This study mainly describes the synthesis of Arginine-Glycine-Aspartic (RGD) peptide modification of copolymer: poly(lactic acid-co-L-lysine) (PLA-PLL- RGD) and preparation of mitoxantrone(DHAQ)-loaded PLA-PLL-RGD nanoparticles (DHAQ-NP), and researches the effects of exercise on antitumor efficacy of DHAQ-NP in hepatocellular carcinoma. Experimental results show that compared with DHAQ-only group, the DHAQ concentration of tumor in Exercise+DHAQ-NP and DHAQ-NP groups up-regulate significantly. Tumor volume and weight of mice were significantly decreased in the DHAQ-NP and Exercise+DHAQ-NP groups as compared to with the Control group. Suggesting that DHAQ-NP can effectively inhibit the growth of hepatocellular carcinoma. Compared with DHAQ-NP group, the DHAQ concentration, tumor volume of Exercise+DHAQ-NP group have no significant changes. This suggests exercise can not significantly influence the antitumor efficacy of DHAQ-NP. These studies are essential to fully understand the safety and application of exercise as a supportive intervention in targeted therapies.