芒柄花黄素经ERK信号通路抑制食管癌细胞的免疫逃逸机制  

Formononetin inhibits immune escape of esophageal cancer cells through the ERK signaling pathway

作　　者：郑时桢 王炜辰[1] 马黄璜 翁小琴 池志珍[1] 苏彩平 ZHENG Shizhen;WANG Weichen;MA Huanghuang(Department of Pharmacy,the People’s Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fujian,Fuzhou 350004,China)

机构地区：[1]福建中医药大学附属人民医院药剂科,福州市350004 [2]福建省中医药大学国医堂

出　　处：《河北医药》2024年第20期3045-3050,共6页Hebei Medical Journal

基　　金：福建省科技计划项目(编号:2016Y0049)。

摘　　要：目的探讨芒柄花黄素(Formononetin)通过ERK信号通路对食管癌细胞免疫逃逸的影响机制。方法选取食管癌TE-1细胞系,原代培养,选取P3代单层细胞纳入研究;采用不同浓度(0、50、100、200μg/mL)Formononetin处理细胞24 h;MTT、划痕实验、Transwell实验检测TE-1细胞增殖、迁移、侵袭能力;ELISA检测细胞上清液免疫逃逸相关因子(IL-4、IL-10、TNF-α)水平;Western-blot检测免疫逃逸关键因子B7-H1及ERK信号通路关键蛋白(ERK1/2、c-Fos、c-Jun)浓度表达;食管癌TE-1细胞采用ERK信号通路抑制剂SCH772984处理,检测Control组、Formononetin组、SCH772984组、Formononetin+SCH772984组ERK信号通路关键蛋白(ERK1/2、c-Fos、c-Jun)表达,确定Formononetin对ERK信号通路的靶向调控。结果随着细胞培养时间延长,各组食管癌细胞均体现出增殖的趋势(P<0.05);与Control组相比,Formononetin的干预可明显下调食管癌细胞的增殖能力、迁移能力及侵袭能力(P<0.05);且随着Formononetin干预剂量的提升,食管癌细胞的增殖能力、迁移能力、侵袭能力均逐渐降低,呈现计量依赖性(P<0.05)。Formononetin干预可明显下调免疫共刺激因子B7-H1、免疫逃逸相关因子(IL-4、IL-10、TNF-α)水平(P<0.05);且随着Formononetin干预剂量提升,食管癌细胞B7-H1、IL-4、IL-10、TNF-α含量均逐渐降低,呈现计量依赖性(P<0.05);Formononetin干预可明显下调ERK信号通路关键蛋白(ERK1/2、c-Fos、c-Jun)浓度表达(P<0.05);且随着Formononetin干预剂量的提升,食管癌细胞内ERK1/2、c-Fos、c-Jun浓度表达逐渐降低,呈现计量依赖性(P<0.05);与Control组相比,Formononetin、ERK信号通路抑制剂SCH772984的干预可以明显下调ERK信号通路关键蛋白(ERK1/2、c-Fos、c-Jun)浓度表达(P<0.05);二者联合干预后ERK信号通路活性进一步下降,食管癌细胞内ERK1/2、c-Fos、c-Jun浓度表达亦降低(P<0.05)。结论芒柄花黄素通过靶向抑制ERK信号通路活性降低食管癌细�Objective To explore the mechanism of Formononetin in mediating immune escape in esophageal cancer cells through the extracellular signal-regulated kinase(ERK)signaling pathway.Methods The esophageal cancer TE-1 cell line was used for primary culture.The third-generation(P3)monolayer TE-1 cells were induced with Formononetin at varying concentrations(0,50,100,200μg/mL)for 24 hours.Proliferation,migration,and invasion abilities of TE-1 cells were detected through MTT assay,wound healing assay and Transwell assay,respectively.Relative contents of immune escape-related factors(interleukin-4[IL-4],IL-10,tumour necrosis factor alpha[TNF-α])in cell supernatant were measured by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the protein expressions of the key immune escape factor B7 homolog 1(B7-H1)and ERK signaling pathway associated proteins(ERK1/2,c-Fos,c-Jun).Esophageal cancer TE-1 cells were then induced with blank control,Formononetin,the ERK signaling pathway inhibitor SCH772984,and Formononetin+SCH772984.Expression levels of key ERK signaling pathway associated proteins(ERK1/2,c-Fos,c-Jun)were detected to determine the targeted regulation of the ERK signaling pathway by Formononetin.Results With the prolongation of cell culture,TE-1 cells induced with blank control,Formononetin,SCH772984,and Formononetin+SCH772984 all presented significantly increased proliferation(P<0.05).Compared with those induced with blank control,Formononetin treatment significantly downregulated the proliferation,migration,and invasion abilities of esophageal cancer cells in a dose-dependent manner(P<0.05).Formononetin treatment significantly downregulated the immune co-stimulatory factor B7-H1 and immune escape-related factors(IL-4,IL-10,TNF-α)in a dose-dependent manner(P<0.05).Formononetin treatment significantly downregulated protein expressions of key proteins in the ERK signaling pathway(ERK1/2,c-Fos,c-Jun)in a dose-dependent manner(P<0.05).Compared with those induced with blank control,treatment of either

关 键 词：芒柄花黄素 食管肿瘤 ERK信号通路 免疫逃逸 恶性进展 

分 类 号：R735.1[医药卫生—肿瘤]