Objective To investigate the molecular mechanism by which licochalcone A inhibits colorectal cancer growth through regulation of the AMPK/mTOR signaling axis via long non-coding RNAs (LncRNAs).Methods Human colorectal cancer HCT116 cells and a nude mouse subcutaneous xenograft model were used. Cell proliferation was assessed by CCK-8 and EdU assays, and cell migration was evaluated by wound healing and Transwell assays. Quantitative PCR (qPCR) was employed to detect the expression levels of proliferation- and migration-related markers, and Western blotting was performed to analyze the expression of proteins related to the AMPK/mTOR pathway. Differential expressed LncRNAs in HCT116 cells after licochalcone A treatment were identified by LncRNA sequencing, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The in vivo anti-tumor effect of licochalcone A was validated using the nude mouse xenograft model. Multiple databases were integrated to identify potential targets of licochalcone A in colorectal cancer, and a protein-protein interaction (PPI) network was constructed to screen core targets, followed by GO functional annotation and KEGG pathway enrichment analysis. Furthermore, the function and mechanisms of LncRNA-CDC42BPA were verified by overexpression and RNA pull-down assays.Results Licochalcone A dose-dependently inhibited HCT116 cell proliferation, and suppressed DNA synthesis and cell migration. A total of 1 001 differentially expressed LncRNAs (556 downregulated and 445 upregulated) were identified after treatment, and GO/KEGG analyses showed enrichment in metabolic pathways and the AMPK/mTOR signaling pathway. Western blotting results confirmed that licochalcone A activated AMPK and inhibited mTOR. Network pharmacology identified 51 overlapping targets, and the PPI network indicated that AKT1, CASP3, and EGFR were core regulatory nodes. KEGG analysis further confirmed that the AMPK/mTOR pathway was a key pathway. Overexpression of LncRNA-CDC42BPA inhibited HCT116 cell proliferation and migration. Mechanistically, it regulated the AMPK/mTOR signaling pathway through binding to PTBP1 and FMR1.Conclusion Licochalcone A inhibits colorectal cancer cell proliferation, migration, and tumor growth in vivo by regulating the LncRNA-mediated AMPK/mTOR signaling axis.