Hepatocellular carcinoma (HCC) is a malignant tumor of the digestive system, influenced by various factors. Due to its subtle clinical symptoms, it delayed results in poor prognosis and limited treatment options. Cellular senescence, characterized by stable growth arrest, is closely linked to tumor proliferation inhibition, making it a promising therapeutic strategy for HCC. However, the role of Dual Specificity Phosphatase 3 (DUSP3) in HCC-induced senescence and its underlying mechanisms remain poorly understood. Our preliminary data show a marked upregulation of DUSP3 in HCC tissues compared to adjacent group. Additionally, DUSP3 knockdown induced senescence in HCC cells in vitro. Further investigation revealed that inhibiting Notch1 reversed the senescence induced by DUSP3 knockdown in these cells. Thus, targeting DUSP3 to activate the Notch1 pathway and induction of senescence as a promising anti-tumor strategy.