Anti-Hepatocellular Carcinoma Effect of Jianpi Huayu Formula: Revealing the Molecular Mechanisms of the CXCR4 - Smad2/3 Pathway in Regulating Vasculogenic Mimicry Formation and Epithelial-Mesenchymal Transition.

Hepatocellular carcinoma (HCC) is a malignant tumor with high global incidence and mortality rates. Jianpi Huayu Formula (JPHY) has demonstrated therapeutic potential in HCC, but its underlying mechanisms remain unclear. This study aimed to investigate the anti-HCC effects of JPHY and its molecular mechanisms, focusing on its regulatory roles in vasculogenic mimicry (VM) formation and epithelial-mesenchymal transition (EMT) progression. Sk-Hep-1 cells were treated with 5%, 10%, or 20% JPHY-medicated serum (JPHY-MS). Cell proliferation, migration, VM, and EMT were assessed using CCK-8, wound healing, tube formation, and Western blot assays. Molecular docking was employed to analyze the binding between core components of JPHY and CXCR4. A stable CXCR4-overexpressing cell line was established, and the regulatory effect of CXCR4 on the Smad2/3 pathway was observed following intervention with the Smad2/3-specific inhibitor SB431542. Finally, a retrospective study was conducted using JPHY-MS intervention. In vivo experiments: Mice bearing HCCLM3 xenografts were divided into control, model, low-dose (6.24 g/kg), and high-dose (24.96 g/kg) JPHY groups. Tumor size and body weight were measured every 3 days. The effects of drug intervention on the CXCR4-Smad2/3 pathway, EMT, and VM were examined via immunohistochemistry (IHC), CD31/PAS double staining, and Western blot analysis. JPHY may impede HCC progression and metastasis through CXCR4 downregulation-mediated suppression of Smad2/3 signaling pathway activation, concomitantly modulating VM formation and EMT processes. JPHY suppresses HCC progression by targeting the CXCR4-Smad2/3 pathway, highlighting its potential as a novel HCC therapeutic.