肝细胞癌（HCC）是一种常见的癌症，病死率和死亡率很高。然而，目前治疗HCC的药物有限。本研究的重点是利用高通量测序高通量筛选（HTS2）和肝癌途径相关基因签名，从中药中发现潜在的抗HCC化合物。采用HTS2鉴定草药。利用蛋白质相互作用（PPI）网络分析和计算机辅助药物设计（CADD）来鉴定关键靶点并筛选草药的候选天然产物。通过分子对接、网络药理学分析、蛋白印迹、免疫荧光染色、亚细胞分离实验、双荧光素酶报告基因实验、表面等离子共振（SPR）以及核磁共振（NMR）等方法来验证化合物与关键靶点的结合能力和抑制其功能的能力。此外，还进行了细胞存活率、集落形成、细胞周期实验和动物实验以检测化合物对HCC的抑制作用。我们对578种草药提取物对肝癌途径的84个基因表达的干扰进行了检查，并确定了前20种草药显著逆转该途径的基因表达。蛋白质相互作用网络分析确定信号转导和转录激活子3（STAT3）是肝癌途径的关键靶点之一。然后，通过使用CADD分析前20种草药中的化合物，我们发现人参皂苷F2（GF2）与STAT3具有高亲和力的结合，并通过分子对接、SPR和NMR的结果进行了进一步验证。此外，我们的结果显示GF2抑制了STAT3 Y705的磷酸化，抑制了其核转位，降低了其转录活性，并在体外和体内抑制了HCC的生长。根据这项大规模转录研究，发现了一些抗HCC的草药。GF2，一种从中药中提取的化合物，被发现是这些草药治疗HCC的化学基础。本研究还发现GF2是一种新的STAT3抑制剂，能够抑制HCC。因此，GF2代表了一种新的潜在的抗HCC治疗策略。版权所有© 2023 Elsevier GmbH。保留所有权利。

Hepatocellular carcinoma (HCC) is a common type of cancer that shows great morbidity and mortality rates. However, there are limited available drugs to treat HCC.The present work focused on discovering the potential anti-HCC compounds from traditional Chinese medicine (TCM) by employing high-throughput sequencing-based high-throughput screening (HTS2) together with the liver cancer pathway-associated gene signature.HTS2 assay was adopted for identifying herbs. Protein-protein interaction (PPI) network analysis and computer-aided drug design (CADD) were used to identify key targets and screen the candidate natural products of herbs. Molecular docking, network pharmacology analysis, western blotting, immunofluorescent staining, subcellular fractionation experiment, dual-luciferase reporter gene assay, surface plasmon resonance (SPR) as well as nuclear magnetic resonance (NMR) were performed to validate the ability of compound binding with key target and inhibiting its function. Moreover, cell viability, colony-forming, cell cycle assay and animal experiments were performed to examine the inhibitory effect of compound on HCC.We examined the perturbation of 578 herb extracts on the expression of 84 genes from the liver cancer pathway, and identified the top 20 herbs significantly reverting the gene expression of this pathway. Signal transducer and activator of transcription 3  (STAT3)  was identified as one of the key targets of the liver cancer pathway by PPI network analysis. Then, by analyzing compounds from top 20 herbs utilizing CADD, we found ginsenoside F2 (GF2) binds to STAT3 with high affinity, which was further validated by the results from molecular docking, SPR and NMR. Additionally, our results showed that GF2 suppresses the phosphorylation of Y705 of STAT3, inhibits its nuclear translocation, decreases its transcriptional activity and inhibits the growth of HCC in vitro and in vivo.Based on this large-scale transcriptional study, a number of anti-HCC herbs were identified. GF2, a compound derived from TCM, was found to be a chemical basis of these herbs in treating HCC. The present work also discovered that GF2 is a new STAT3 inhibitor, which is able to suppress HCC. As such, GF2 represents a new potential anti-HCC therapeutic strategy.Copyright © 2023 Elsevier GmbH. All rights reserved.