解放军医学杂志2025，Vol.50Issue(5)：581-591,11.DOI:10.11855/j.issn.0577-7402.0351.2025.0126
基于网络药理学及体外实验探讨香蒲新苷对肺腺癌的作用及其分子机制
Exploring the effects and underlying mechanisms of typhaneoside on lung adenocarcinoma based on network pharmacology and in vitro experiments
摘要
Abstract
Objective To investigate the effects and underlying molecular mechanisms of typhaneoside(TYP)on lung adenocarcinoma based on network pharmacology and in vitro experiments.Methods TYP-lung adenocarcinoma-related genes were obtained from the prediction target website SwissTargetPrediction,The Cancer Genome Atlas(TCGA)database,GeneCards database,and Gene Expression Omnibus(GEO)database.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed on the obtained TYP-lung adenocarcinoma-related genes.Protein-protein network interactions analysis was carried out using the String database,and molecular docking was conducted with the Vina software.These network pharmacological analysis methods were employed to explore the theoretical action pathways of TYP on lung adenocarcinoma.PC9 cells were divided into control group(normal culture),low-TYP group(treated with 50 μmol/L TYP for 48 h),high-TYP group(treated with 100 μmol/L TYP for 48 h),and ferrostatin-1(Fer-1)+TYP group(treated with 1 μmol/L Fer-1+100 μmol/L TYP for 48 h).The cell viability was detected by the CCK-8 assay,the cell proliferation ability was detected by the EdU assay,and the cell migration and invasion ability was detected by scratch assay,Transwell migration assay and Transwell invasion assay.The occurrence of ferroptosis was detected by reduced glutathione(GSH)assay kit,reactive oxygen species(ROS)assay kit,mitochondrial membrane potential assay kit,ferrous ion fluorescent probe,and transmission electron microscopy.Protein and mRNA expression levels of ferroptosis-related key molecules,solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),were measured by Western blotting and reverse transcription-quantitative PCR(RT-qPCR).PC9 cells were transfected with SLC7A11 overexpression plasmid and divided into vector group(transfected with empty plasmid),vector+TYP group(transfected with empty plasmid and treated with 100 μmol/L TYP for 48 h),OE-SLC7A11 group(transfected with SLC7A11 overexpression plasmid)and OE-SLC7A11+TYP group(transfected with SLC7A11 overexpression plasmid and treated with 100 μmol/L TYP for 48 h).The molecular mechanism of SLC7A11 in the effect of TYP on PC9 was verified by CCK-8 assay,ferrous ion fluorescence probe,and protein expression levels of SLC7A11 and GPX4.Results A total of 73 TYP-lung adenocarcinoma-related target genes were predicted.GO analysis showed that the target genes were mainly involved in biological processes such as positive regulation of kinase activity,and were enriched in cellular components like transferase complex(transferring phosphorus-containing groups),revealing molecular functions such as transmembrane receptor protein tyrosine kinase activity.KEGG analysis identified 124 related pathways,mainly including epidermal growth factor receptor(EGFR)tyrosine kinase inhibitor resistance pathway.Protein-protein interaction network analysis obtained 7 core targets such as serine/threonine kinase 1(Akt1).Molecular docking results showed that the binding energies of TYP with core target genes and the ferroptosis-related proteins(SLC7A11 and GPX4)were all≤-5.0 kcal/mol,indicating significant binding activity.CCK-8 assay showed that TYP significantly inhibited PC9 cell viability(P<0.05).EdU assay demonstrated that the proportion of proliferating cells in TYP group was significantly lower than that in control group(P<0.05).Scratch assay,Transwell migration assay,and invasion assay revealed that,compared with control group,the migration and invasion abilities of PC9 cells in TYP group significantly decreased(P<0.05),and GSH content and mitochondrial membrane potential in TYP group also significantly decreased(P<0.05),while ROS and Fe2+contents increased significantly(P<0.01),and protein and mRNA expression levels of SLC7A11 and GPX4 decreased significantly(P<0.05).Transmission electron microscopy results showed that cells in TYP group exhibited specific ferroptosis-related changes such as reduced mitochondrial cristae and increased membrane density,compared with control group.Compared with vector+TYP group,OE-SLC7A11+TYP group had higher cell viability(P<0.05),lower Fe2+content(P<0.05),and higher protein expression levels of SLC7A11 and GPX4(P<0.05).Conclusion TYP may induce ferroptosis in lung adenocarcinoma cells and inhibit their malignant behaviors such as proliferation,migration and invasion by regulating the SLC7A11-GPX4 axis.关键词
铁死亡/香蒲新苷/肺腺癌/网络药理/分子对接Key words
ferroptosis/typhaneoside/lung adenocarcinoma/network pharmacology/molecular docking分类
医药卫生引用本文复制引用
刘梦茹,阎雪..基于网络药理学及体外实验探讨香蒲新苷对肺腺癌的作用及其分子机制[J].解放军医学杂志,2025,50(5):581-591,11.基金项目
辽宁省教育厅科学技术研究项目(JYTJCZR2020057) This work was supported by the Science and Technology Research Project of Education Department of Liaoning Province(JYTJCZR2020057) （JYTJCZR2020057）
