多技术联合应用探索川陈皮素抑制骨肉瘤生长的分子机制  

Exploring the molecular mechanisms underlying Nobiletin's inhibition of osteosarcoma growth based on multi-method

作　　者：李向平 闫张若平 杨昭杰 张勇勇 陈钱 LI Xiangping;YAN-ZHANG Ruoping;YANG Zhaojie;ZHANG Yongyong;CHEN Qian(Luoyang Orthopedic Hospital of Henan Province(Orthopedic Hospital of Henan Province),Henan Zhengzhou 450000,China)

机构地区：[1]河南省洛阳正骨医院(河南省骨科医院),河南郑州450000

出　　处：《现代肿瘤医学》2025年第4期567-575,共9页Journal of Modern Oncology

基　　金：河南省中医药科学研究专项课题(编号:2022ZY1123,2022ZY1136,2023ZY2122,2024ZY3071)。

摘　　要：目的:探索川陈皮素抑制骨肉瘤生长的分子机制。方法:从GEO数据库中获取GSE70414、GSE36001和GSE14789进行数据集合并,使用R软件分析合并后的数据集获取差异表达基因,并对差异表达基因进行KEGG、GO富集分析。从swiss,TCMSP,SEA,SuperPred数据库获取川陈皮素靶点基因,取川陈皮素靶点基因和骨肉瘤差异表达基因的交集基因,并使用Cytoscape软件建立交集基因PPI网络来确定核心基因。接下来使用Lasso回归分析构建核心基因诊断模型,并通过外部数据集进行核心基因诊断模型的验证,同时使用分子对接分析川陈皮素和核心基因间的结合活性。最后使用骨肉瘤细胞系通过CCK-8、平板克隆实验评价川陈皮素对骨肉瘤细胞增殖的影响,通过RT-qPCR实验分析川陈皮素作用于骨肉瘤细胞后核心基因mRNA水平的变化。结果:差异分析成功筛选出1260个差异表达基因,其中639个上调基因,621个下调基因。KEGG信号通路包括细胞因子受体相互作用、趋化因子信号通路、癌症的转录失调等信号通路;GO功能富集分析结果涉及蛋白质结合、基因转录、激酶活性等。PPI网络的构建,识别出PIK3CG、FPR1、KDR三个核心基因;核心基因诊断模型的构建和验证结果显示三个核心基因有助于骨肉瘤患者的诊断,且其高表达与骨肉瘤患者不良预后密切相关。分子对接结果显示川陈皮素与核心基因有较好结合活性。CCK-8和平板克隆结果显示川陈皮素有效地抑制骨肉瘤细胞生长;RT-qPCR结果显示川陈皮素处理骨肉瘤细胞能显著降低PIK3CG的mRNA水平。结论:川陈皮素能显著抑制骨肉瘤细胞的生长,且川陈皮素抑制骨肉瘤细胞生长可能与PIK3CG有关。Objective:To explore the molecular mechanisms underlying Nobiletin's inhibition of OS growth.Methods:Merge GSE70414,GSE36001,and GSE14789 from the GEO database,analyze the merged dataset using R software to obtain differentially expressed genes,and perform KEGG and GO enrichment analysis on the differentially expressed genes.Retrieve the target genes of Nobiletin from the swiss,TCMSP,SEA,and SuperPred databases,and then obtain the intersection genes of Nobiletin target genes and differentially expressed genes in OS.Using Cytoscape software to establish PPI network to determine the Hub genes.Next,Lasso regression analysis will be used to construct a diagnostic model,and the Hub genes diagnostic model will be validated using an external dataset.Molecular docking analysis will also be used to determine the binding activity between Nobiletin and Hub genes.Finally,the effect of Nobiletin on the proliferation of OS cells was evaluated using CCK-8 and colony formation on OS cell lines.RT-qPCR was used to analyze the changes in mRNA levels of Hub genes after Nobiletin was applied to OS cells.Results:Differential analysis successfully screened 1260 differentially expressed genes,including 639 up-regulated genes and 621 down-regulated genes.KEGG signaling pathways include Cytokine-cytokine receptor interaction,Chemokine signaling pathway,Transcriptional misregulation in cancer,ect.GO functional enrichment analysis results involve protein binding,gene transcription,kinase activity,etc.The construction of PPI identified three hub genes,PIK3CG,FPR1,and KDR.The construction and validation of the hub genes diagnostic model showed that hub genes are helpful for the diagnosis of OS patients,and their high expression was closely related to poor prognosis in OS patients.The molecular docking results showed that nobiletin has good binding activity with hub genes.CCK-8 and colony formation capacity results showed that nobiletin inhibited the growth of OS cells.RT-qPCR results showed that treatment of OS cells with nobiletin significa

关 键 词：川陈皮素 骨肉瘤 PIK3CG 

分 类 号：R738.1[医药卫生—肿瘤]