漆黄素靶向EGFR诱导非小细胞肺癌细胞周期阻滞和凋亡  

Fisetin induces cell cycle arrest and apoptosis of non-small cell lung cancer by targeting EGFR

作　　者：任闪闪[1] 刘燕[1] 尚艺婉 吴耀松[1] 刘俊 陈玉龙[1] REN Shan-Shan;LIU Yan;SHANG Yi-Wan(Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China)

机构地区：[1]河南中医药大学中医药科学院,河南郑州450046

出　　处：《中国老年学杂志》2025年第7期1644-1650,共7页Chinese Journal of Gerontology

基　　金：河南省科技厅河南省重点研发与推广专项(科技攻关)(232102310443);2022年河南省科技研发计划联合基金(优热学科培育类)项目(222301420084);河南中医药大学科研苗圃工程(MP2020-18)。

摘　　要：目的旨在探讨漆黄素对A549细胞的增殖作用及作用机制。方法取A549细胞,随机分成空白对照组、EGF诱导模型组(10 ng/ml EGF)、漆黄素组(10 ng/ml EGF和10μg/ml漆黄素)、吉非替尼组(10 ng/ml EGF和0.5μg/ml吉非替尼)、漆黄素和吉非替尼联合用药组(10 ng/ml EGF、10μg/ml漆黄素和0.5μg/ml吉非替尼)。流式细胞术测定细胞周期及凋亡情况,分子对接、细胞热转移试验(CETSA)及免疫荧光试验测定漆黄素与表皮生长因子受体(EGFR)靶向结合情况,Western印迹测定EGFR、磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)及哺乳动物雷帕霉素靶蛋白(mTOR)表达。结果与空白对照组比较,EGF诱导模型组A549凋亡率显著降低(P<0.05);与EGF诱导模型组比较,漆黄素组A549细胞凋亡率明显增多(P<0.01)。EGF诱导模型组A549细胞以阻滞于S期为主,漆黄素组细胞以阻滞于G0/G1期为主。与空白对照组比较,EGF诱导模型组EGFR/PI3K/AKT/mTOR蛋白表达量显著增加(P<0.05,P<0.01),漆黄素与EGFR的靶向联合作用显著抑制EGFR/PI3K/AKT/mTOR蛋白表达(P<0.05)。结论漆黄素可以通过靶向调控EGFR-PI3K-AKT-mTOR信号通路,促使EGF诱导的A549细胞发生G0/G1细胞阻滞和凋亡,从而达到抗癌的目的。Objective To investigate the effects of fisetin on the proliferation of A549 cell and the underlying mechanism.Methods A549 cells were randomly divided into the blank control group,the EGF induced model group(10 ng/ml EGF),fisetin group(10 ng/ml EGF+10μg/ml fisetin),Gefitinib group(10 ng/ml EGF+0.5μg/ml Gefitinib),fisetin combined with Gefitinib group(10 ng/ml EGF,10μg/ml fisetin and 0.5μg/ml Gefitinib).The cell cycle and cell apoptosis were detected by flow cytometry,molecular docking,cellular thermal shift assay(CETSA)and immunofluorescence test were used to test the targeted binding of fisetin to epidermal growth factor receptor(EGFR),Western blot was used to detect the expressions of EGFR,phosphatidylinositol-3-kinase(PI3K),protein kinase B(AKT)and mammalian target of rapamycin(mTOR).Results Compared with the blank control group,the apoptotic rate of A549 cells was significantly reduced in the EGF induced model group(P<0.05);compared with the EGF induced model group,the apoptosis rate of A549 cells in the fisetin group was significantly increased(P<0.05).A549 cells were mainly blocked in S phase in the EGF induced model group,A549 cells were mainly blocked in G0/G1 phase in the fisetin group.Compared with the blank control group,EGFR/PI3K/AKT/mTOR protein expressions in the EGF induced model group were significantly in-creased(P<0.05,P<0.01),the combination of fisetin and EGFRtargeting significantly inhibited EGFR/PI3K/AKT/mTORprotein expressions(P<0.05).Conclusions By targeting EGFR-PI3K-AKT-mTORsignaling pathway,Fisetin could induceG0/G1 cell ar-rest and apoptosis of A549 cells induced by EGF,thus achieving the purpose of anti-cancer.

关 键 词：漆黄素 非小细胞肺癌细胞A549 凋亡 表皮生长因子受体(EGFR)/磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路 

分 类 号：R734.2[医药卫生—肿瘤]