Jianpi Jiedu Xiaozheng Fang Regulates Hepatocellular Carcinoma Proliferation and Metastasis Based on Network Pharmacology

Hepatocellular carcinoma (HCC) is a primary malignant tumour that impacts patients' quality of life. Currently, clinical experience from The First Affiliated Hospital of Guangzhou University of Chinese Medicine suggests that Jianpi Jiedu Xiaozheng Fang (JPJDXZF) demonstrates promising efficacy in the treatment of HCC. We aimed to explore the mechanisms of JPJDXZF in HCC based on network pharmacology. The components and their relevant targets of JPJDXZF were identified using databases such as SymMap, TCMID, TCMSP, and TCM-ID. Following ADME screening, 1443 active components of JPJDXZF were identified, and 435 corresponding drug targets were predicted using the SwissTargetPrediction database. Subsequently, prognosis-related differentially expressed genes (DEGs) associated with HCC were analyzed using TCGA and GTEx datasets, and a gene expression matrix was derived. Key genes involved in HCC regulation were identified, and functional analyses were performed. Furthermore, we explored the regulatory effects of JPJDXZF at the cellular, organoid, and animal levels. We identified 18 intersecting genes between HCC prognosis-related genes and JPJDXZF-target genes. Venn diagram analysis successfully identified BIRC5 and CYP2E1 as two potential targets for JPJDXZF in treating HCC. Pathway enrichment analysis indicated that the core targets of JPJDXZF were enriched in multiple signalling pathways, including the Hippo pathway, in which BIRC5 is involved as a downstream regulatory gene. In in vitro experiments, JPJDXZF-containing serum significantly reduced the viability and migration of HepG2 and MHCC97-H cells, leading to a decrease in organoid diameter and ATP activity in HCC organoids. In in vivo experiments, tumours in nude mice treated with JPJDXZF exhibited reduced volume and weight, along with decreased expression of BIRC5 and Hippo pathway effectors YAP and TAZ. At the mechanistic level, JPJDXZF treatment was associated with altered Hippo pathway-related signalling, accompanied by reduced YAP/TAZ activity and changes in BIRC5 expression, together with effects on HCC cell proliferation and apoptosis. In addition, siMST1/2 interference and EMT inhibitor-1 treatment partially attenuated the effects of JPJDXZF on cell viability, migration, and apoptosis. JPJDXZF regulates BIRC5 expression in association with Hippo pathway activity in HCC. In vitro, in vivo, and molecular mechanism analyses support JPJDXZF as a potential therapeutic strategy for HCC by modulating key proteins in the Hippo pathway, thus affecting HCC cell proliferation, apoptosis, and migration.