青藤碱通过MAPK/ERK/mTOR通路诱导自噬抑制人食管癌TE-1细胞增殖  

Study on Inhibition of Human Esophageal Carcinoma TE-1 Cell Proliferation by Sinomenine Induced Autophagy through MAPK/ERK/mTOR Pathway

作　　者：陈耀 肖文涛 叶玉海 陈伟毅 CHEN Yao;XIAO Wentao;YE Yuhai;CHEN Weiyi(Hunan University of Medicine,Huaihua Hunan 418000,China)

机构地区：[1]湖南医药学院,湖南怀化418000

出　　处：《中医药导报》2023年第6期16-21,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：2021年度湖南省大学生创新创业训练计划项目(4314);湖南省自然科学基金项目(2022JJ50290);湖南省卫生健康委科研课题(202104081483);湖南医药学院2020年科研孵化库建设项目第二批(《青藤碱通过PI3K/Akt/mTOR信号通路诱导自噬抑制胰腺癌干细胞增殖》。

摘　　要：目的:研究青藤碱对人食管癌TE-1细胞增殖、自噬的作用和机制。方法:将TE-1细胞分成对照组、青藤碱200 mg/L组、青藤碱400 mg/L组、青藤碱800 mg/L组。对照组不做处理,青藤碱200 mg/L组、青藤碱400 mg/L组、青藤碱800 mg/L组分别予以相应浓度的青藤碱干预。光学显微镜观察细胞形态改变,Hochest染色检测活细胞数,CCK8法检测细胞抑制率,流式细胞术检测细胞凋亡情况,透射电镜观察自噬小体情况,免疫荧光检测LC3表达情况,Western blotting检测LC3-Ⅰ、LC3-Ⅱ、Beclin-1、ERK1/2、p-ERK1/2、mTOR、p-mTOR蛋白相对表达量。结果:在200、400、800 mg/L青藤碱的作用下,TE-1细胞皱缩、变圆,漂浮在培养液中,活细胞数不断减少。青藤碱200 mg/L组、青藤碱400 mg/L组、青藤碱800 mg/L组活细胞数比例显著低于对照组(P<0.01),细胞抑制率、细胞凋亡率显著高于对照组(P<0.01);青藤碱200 mg/L组、青藤碱400 mg/L组、青藤碱800 mg/L组TE-1细胞内自噬小体多于对照组;青藤碱200 mg/L组、青藤碱400 mg/L组、青藤碱800 mg/L组TE-1细胞LC3表达、LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1蛋白相对表达量均显著高于对照组(P<0.01),p-ERK1/2/ERK1/2及p-mTOR/mTOR比值均显著低于对照组(P<0.01)。结论:青藤碱能抑制人食管癌TE-1细胞增殖,其机制与抑制MAPK/ERK/mTOR通路和诱导自噬有关。Objective:To study the effect and mechanism of sinomenine on proliferation and autophagy of human esophageal cancer TE-1 cells.Methods:TE-1 cells were divided into four groups,including control group,sinomenine 200,400 and 800 mg/L groups.The control group was untreated,and the sinomenine 200 mg/L group,sinomenine 400 mg/L group,and sinomenine 800 mg/L group were treated with sinomenine at corresponding concentrations.The cell morphological changes were observed under the optical microscope,hochest staining was used to detect the number of viable TE-1 cells,CCK8 assay was used to detect the cell inhibition rate,flow cytometry was used to evaluate the TE-1 cell apoptosis,transmission electron microscopy was used to observe autophagosome in TE-1 cell,immunofluorescence was used to detect the expression of LC3,and Western blotting was used to detect the relative expression levels of LC3-I,LC3-II,Beclin-1,ERK1/2,p-ERK1/2,mTOR and p-mTOR protein.Results:Under the treatment of 200,400,800 mg/L sinomenine,TE-1 cells shrunk,became round,floated in the culture medium,and the number of living cells decreased continuously.The proportion of viable cells in sinomenine 200 mg/L group,sinomenine 400 mg/L group and sinomenine 800 mg/L group were significantly lower than that in the control group(P<0.01),and the rate of cell inhibition and apoptosis were significantly higher than that in the control group(P<0.01).The number of autophagosomes in TE-1 cells of sinomenine 200 mg/L group,sinomenine 400 mg/L group and sinomenine 800 mg/L group were more than that of control group.The expression of LC3,the ratio of LC3-II/LC3-I,and the relative expression of Beclin-1 protein in TE-1 cells of sinomenine 200 mg/L,400 mg/L,and 800 mg/L groups were significantly higher than those of the control group(P<0.01),and the ratio of p-ERK1/2/ERK1/2 and p-mTOR/mTOR were significantly lower than those of the control group(P<0.01).Conclusion:Sinomenine can inhibit the proliferation of human esophageal cancer TE-1 cells,and its mechanism is related to

关 键 词：食管癌 TE-1细胞 青藤碱 细胞增殖 细胞自噬 MAPK/ERK/mTOR通路 

分 类 号：R285.5[医药卫生—中药学]