As a natural barrier, the cell membrane must be crossed for drugs to enter cells and exert their therapeutic effects. As a potential drug delivery vehicle, carbon nanotubes (CNTs) have aroused widespread interest due to their unique physical and chemical properties. In this study, molecular dynamics simulation was employed to reveal the process of the model anticancer drug methotrexate (MTX) translocating through CNT nanochannels embedded in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membrane. From the perspective of translocation time-electric field intensity relationship, free energy calculation, density distribution of water and MTX during the translocation process, MTX conformational restrictions, etc., the translocation behavior and dynamics of MTX through membrane nanochannels were found to be co-regulated by the size of membrane nanochannels and the intensity of applied electric fields. These results may promote the design and application of biomimetic nanochannels and the associated drug delivery system in the biomedical field.