中医学报2024，Vol.39Issue(6)：1280-1288,9.DOI:10.16368/j.issn.1674-8999.2024.06.215
基于EGFR/PI3K/AKT/mTOR通路探讨表没食子儿茶素-3-没食子酸酯调控非小细胞肺癌细胞周期和凋亡的机制
Mechanism of Epigallocatechin-3-gallate Regulating Cell Cycle and Apoptosis of Non-small Cell Lung Cancer(NSCLC)Based on EGFR/PI3K/AKT/mTOR Pathway
摘要
Abstract
Objective:Based on epidermal growth factor receptor(EGFR)/phosphoinositide 3-kinase(PI3K)/protein kinase B,Dis-cussion on the AKT/mammalian target of rapamycin(mTOR)Pathway of epigallocatechin-3-gallate,EGCG regulates cell cycle and apoptosis in non-small cell lung cancer(NSCLC)A549.Methods:A549 cells were randomly divided into blank control group,model group,EGCG group,gefitinib group and EGCG+gefitinib group.The proliferation of A549 cells was detected by MTT assay,and the cell cycle and apoptosis were detected by flow cytometry.Molecular docking and cellular thermal shift assay(CETSA)were used to de-tect the targeted binding of EGCG to EGFR.cyclin-dependent kinase 4(CDK4),B-cell lymphoma-2(Bcl-2)were detected by Western blot.Expression levels of Bcl-2,Bcl2-associated X protein(Bax),p-EGFR/EGFR,p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR.Results:Compared with the model group,EGCG and gefitinib both showed significant inbibitory effects on A594 cells.Compared with the blank control group,the cell percentage in the model group was decreased in G1/G0 phase and increased in G2/M phase(P<0.01).Compared with model group,the cell percentage of EGCG group,gefitinib group and EGCG+gefitinib group was increased in G1/G0 stage(P<0.01),while the cell percentage of gefitinib group and EGCG+gefitinib group was decreased in G2/M stage(P<0.01).Compared with blank control group,apoptosis rate of model group was decreased(P<0.01).Compared with model group,the apoptosis rate of EGCG,gefitinib and EGCG+gefitinib groups was increased(P<0.05).Compared with blank con-trol group,CDK4,Bcl-2 and Bcl-2/Bax protein expression levels in model group were increased(P<0.01),while Bax protein ex-pression levels were decreased(P<0.01).Compared with model group,the expression levels of CDK4,Bcl-2 and Bcl-2/Bax in EGCG group,gefitinib group and EGCG+gefitinib group were decreased(P<0.01),while the expression levels of Bax were increased(P<0.05).Molecular docking results showed that the binding energy of EGCG and gefitinib with EGFR was less than-5.0 kcal·mol-1.The CETSA results showed that compared with RPMI 1640 group,the CETSA curve of EGFR protein in EGCG group shifted to the right.Compared with blank control group,the protein expression levels of p-EGFR/EGFR,p-PI3K/PI3K,p-Akt/AKT and p-mTOR/mTOR in model group were increased(P<0.05).Compared with model group,the protein expression levels of p-EGFR/EG-FR,p-PI3K/PI3K,p-Akt/AKT and p-mtor/mTOR in EGCG group,gefitinib group and EGCG+gefitinib group were decreased(P<0.01).Conclusion:EGCG may regulate A549 cell cycle and apoptosis through targeted regulation of EGFR/PI3K/AKT/mTOR signaling pathway,thus playing an anticancer role.关键词
表没食子儿茶素-3-没食子酸酯/非小细胞肺癌/A549细胞/EGFR/PI3K/AKT/mTOR通路/细胞周期/细胞凋亡Key words
epigallocatechin-3-gallate/non-small cell lung cancer/A549 cell/EGFR/PI3K/AKT/mTOR pathway/cell cycle/cell apoptosis分类
医药卫生引用本文复制引用
任闪闪,刘燕,尚艺婉,吴耀松,刘俊,陈玉龙..基于EGFR/PI3K/AKT/mTOR通路探讨表没食子儿茶素-3-没食子酸酯调控非小细胞肺癌细胞周期和凋亡的机制[J].中医学报,2024,39(6):1280-1288,9.基金项目
国家自然科学基金项目(81960039) （81960039）
河南中医药大学科研苗圃工程项目(MP2020-18) （MP2020-18）
