Abstract: Objective To observe the inhibitory ef fect of Yangyin Fuzheng Jiedu Decoction (YFJD) on a subcutaneous transplanted tumor model of hepatocellular carcinoma (HCC) driven by sgPten/c-Met, and to explore its possible molecular mechanism based on transcriptome sequencing. Methods The half maximal inhibitory concentration (IC50) of YFJD against the HepG2.2.15 and primary sgPten/c-Met cells was determined by the crystal violet assay. An orthotopic HCC model was established via hydrodynamic tail vein injection of the sgPten/c-Met/SB plasmid system in 5-week-old C57BL/6J mice. Four weeks after modeling, primary cells were isolated from the orthotopic tumor tissues and subcutaneously inoculated into the backs of mice to generate an ectopic subcutaneous tumor model. After tumor formation, YFJD was administered by gavage. Tumor volume and weight were measured. Subcutaneous tumor tissues were collected for transcriptome sequencing, and dif ferentially expressed genes were subjected to GO, KEGG, Reactome, and WikiPathways enrichment analyses. Results The IC50 of YFJD was 42.34 g/L for HepG2.2.15 cells and 55.66 g/L for sgPten/c-Met cells, indicating that HepG2.2.15 had a higher sensitivity to YFJD. At 21 days after modeling, compared with the control group, both the volume [(513.72 ± 229.86) mm3 vs. (937.35 ± 371.00) mm3; t = 3.21, P = 0.004] and weight [(0.62 ± 0.31) g vs. (1.22 ± 0.27) g; t = 4.64, P ＜ 0.001] of subcutaneous tumors in the YFJD group were signif icantly reduced. Transcriptome sequencing identif ied a total of 895 dif ferentially expressed genes, of which 590 were upregulated and 305 were downregulated. GO enrichment analysis indicated that the dif ferentially expressed genes were mainly involved in biological processes such as cell adhesion and extracellular matrix organization. KEGG enrichment analysis revealed that the Wnt signaling pathway was signif icantly enriched. Reactome analysis showed that the dif ferentially expressed genes were mainly enriched in immune-related pathways, including complement activation, antigen presentation, and programmed death-1 (PD-1) signaling. WikiPathways enrichment analysis demonstrated signif icant enrichment in terms such as the Wnt signaling pathway and Wnt signaling pathway and pluripotency. Conclusion YFJD could ef fectively inhibit the growth and proliferation of HCC cells and sgPten/c-Met-derived HCC subcutaneous transplanted tumors, and its mechanism may be related to negative regulation of the Wnt signaling pathway.