In order to reveal the dislocation evolution law of body-centered cubic axle steel EA4T during cyclic deformation and provide an experimental basis for the subsequent construction of cyclic constitutive models based on microscopic physical mechanisms, macroscopic deformation experiments were first conducted on axle steel EA4T, including monotonic tensile experiments under different deformation amounts, symmetric strain cycling experiments under a different number of cycles, and ratcheting deformation experiments under a different number of cycles. Then, systematic observations of different samples at different deformation stages were conducted using a Transmission Electron Microscope (TEM) to investigate the dislocation configuration and evolution during strain cycling and ratcheting deformation. The observed results show that the dislocation evolution law of axle steel EA4T during the uniaxial tensile experiment, symmetrical strain cycling, and ratcheting deformation is basically the same, and the dislocation density increases with the increase in plastic deformation and number of cycles. The dislocation configuration gradually develops from low-density dislocation configurations such as dislocation lines and dislocation pileups to high-density dislocation configurations such as severe dislocation tangles and dislocation walls. The microscopic mechanism of the uniaxial ratcheting evolution of axle steel EA4T can be qualitatively explained by the dislocation configuration and evolution.