Objective To investigate the inhibitory effect of Agrimonia Pilosa Ledeb (APL) on transplanted tumor of liver cancer in nude mice and its mechanism. Methods HepG2 cells were subcutaneously injected into nude mice, which were randomly divided into 5 groups: Control group, Agrimonia Pilosa Ledeb low-dose group (APL 6g/kg), Agrimonia Pilosa Ledeb medium-dose group (APL 12g/kg), Agrimonia Pilosa Ledeb high-dose group (APL 24g/kg) and 5-fluorouracil group (5-FU 20mg/kg). The tumor weight was detected 30 days later. The changes of tumor volume at 30 days were detected. TUNEL was used to detect tumor apoptosis. The protein expression levels of Caspase-3, Bax, Bcl-2, LC3-I, LC3-II, Beclin1 and p62 were detected by Western blot. Ki67 and VEGF positive cells were detected by immunohistochemical staining. Results Compared with the Control group, the weight and volume of tumor were significantly decreased in APL dose groups and 5-Fu 20mg/kg group, and the apoptosis rate of tumor tissue was significantly increased, and the protein level of Bcl-2 was significantly decreased. LC3-II/LC3-I ratio and Beclin1 protein level were significantly increased, p62 protein level was significantly decreased, Ki67 and VEGF positive cell number were significantly decreased, the differences were statistically significant (P<0.05 or P<0.01). Conclusion APL may be involved in inducing apoptosis and autophagy of tumor cells and inhibiting proliferation of tumor cells by inhibiting the growth of transplanted tumor in nude mice.