Actinomycin D inhibits liver cancer invasion and metastasis by suppressing the Wnt/β-catenin signaling pathway and epithelial-mesenchymal transition

<x>Abstract</x> <sec> <title>Objective:</title> To investigate actinomycin D’s effects on liver cancer progression, explore its underlying mechanisms, focusing on epithelial-mesenchymal transition (EMT) and Wnt/β-catenin signaling, offer new insights and references for identifying low toxicity traditional Chinese medicine (TCM) monomers with the same targets but lower toxicity or developing compound formulations to enhance pathway blocking effects, and to innovate and develop integrated Chinese and western medicine treatment strategies. </sec> <sec> <title>Methods:</title> Human liver cancer cells (HepG2) were treated with actinomycin D for <italic>in vitro</italic> assays. Cell proliferation was assessed <italic>via</italic> CCK-8, while transwell assays evaluated invasion and migration. Western blot analyzed expression of EMT markers (E-cadherin, β-catenin, Twist1). A nude mouse metastatic model using HepG2 cells was established for <italic>in vivo</italic> validation, with metastasis efficiency and molecular markers compared between treatment and control groups. </sec> <sec> <title>Results:</title> Actinomycin D suppressed liver cancer cell proliferation in a dose-dependent and time-dependent manner (CCK-8 assay) and inhibited cell invasion and migration (assessed by transwell assay). Western blot analysis showed that actinomycin D downregulated β-catenin and Twist1 while upregulating E-cadherin, indicating suppression of EMT. <italic>In vivo</italic> experiments demonstrated that actinomycin D significantly reduced metastatic efficiency in mice, consistent with decreased β-catenin/Twist1 expression and increased E-cadherin levels in tumor tissues. </sec> <sec> <title>Conclusions:</title> Actinomycin D suppresses liver cancer invasion and metastasis by inhibiting the Wnt/β-catenin pathway and EMT progression. These findings highlight its dual antitumor role in both proliferation inhibition and metastasis suppression, providing mechanistic insights for its clinical applications. </sec>