白桦脂酸对乳腺癌的干预作用及核心通路预测  

Intervention Effect of Betulinic Acid on Breast Cancer and Core Pathway Prediction

作　　者：倪雯婷 邢中夫 孙成宏 姚景春 张晓平[1] 张贵民 NI Wenting;XING Zhongfu;SUN Chenghong;YAO Jingchun;ZHANG Xiaoping;ZHANG Guimin(Shandong University of Traditional Chinese Medicine,Jinan 250355;State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 276006)

机构地区：[1]山东中医药大学,济南250355 [2]鲁南制药集团股份有限公司经方与现代中药融合创新全国重点实验室,临沂276006

出　　处：《中药药理与临床》2024年第6期50-58,共9页Pharmacology and Clinics of Chinese Materia Medica

基　　金：山东省自然基金创新发展联合基金(编号:ZR2022LZY029)。

摘　　要：目的:探索白桦脂酸治疗乳腺癌的潜在作用机制。方法:分别通过PharmMapper、Genecards、Drugbank和Therapeutic Target Database (TTD)数据库获得药物和疾病靶点,用String数据库构建蛋白之间的相互作用关系,通过Metascape数据库获得基因本体(GO)和基因组百科全书(KEGG)通路富集分析。用Autodock vina软件进行分子对接并使用Pymol对其进行可视化分析。使用Kaplan-Meier Plotter对关键核心靶点进行预后生存分析。采用四甲基偶氮唑蓝(MTT)法、克隆形成试验、细胞划痕试验、Transwell试验观察白桦脂酸对乳腺癌MCF-7细胞增殖和迁移能力的影响;AO/EB染色及Annexin V-FITC/PI染色观察白桦脂酸对细胞凋亡的影响;Western blot法观察白桦脂酸对PI3K/AKT/mTOR信号通路的影响。结果:筛选出白桦脂酸与乳腺癌的交集靶点的蛋白相互作用网络共有33个节点和100条边,平均节点度值为6.06,筛选得到的关键核心靶点为NR3C1、SRC、ALB、MAPK8、PGR和GRB2,其与白桦脂酸对接得到的结合能均<-5 KJ/mol;GO功能富集中的生物学过程得到473个条目,主要有细胞对激素刺激的反应、细胞内受体信号通路、介导激素信号通路等,分子功能得到42个条目,主要有脂质结合、蛋白激酶活性、转录因子结合等,细胞组分得到7个条目,主要涉及受体复合物、转录因子复合物、膜阀等;KEGG富集分析中共得到172条信号通路,主要有PI3K/AKT通路、Ras通路、ErbB通路、GnRH通路和EGFR通路等。体外实验验证显示:白桦脂酸可以抑制乳腺癌细胞的增殖,且具有剂量依赖性和时间依赖性;白桦脂酸可抑制乳腺癌细胞的迁移、侵袭能力;从定性、定量两个方面证实白桦脂酸可以促进细胞凋亡;白桦脂酸可以显著下调PI3K、AKT和mTOR蛋白表达(P<0.01)。结论:白桦脂酸可抑制乳腺癌细胞MCF-7的增殖、迁移等恶性生物学行为,其机制可能与抑制PI3K/AKT/mTOR信号通路蛋白表达相关。Objective:To explore the potential mechanism of betulinic acid in the treatment of breast cancer.Methods:Drug and disease targets were obtained through PharmMapper,Genecards,Drugbank and Therapeutic Target Database(TTD).The protein-protein interaction was revealed in the String.Metascape was used for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Molecular docking was carried out by Autodockvina and visual analysis was carried out by Pymol.Kaplan-MeierPlotter was employed to analyze the prognosis and survival of the key targets.The effect of betulinic acid on the proliferation and migration of breast cancer cells was detected by MTT method,clone formation assay,wound healing assay and Transwell assay.AO/EB staining and Annexin V-FITC/PI staining were performed to observe the effect of betulinic acid on cell apoptosis,while its effect on PI3K/AKT/mTOR signaling pathway was detected by Western blot.Results:From the protein-protein interaction network of common targets of betulinic acid and breast cancer,33 nodes and 100 edges were selected,with an average node degree of 6.06.The core targets selected were NR3C1,SRC,ALB,MAPK8,PGR and GRB2.The binding energy of betulinic acid with the targets were all less than癬5 kcal/mol.There were 473 entries observed in the biological process of GO function enrichment,mainly including cellular response to hormone stimulus,intracellular receptor signaling pathway,and hormone-mediated signaling pathway,42 entries in molecular function,mainly including lipid binding,protein kinase activity,and transcription factor binding,and 7 entries in cellular components,mainly involving receptor complexes,transcription factor complexes,membrane raft,etc.A total of 172 signaling pathways were obtained from KEGG enrichment analysis,including PI3K/AKT,Ras,ErbB,GnRH and EGFR signaling pathways.In vitro experiments showed that betulinic acid inhibited the proliferation of breast cancer cells in a dose-and time-dependent manner,as well as the invasion and migra

关 键 词：白桦脂酸 网络药理学 乳腺癌 磷脂酰肌醇激酶 蛋白激酶 雷帕霉素 

分 类 号：R285[医药卫生—中药学]