葛根素靶向线粒体抑制乳腺癌细胞增殖和侵袭的机制  

Mechanism of puerarin targeting the mitochondria to inhibit proliferation and invasion of breast cancer cells

作　　者：余才翰 李岱[1] 谢敏 Yu Caihan;Li Dai;Xie Min(School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning 437100;Dept of Ophthalmology,Xianning Central Hospital,The First Affiliated Hospital of Hubei University of Science and Technology,Xianning 437100)

机构地区：[1]湖北科技学院医学部药学院,咸宁437100 [2]咸宁市中心医院(湖北科技学院第一附属医院)眼科,咸宁437100

出　　处：《安徽医科大学学报》2024年第4期627-633,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81901149);湖北省自然科学基金(编号:2022CFB356)。

摘　　要：目的利用网络药理学、分子对接及体外实验验证探讨葛根素对人乳腺癌细胞HCC1806增殖、侵袭和凋亡的作用及机制。方法GEO数据库收集乳腺癌和线粒体疾病数据,GEO2R分析差异性表达基因;韦恩图分析乳腺癌与线粒体数据集的重叠基因;GO和KEGG功能富集分析重叠基因;STRING和Cytoscape分析重叠基因相互作用网络并筛选核心基因;Autodock对接葛根素和核心基因;通过CCK-8、EdU、TUNEL和Transwell实验检测细胞增殖、凋亡和侵袭能力,采用Mito-Tracker实验检测线粒体膜电位,Western blot检测蛋白质表达水平。体内实验通过乳腺癌皮下异种移植和免疫组化分析葛根素的抗肿瘤药效。结果乳腺癌和线粒体疾病共有132个重叠基因,筛选出10个核心差异性表达基因,其中双丝氨酸/苏氨酸和酪氨酸蛋白激酶(DST)在乳腺癌组织中呈低表达。葛根素可结合并上调DST表达,抑制人乳腺癌细胞HCC1806增殖和侵袭,促进细胞凋亡,增加凋亡相关蛋白Cleaved-Caspase 3和Bax表达,下调抗凋亡蛋白Bcl-2的表达,降低线粒体膜电位。体内实验显示葛根素可抑制乳腺癌的生长,上调瘤体组织中DST的表达。结论葛根素抑制乳腺癌细胞增殖及侵袭并促进癌细胞凋亡进而抑制乳腺癌生长。Objective To investigate the effect and mechanism of puerarin on the proliferation,invasion and apoptosis of breast cancer cell HCC1806 via network pharmacology,molecular docking and in vitro experiments.Methods The data of breast cancer and mitochondrial diseases were collected from GEO database,and the differentially expressed genes were analyzed by GEO2R.Overlapping genes between breast cancer and mitochondrial database were analyzed by Venn diagram.GO and KEGG enrichment analyzed the overlapping genes.STRING and Cytoscape analyzed overlapping gene interaction networks and screen core genes.Interaction between puerarin and core genes was docked with autodock.Cell proliferation,apoptosis and invasion capacity were measured by CCK-8,EdU,TUNEL and Transwell experiments,mitochondrial membrane potential was measured by Mito-Tracker experiments and protein expression levels were measured by Western blot.Anti-tumor efficacy of puerarin was analyzed by subcutaneous xenograft of breast cancer and immunohistochemical assay in vivo.Results Among 132 overlapping genes in breast cancer and mitochondrial disease,10 core differentially expressed genes were selected.Among these core genes,dual serine/threonine and tyrosine protein kinase(DST)was low expressed in breast cancer tissues.Puerarin bound to DST,up-regulated its expression,inhibited the proliferation and invasion of HCC1806 breast cancer cells,promoted breast cell apoptosis,increased the expression levels of apoptosis-related proteins Cleaved-Caspase 3 and Bax,down-regulated the expression of anti-apoptosis protein Bcl-2,and decreased mitochondrial membrane potential.In vivo,puerarin inhibited the growth of breast cancer and up-regulated the expression of DST in tumor tissues.Conclusion Puerarin inhibits the proliferation and invasion of breast cancer cells,promotes the apoptosis of cancer cells and inhibits breast cancer growth.

关 键 词：葛根素 线粒体 乳腺癌 网络药理学 分子对接 DST 

分 类 号：R737.9[医药卫生—肿瘤]