中华猕猴桃根抗肝癌分子调控机制研究

Molecular regulatory mechanism of Actinidia chinensis Planch root for liver cancer

目的探讨中华猕猴桃根治疗肝癌的潜在作用靶点及信号通路。方法收集癌症基因组图谱数据库(TCGA)和国际癌症基因组联合体数据库(ICGC)截至2021年12月收录的肝癌患者肿瘤组织、正常肝组织的转录组数据及临床信息。代谢相关基因来自MSigDB数据库和Gene Cards数据库。筛选中华猕猴桃根的有效活性成分及靶点。利用R 4.1.1软件获得肝癌组织中差异表达的代谢基因。将差异代谢基因与中华猕猴桃根活性成分的靶基因做交集处理,获得交集基因。交集基因进行GO功能富集分析、KEGG通路富集分析。通过Cytoscape 3.9.0软件建立“中华猕猴桃根药效成分-靶点-疾病”的网络,获取节点的度值较大的靶点。利用vina软件进行分子对接,运用PyMOL软件对结合能的绝对值较大的对接结果进行可视化。结果最终筛选出15种中华猕猴桃根有效成分化合物,涉及靶点基因326个。MSigDB数据库和Gene Cards数据库最终筛选出321个差异表达的代谢基因,与326个靶点基因进行交集映射,得到ACACA、ACSL4、AKR1C3、CYP1A2、CYP2C19等表达上调或下调的20个抗癌基因。药物活性成分-靶点网络构建结果显示,节点主要涉及槲皮素、山奈酚、β-谷甾醇等13种活性成分,对应ACACA、ACSL4、AKR1C3、CYP1A2、CYP2C19、CYP2C9等20个潜在靶点。GO富集分析结果显示,其生物学过程主要在脂肪酸代谢过程、长链脂肪酸代谢过程、不饱和脂肪酸代谢过程、烯烃化合物代谢过程、脂肪酸生物合成过程等。KEGG通路富集分析结果显示,信号通路主要涉及化学致癌作用-DNA加合物、花生四烯酸代谢、亚油酸代谢、药物代谢-细胞色素P450等。分子对接共涉及9个代谢途径,POLD1、AKR1C3、CYP2C9、ACACA共4个靶点基因,槲皮素、山奈酚2种有效成分,槲皮素-POLD1、山奈酚-AKR1C3、槲皮素-CYP2C9具有较好的结合活性。结论槲皮素和山奈酚可能是中华猕猴桃根干预肝癌细胞代谢重编过程中起主要活性的药效成分,其可能通过作用于AKR1C3、CYP2C9、ACACA、POLD1这4个靶点,参与调节肿瘤细胞中的代谢重编过程。

Objective To explore the potential therapeutic targets and signal pathways of Actinidia chinensis Planch root in the treatment of liver cancer.Methods The transcriptome data and clinical information of tumor tissues and normal liver tissues of patients with liver cancer were collected from the cancer genome atlas(TCGA)and International Cancer Genome Association Database(ICGC)up to December 2021.Metabolism related genes come from MSigDB database and Gene Cards database.The active components and targets of Actinidia chinensis Planch root were screened.The metabolic genes differentially expressed in hepatocellular carcinoma tissues were obtained by R 4.1.1 software.The differential metabolic genes were intersected with the target genes of the active components of Actinidia chinensis Planch root,and the overlapping genes were obtained.The overlapping genes were analyzed by GO functional enrichment analysis and KEGG pathway enrichment analysis.The network of Actinidia chinensis Planch root active ingredient-target-disease was established by Cytoscape 3.9.0 software,and the target with large node value was obtained.Vina software was used for molecular docking,and PyMOL software was used to visualize the docking results with large absolute value of binding energy.Results A total of 15 active components of Actinidia chinensis Planch root were screened,involving 326 target genes.A total of 321 differentially expressed metabolic genes were screened from MSigDB database and Gene Cards database,and 20 anticancer genes were obtained by cross mapping with 326 target genes,such as ACACA,ACSL4,AKR1 C3,CYP1 A2,CYP2 C19 and so on.The results of drug active component-target network construction showed that the node mainly involved 13 active components such as quercetin,kaempferol andβ-sitosterol,corresponding to 20 potential targets such as ACACA,ACSL4,AKR1 C3,CYP1 A2,CYP2 C19 and CYP2 C9.The results of GO enrichment analysis showed that the biological processes were mainly in fatty acid metabolism,long-chain fatty acid metabolism,unsaturated fatty acid metabolism,olefin metabolism,fatty acid biosynthesis and so on.The results of KEGG pathway enrichment analysis showed that the signal pathway was mainly involved in chemical carcinogenesis-DNA adducts,arachidonic acid metabolism,linoleic acid metabolism,drug metabolism-cytochrome P450 and so on.Molecular docking involves nine metabolic pathways,including POLD1,AKR1 C3,CYP2 C9 and ACACA target genes,active ingredients including quercetin and kaempferol.Quercetin-POLD1,kaempferol-AKR1 C3 and quercetin-CYP2 C9 have good binding activity.Conclusion Quercetin and kaempferol may be the main active components of Actinidia chinensis Planch root in the process of metabolic reprogramming of liver cancer cells,and they may be involved in regulating the metabolic reprogramming process of tumor cells by acting on the four targets of AKR1 C3,CYP2 C9,ACACA and POLD1.