Anaplastic thyroid carcinoma (ATC) is a highly aggressive neoplasm with poor prognosis and limited therapeutic alternatives. Isoliquiritigenin (ISL), a bioactive isoflavonoid, has exhibited an antitumor activity across multiple tumor types
however, its precise anticancer mechanisms against ATC remain unexplored. In this study, the therapeutic effects of ISL on ATC cells and the potential mechanism were investigated by RNA-seq analysis and untargeted lipidomic analysis, combined with in vitro and in vivo experimental validation. The results showed that ISL effectively hindered the proliferation of ATC cells, inhibited cancer cell migration by up-regulating the level of E-cadherin and down-regulating the level of N-cadherin, and inhibited fatty acid synthesis by down-regulating the level of Sterol regulatory element binding transcription factor 1 (SREBF1) and its downstream lipid synthesis-related enzyme expression level. The underlying mechanism appears to involve a decrease in intracellular ATP levels induced by ISL and the activation of phosphorylated AMPK, thereby downregulating the expression of SREBF1, ultimately inhibiting cell proliferation, migration, and lipid synthesis. In vivo experiments further confirmed that ISL significantly retarded the growth of tumor xenografts in mice, diminished tumor cell proliferation, and reduced SREBF1 protein levels. This study suggests that ISL modulates lipogenesis and impedes cancer cell migration in ATC through the AMPK/SREBF1 signaling pathway.