Abstract: The directional migration of cells guided by bioelectrical signals plays an important role in the major physiological and pathological processes of multicellular organisms. However, the mechanism of this is not clear. Using in vitro direct current stimulation to explore the mechanism of cell directional migration is a common method to study the bioelectrical signal to guide cell behavior. The effect of mTOR signaling pathway on the electrotaxis of human promyelocytic leukemia cell line HL60 was studied in this paper. The results showed that in a DC electric field of 300 mV/mm, HL60 cells showed obvious electrotaxis towards the cathode of the electric field, and the electrotaxis index and trajectory velocity were 0.89 ± 0.04 and 9.72 ± 0.38 μm/min, respectively. There was no restrictive difference between the electrotaxis of Rictor knockdown mutant and wild type. However, after HL60 cells were treated with high concentration of mTOR specific inhibitor PP242, the electrotaxis index of cells in DC electric field decreased to 0.82 ± 0.06, and the movement speed was also significantly inhibited. These results suggest that mTOR signaling pathway plays a role in the electrotaxis of HL60 cells, but Rictor is not a key gene. This result provides a theoretical basis for the development of bioelectric field-dependent targeted drugs.