Zuojin capsule improves T cell exhaustion and tumor immune microenvironment of hepatocellular carcinoma through the mTOR-eIF4E/p70S6K-CDK1 pathway.

Background & Aims: Hepatocellular carcinoma (HCC) is a major health concern. T cell exhaustion (Tex), a state of T cell dysfunction characterized by reduced effector function and increased expression of inhibitory receptors. This study aimed to explore the mechanism by which Zuojin capsule (ZJC, Methods: To identify HCC-related and Tex-associated targets, two HCC expression microarray datasets were integrated. Targets related to Tex were retrieved from GeneCards and Online Mendelian Inheritance in Man (OMIM) databases. Active compounds in ZJC were screened. A protein-protein interaction (PPI) network of overlapping targets was constructed using STRING to identify core functional modules. To verify the anti-proliferative effect of ZJC on HCC cells, the CCK-8 assay was performed to detect the viability of Hep3B and HepG2.2.15 cells treated with gradient concentrations of ZJC. Western Blot analysis was conducted to measure the protein expression levels of key molecules. Immunohistochemical (IHC) staining was used to assess the proliferation index of tumor cells, the infiltration of immune cells, and the expression of immune-related markers. Results: HCC-related genes, Tex targets, and ZJC targets were identified through bioinformatics analysis, 136 overlapping targets were obtained. ZJC inhibited Hep3B/HepG2.2.15 cell proliferation with IC Conclusion: ZJC exerts dual anti-tumor effects by inhibiting the mTOR-eIF4E/p70S6K-CDK1 pathway and remodeling the immunosuppressive microenvironment of HCC.