Tetrastigma hemsleyanum leaves (THL), a popular anticancer functional food in China, are used as nutrient or dietary supplement. However, the anticancer effects and the underlying mechanism of THL are still unknown. Thus, the phenolic profiles and anticancer activities of THL purified extract were investigated by a combination of chemical assays, LC-QTOF-MS/LC-QqQ-MS techniques, cell cultures and tumor-bearing mice model. It has been indicated that the TPC/TFC (421.63 and 343.38 mg/g DW) of THL extract were obviously increased after SPE purification, and the main phenolic components were apigenin and luteolin glycosides. Besides, THL could induce apoptosis by both of extrinsic death receptor pathway and intrinsic mitochondria pathway in HepG2 cells as well as inhibit the tumor growth, regulate Bcl-2 family proteins, and active the caspase family proteins in H22 tumor-bearing mice. Summarily, the findings clearly demonstrated that THL could be a potential functional food for the treatment and prevention of liver cancer.