Zhu Mingyang, He Qingmin, Wang Yanan, Duan Liying, Rong Kang, Wu Yingying, Ding Ye, Mi Yang, Ge Xiaoyang, Yang Xiaocui, Yu Yong
Department of Gastroenterology, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Henan Key Laboratory of Helicobacter Pylori & Microbiota and Gastrointestinal Cancer, Marshall B. J. Medical Research Center of Zhengzhou University, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Front Pharmacol. 2023 Oct 12;14:1238841. doi: 10.3389/fphar.2023.1238841. eCollection 2023.
Aloe-emodin (AE) is an anthraquinone compound extracted from the rhizome of the natural plant rhubarb. Initially, it was shown that AE exerts an anti-inflammatory effect. Further studies revealed its antitumor activity against various types of cancer. However, the mechanisms underlying these properties remain unclear. Based on network pharmacology and molecular docking, this study investigated the molecular mechanism of AE in the treatment of hepatocellular carcinoma (HCC), and evaluated its therapeutic effect through experiments. CTD, Pharmmapper, SuperPred and TargetNet were the databases to obtain potential drug-related targets. DisGenet, GeneCards, OMIM and TTD were used to identify potential disease-related targets. Intersection genes for drugs and diseases were obtained through the Venn diagram. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of intersecting genes were conducted by the website of Bioinformatics. Intersection genes were introduced into STRING to construct a protein-protein interaction network, while the Cytoscape3.9.1 software was used to visualize and analyze the core targets. AutoDock4.2.6 was utilized to achieve molecular docking between drug and core targets. experiments investigated the therapeutic effects and related mechanisms of AE. 63 overlapped genes were obtained and GO analysis generated 3,646 entries by these 63 intersecting genes. KEGG analysis mainly involved apoptosis, proteoglycans in cancer, TNF signaling pathway, TP53 signaling pathway, PI3K-AKT signaling pathway, etc. AKT1, EGFR, ESR1, TP53, and SRC have been identified as core targets because the binding energies of them between aloe-emodin were less than -5 kcal/Mol.The mRNA and protein expression, prognosis, mutation status, and immune infiltration related to core targets were further revealed. The involvement of AKT1 and EGFR, as well as the key target of the PI3K-AKT signaling pathway, indicated the importance of this signaling pathway in the treatment of HCC using AE. The results of the Cell Counting Kit-8 assay and flow analysis demonstrated the therapeutic effect of AE. The downregulation of EGFR, PI3KR1, AKT1, and BCL2 in mRNA expression and PI3KR1, AKT,p-AKT in protein expression confirmed our hypothesis. Based on network pharmacology and molecular docking, our study initially showed that AE exerted a therapeutic effect on HCC by modulating multiple signaling pathways. Various analyses confirmed the antiproliferative activity and pro-apoptotic effect of AE on HCC through the PI3K-AKT signaling pathway. This study revealed the therapeutic mechanism of AE in the treatment of HCC through a novel approach, providing a theoretical basis for the clinical application of AE.
芦荟大黄素（AE）是从天然植物大黄的根茎中提取的一种蒽醌化合物。最初，研究表明AE具有抗炎作用。进一步研究发现其对多种类型癌症具有抗肿瘤活性。然而，这些特性背后的机制仍不清楚。基于网络药理学和分子对接，本研究探讨了AE治疗肝细胞癌（HCC）的分子机制，并通过实验评估其治疗效果。CTD、Pharmmapper、SuperPred和TargetNet是用于获取潜在药物相关靶点的数据库。DisGenet、GeneCards、OMIM和TTD用于识别潜在疾病相关靶点。通过维恩图获得药物和疾病的交集基因。利用生物信息学网站对交集基因进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析。将交集基因导入STRING构建蛋白质-蛋白质相互作用网络，同时使用Cytoscape3.9.1软件对核心靶点进行可视化和分析。利用AutoDock4.2.6实现药物与核心靶点之间的分子对接。实验研究了AE的治疗效果及相关机制。获得了63个重叠基因，GO分析由这63个交集基因产生了3646个条目。KEGG分析主要涉及细胞凋亡、癌症中的蛋白聚糖、TNF信号通路、TP53信号通路、PI3K-AKT信号通路等。AKT1、EGFR、ESR1、TP53和SRC已被确定为核心靶点，因为它们与芦荟大黄素之间的结合能小于-5千卡/摩尔。进一步揭示了与核心靶点相关的mRNA和蛋白表达、预后、突变状态及免疫浸润情况。AKT1和EGFR的参与以及PI3K-AKT信号通路的关键靶点表明该信号通路在AE治疗HCC中的重要性。细胞计数试剂盒-8检测和流式分析结果证明了AE的治疗效果。EGFR、PI3KR1、AKT1和BCL2的mRNA表达下调以及PI3KR1、AKT、p-AKT的蛋白表达下调证实了我们的假设。基于网络药理学和分子对接，我们的研究初步表明AE通过调节多种信号通路对HCC发挥治疗作用。各种分析证实了AE通过PI3K-AKT信号通路对HCC具有抗增殖活性和促凋亡作用。本研究通过一种新方法揭示了AE治疗HCC的机制，为AE的临床应用提供了理论依据。
