy

基　　金：湖北省科技厅科研计划项目(No.2019CFB789)。

摘　　要：目的:探讨芝麻酚(SEM)通过腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子1(SIRT1)/核因子κB(NF-κB)通路影响食管鳞状细胞癌(ESCC)Eca109细胞自噬和凋亡的机制。方法:用不同浓度的SEM(0、1.5625、3.125、6.25、12.5、25、50、100、200、400μmol/L)分别处理Eca109细胞、人食管上皮细胞HEEpiC 48 h,CCK-8法检测细胞增殖率,筛选适宜的SEM浓度用于后续实验。将Eca109细胞分为对照组(CK组,0μmol/L)、低剂量SEM组(SEM-L组,25μmol/L)、中剂量SEM组(SEM-M组,50μmol/L)、高剂量SEM组(SEM-H组,100μmol/L)、高剂量SEM+Compound C(AMPK抑制剂)组(SEM-H+Compound C组,100μmol/L+10μmol/L),所有各组Eca109细胞在对应的药物浓度下处理48 h后,CCK-8法检测Eca109细胞增殖,流式细胞术检测细胞凋亡,透射电镜观察Eca109细胞内自噬小体,WB法检测Eca109细胞中微管相关蛋白1轻链3(LC3)-Ⅱ/LC3-Ⅰ、Beclin-1、B淋巴细胞瘤2(Bcl2)、Bcl2相关X蛋白(BAX)、p-AMPK、SIRT1、p-NF-κB p65的表达。结果:通过预实验选择SEM实验浓度为25、50、100μmol/L用于正式研究。在SEM处理下,与CK组比较,SEM-L组、SEM-M组、SEM-H组Eca109细胞的增殖水平(24、48 h)和Bcl2、p-NF-κB p65蛋白表达均显著降低,细胞凋亡率和自噬小体数量、LC3-Ⅱ/LC3-Ⅰ、Beclin-1、BAX、p-AMPK、SIRT1蛋白表达显著升高,且呈剂量依赖性(均P<0.05);与SEM-H组比较,SEM-H+Compound C组Eca109细胞增殖水平(24、48 h)和Bcl2、p-NF-κB p65蛋白表达均显著升高,细胞凋亡率和自噬小体数量、LC3-Ⅱ/LC3-Ⅰ、Beclin-1、BAX、p-AMPK、SIRT1蛋白表达均显著降低(均P<0.05)。结论:SEM可能通过激活AMPK/SIRT1信号通路而抑制NF-κB活性来促进Eca109细胞自噬与凋亡。Objective:To explore the effect of sesamol(SEM)on autophagy and apoptosis of esophageal squamous cell carcinoma(ESCC)Eca109 cells through adenylate activated protein kinase(AMPK)/silencing information regulator 1(SIRT1)/nuclear factorκB(NF-κB)pathway.Methods:Eca109 cells of ESCC and HEEpiC of human esophageal epithelial cells were treated with different concentrations of SEM(0,1.5625,3.125,6.25,12.5,25,50,100,200,and 400μmol/L)for 48 hours.Cell viability was detected by CCK-8 method and appropriate SEM concentrations were screened for subsequent experiments.Eca109 cells were grouped into control group(CK group,0μmol/L),low-dose SEM group(SEM-L group,25μmol/L),medium-dose SEM group(SEM-M group,50μmol/L),high-dose SEM group(SEM-M group,50μmol/L),high-dose SEM group(SEM-H group,100μmol/L)and high-dose SEM+compound C(AMPK inhibitor)group(SEM-H+Compound C group,100μmol/L+10μmol/L).All Eca109 cells were treated at the corresponding drug concentration for 48 hours.CCK-8 assay was applied to detect the proliferation of Eca109 cells;flow cytometry was applied to detect apoptosis;transmission electron microscopy was applied to observe autophagosomes in Eca109 cells and WB assay was applied to detect the protein expressions of microtubule-associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰ,Beclin-1,B-lymphoma-2(Bcl2),Bcl2-associated X protein(BAX),p-AMPK,SIRT1,p-NF-κB p65 in Eca109 cells 2.Results:The SEM concentrations of 25,50 and100μmol/L selected through pre-experiment are used for formal research.Under SEM processing,compared with the CK group,the proliferation levels(24,48 h)of Eca109 cells,protein expressions of Bcl2 and p-NF-κB p65 in SEM-L group,SEM-M group and SEM-H group decreased significantly while the apoptosis rate,the number of autophagosomes,the protein expressions of LC3Ⅱ-/LC3-Ⅰ,Beclin-1,BAX,p-AMPK and SIRT1 increased significantly,in a dose-dependent manner(all P<0.05);compared with the SEM-H group,the proliferation level(24,48 h)of Eca109 cells,protein expressions of Bcl2 and p-NF-κB p6

关 键 词：食管鳞状细胞癌 ECA109细胞 芝麻酚 腺苷酸活化蛋白激酶通路 沉默信息调节因子1通路 核因子κB通路 细胞自噬 凋亡 

分 类 号：R735.1[医药卫生—肿瘤] R730.2[医药卫生—临床医学]