摘要
目的:揭示芒柄花黄素对肝癌免疫逃逸的作用机制。方法:采用不同浓度(50、100、200μg/ml)芒柄花黄素处理人肝癌细胞系HepG2 24 h,CCK-8检测细胞增殖,Annexin V-FITC/PI试剂盒检测细胞凋亡。雄性C57BL/6小鼠皮下接种HepG2细胞(4×105个)建立荷瘤小鼠模型,10/50 mg(/kg·d)芒柄花黄素治疗28 d,计算抑瘤率。qRT-PCR、Western blot检测小鼠肝癌组织和HepG2细胞TLR4和NF-κB p65表达,ELISA检测小鼠血清和HepG2细胞TNF-α、IL-1β和IL-10水平。结果:芒柄花黄素可提高HepG2细胞增殖抑制率和凋亡率(P<0.05),降低荷瘤小鼠血清和HepG2细胞TNF-α、IL-1β和IL-10水平(P<0.05),提高荷瘤小鼠抑瘤率(P<0.05),降低荷瘤小鼠肿瘤组织和HepG2细胞TLR4和细胞核NF-κB p65蛋白表达(P<0.05),且呈剂量依赖性。结论:芒柄花黄素通过抑制TLR4/NF-κB通路介导的肿瘤免疫逃逸抑制肝癌发生发展。
Objective:To reveal mechanism of formononetin on immune escape of liver cancer.Methods:Human hepatoma cell line HepG2 were treated with formononetin at different concentrations(50,100,200μg/ml)for 24 h,cell proliferation was detected by CCK-8,and cell apoptosis was detected by Annexin V-FITC/PI kit.Male C57BL/6 mice were subcutaneously inoculated with HepG2 cells(4×105 cells)to establish a tumor-bearing mouse model,and then treated with 10 or 50 mg(/kg·d)formononetin for 28 d,tumor inhibition rate was calculated.TLR4 and NF-κB p65 expressions in mouse liver cancer tissues and HepG2 cells were detected by qRT-PCR and Western blot.TNF-α,IL-1βand IL-10 levels in mouse serum and HepG2 cell culture supernatant were detected by ELISA.Results:Formononetin increased proliferation inhibition rate and apoptosis rate of HepG2 cells(P<0.05),reduced levels of TNF-α,IL-1βand IL-10 in serum of tumor-bearing mice and HepG2 cells(P<0.05),increased tumor inhibition rate of tumor-bearing mice(P<0.05),decreased TLR4 and nuclear NF-κB p65 protein expressions in tumor tissues of tumor-bearing mice and HepG2 cells(P<0.05),which were in a dose-dependent manner.Conclusion:Formononetin inhibits occurrence and development of liver cancer by inhibiting tumor immune escape mediated by TLR4/NF-κB pathway.
作者
李汨
蒋承志
陈建婷
王军艳
LI Mi;JIANG Chengzhi;CHEN Jianting;WANG Junyan(Department of Physiology,Basic Medical College,Shaanxi University of Traditional Chinese Medicine,Xi'an 712046,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2023年第9期1872-1877,共6页
Chinese Journal of Immunology
