摘要
目的:探究加味旋覆代赭汤(XFDZ)治疗食管鳞癌(ESCC)的潜在分子机制。方法:通过中药系统药理学数据库与分析平台(TCMSP)、中药分子机制的生物信息学分析数据库(BATMAN-TCM)筛选XFDZ的有效成分及作用靶点,同时基于基因表达综合数据库(GEO)、人类基因综合数据库(GeneCards)检索ESCC关键基因,将二者整合,结果通过Cytoscape可视化,得到XFDZ治疗ESCC的潜在靶点,对预测的靶点进行富集分析。采用人ESCC KYSE150细胞系构建小鼠皮下移植瘤模型,对XFDZ进行抑癌效果及安全性评估。运用实时定量逆转录聚合酶链式反应(RT-qPCR)和蛋白质免疫印迹(Western blot)实验在小鼠皮下瘤组织中对预测的关键靶点进行验证。结果:基于网络药理学结合分子生物学研究表明,XFDZ通过多靶点、多通路调控方式抑制ESCC进展,其关键作用机制与阻断氧化应激介导的磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路激活密切相关。主要作用靶点包括PI3K、Akt、磷酸酶和张力蛋白同系物(PTEN)等。XFDZ能有效抑制人ESCC细胞KYSE150构建的小鼠皮下移植瘤生长,表现为瘤体积及质量明显降低(P<0.05),且无明显血液毒性及肝肾损伤。分子生物学实验证实,XFDZ干预可显著下调小鼠皮下瘤中增殖细胞核抗原(PCNA)的蛋白及mRNA表达(P<0.001,P<0.01),下调网络药理学预测的关键靶点PI3K、Akt的蛋白磷酸化水平(P<0.001,P<0.05),并上调PTEN的蛋白及mRNA表达水平(P<0.001,P<0.05)。结论:XFDZ抑制ESCC的作用涉及多靶点、多通路,其中PI3K/Akt信号通路可能尤为关键。
Objective:To investigate the potential molecular mechanisms by which modified Xuanfu Daizhe Decoction(XFDZ)treats esophageal squamous cell carcinoma(ESCC).Methods:Effective components and their corresponding action targets were identified via Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine(BATMAN-TCM).Concurrently,key genes associated with ESCC were extracted from Gene Expression Omnibus(GEO)and GeneCards databases.The integration of these datasets was visualized using Cytoscape to elucidate the potential targets of XFDZ in the treatment of ESCC.Pathway enrichment analyses were performed on the selected targets.The mouse subcutaneous tumor model with a human ESCC cell line,KYSE150 cells was employed to assess the anti-cancer efficacy and safety profile of XFDZ.The expression levels of predicted key targets in tumor tissues of mice were quantified via quantitative real-time reverse transcription polymerase chain reaction(RT-qPCR)and Western blot analysis.Results:Integrated network pharmacology and molecular biology studies revealed that XFDZ inhibits ESCC progression via multi-target and multi-pathway regulatory mechanisms.The key mechanism involves suppressing oxidative stress-mediated activation of the phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway,with core targets including PI3K,Akt,and phosphatase and tensin homolog(PTEN).In a KYSE150-derived subcutaneous xenograft mouse model,XFDZ significantly attenuated tumor growth(P<0.05),manifested by reduced tumor volume and mass,while exhibiting no overt hematotoxicity or hepatorenal toxicity.The anti-tumor mechanism of XFDZ was confirmed by molecular experiments,showing marked downregulation of proliferating cell nuclear antigen(PCNA)protein and mRNA expression(P<0.001,P<0.01),suppression of PI3K and Akt phosphorylation(P<0.001,P<0.05),and upregulation of PTEN protein and mRNA expression in subcutaneous tumor tissues(P<0.001,P<0.05).Conclusion:XFDZ exerts its inhibitory effects on ESCC through multi-target and multipathway mechanisms,with the PI3K/Akt signaling pathway playing a pivotal role.
作者
郑苗苗
王聪聪
郝苗秀
金星
陈文连
ZHENG Miaomiao;WANG Congcong;HAO Miaoxiu;JIN Xing;CHEN Wenlian(Cancer Institute,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Shanghai Frontiers Science Center of Disease and Syndrome Biology of Inflammatory Cancer Transformation,Shanghai 200032,China)
出处
《上海中医药大学学报》
2025年第3期43-53,共11页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
国家重点研发计划项目(2023YFC3503201,2022YFC3500202)
国家自然科学基金资助项目(32170778,82304780)。
关键词
加味旋覆代赭汤
食管鳞癌
网络药理学
皮下移植瘤
氧化应激
PI3K/AKT信号通路
modified Xuanfu Daizhe Decoction
esophageal squamous cell carcinoma
network pharmacology
subcutaneous xenograft tumor
oxidative stress
PI3K/Akt signaling pathway
作者简介
郑苗苗,女,在读硕士生,主要从事食管癌代谢的中西医结合基础与转化研究;通信作者:陈文连,研究员,博士生导师,E-mail:chenwl8412@shutcm.edu.cn。
