The effect of varying microstructural parameters on the cyclic behaviour of dual-phase steels was studied on the basis of experimental and micromechanical finite-element simulated results. The initial bainitic morphology of as-received 20MnMoNi55 steel was transformed into ferrite and martensite through proper inter-critical heat treatment procedures. Strain-controlled low cycle fatigue tests were conducted at room temperature with different strain amplitudes at a specific strain rate of 10−3/s. The cyclic stress–strain curve, obtained through joining the peak stresses of hysteresis loops corresponding to different strain amplitude, shows an increase in strain hardening with an increase in volume fraction of martensite. Whereas the rate of cyclic softening, considering the decrease in stress amplitude with respect to elapsed cycles, increases with increasing strain amplitude. Inclusive of all affecting microstructural parameters, an original microstructure-based representative volume element associated with a crystal plasticity-based material model was adopted for conducting micromechanical finite-element simulation. In addition to several parameters associated with a crystal plasticity model, consideration of initial geometrically necessary dislocation density in constituent phases resulted in the accurate prediction of a hysteresis loop at low strain amplitude as compared with the experimental results. A variation of stress triaxiality built up in ferrite matrix with martensite fraction along with deformation inhomogeneity between ferrite and martensite was also observed through a strain partitioning phenomenon obtained from finite-element simulated results.