Tripterygium wilfordii Hook F (TwHF) exhibits anti-tumor efficacy in pancreatic ductal adenocarcinoma (PDAC), however the pharmacological mechanisms are unclear due to complicated formulae and target genes. Using Traditional Chinese Medicine Systems Pharmacology and GeneCards databases, we performed a network pharmacology (NP) of TwHF and screened out 22 ingredients and 25 target genes associated with PDAC. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the 25 target genes were performed. Using STRING database, protein-protein interaction network of the 25 target genes was constructed, and indicated that triptolide (TL)-plasminogen activator urokinase (PLAU) as a potential target for PDAC treatment. Hence, in vitro experiments were performed and validated that TL inhibited PDAC cell proliferation and migration by suppressing PLAU expression. The results of Western blot suggested that PLAU activated endothelial-mesenchymal transition (EMT) progression. In two Gene Expression Omnibus datasets (GSE16515 and GSE28735), PLAU was up-regulated in tumor tissues, and PLAU overexpression was associated with poor overall survival of PDAC cohort of The Cancer Genome Atlas (P < 0.01). Immunohistochemistry illustrated that overexpression of PLAU protein was related to lymph node metastasis in 20 PDAC patients (P < 0.01). Based on NP of TwHF, we identified and validated that TL-PLAU could serve as a potential target for PDAC treatment.

雷公藤雷公藤钩F（TwHF）在胰腺导管腺癌（PDAC）中表现出抗肿瘤功效，但是由于复杂的配方和目标基因，其药理机制尚不清楚。使用中药系统药理学和GeneCards数据库，我们进行了TwHF的网络药理学（NP），并筛选了22种与PDAC相关的成分和25个靶基因。对25个靶基因进行了基因本体论和《京都议定书全书》。使用STRING数据库，构建了25个靶基因的蛋白质-蛋白质相互作用网络，并表明雷公藤内酯醇（TL）-纤溶酶原激活物尿激酶（PLAU）作为PDAC治疗的潜在靶标。因此，进行了体外实验，并验证了TL通过抑制PLAU表达来抑制PDAC细胞增殖和迁移。蛋白质印迹的结果表明，PLAU激活了内皮-间质转化（EMT）进程。在两个基因表达综合数据集（GSE16515和GSE28735）中，肿瘤组织中的PLAU上调，而PLAU的过表达与《癌症基因组图集》的PDAC队列总体存活率低有关（P  <0.01）。免疫组织化学表明，20例PDAC患者的PLAU蛋白过表达与淋巴结转移有关（P  <0.01）。基于TwHF的NP，我们确定并验证了TL-PLAU可以作为PDAC治疗的潜在靶标。