Investigation on the Inhibitory Mechanism of Chai Hu Hua Ji Tang on Liver Cancer Based on a Network Pharmacology Analysis.

Context: Hepatocellular carcinoma (HCC) poses a serious threat to human health due to its high incidence and mortality rates. In recent years, the application of traditional Chinese medicine (TCM) in the comprehensive treatment of liver cancer has gained increasing attention. Clinical practices have demonstrated the mighty efficacy of Chai Hu Hua Ji Tang (CHHJT) in liver cancer treatment, yet its underlying mechanism remains unexplored. Objective: This article reveals the underlying of CHHJT in the context of hepatic malignancies. Materials and Methods: CHHJT and HCC-related targets were screened through network pharmacology. PLC/PRF/5 and MHCC97L cells were treated with CHHJT as the experimental group, and those without CHHJT as the control group. After 24 and 48 h, IC₅₀, cloning, scratch, invasion, and Western blot/quantitative real-time PCR were assayed. Subsequently, a model of orthotopic liver cancer was developed in BALB/c nude mice. A volume of 0.2 mL of PBS served as the control group, while the experimental group received 0.2 mL of a 3.55 g/mL CHHJT solution. After 4 weeks, the in vivo effect of CHHJT was evaluated. Results: According to our results of IC₅₀, a concentration of 1600 μg/mL was the most effective dosage for CHHJT to suppress growth of HCC cells. Additionally, CHHJT was found to curb cellular proliferation, migration, and invasiveness. Transcription and protein levels of FLT3, PIK3CA, and AKT1 targets reduced following CHHJT treatment. In the nude mouse model, CHHJT treatment greatly reduced tumor cell volume, integrated tumor cell structure, markedly reduced nodules, well-grown cells, and substantially increased apoptosis. Discussion and Conclusion: CHHJT is likely to impede the PI3K/AKT signaling pathway by downregulating FLT3 protein expression, ending up with suppression of HCC cell differentiation and proliferation, as well as their viability. Future research could stratify patients based on their likelihood of responding to PI3K/AKT pathway-targeted therapies. This could aid in patient selection and optimize treatment outcomes.