The variations of the microstructure and mechanical properties of medium-Mn steel after vanadium (V) microalloying with different contents were investigated. After a one-step intercritical annealing (IA) at 730 °C, the steel containing 0.04 wt.% of V exhibited excellent comprehensive properties. The steel maintained an ultimate tensile strength (UTS) of 1000 MPa while also exhibiting a total elongation (TEL) of 37% and a product of strength and plasticity (PSE) of 37.7 GPa%. V-microalloying improved the yield strength (YS) and UTS of the experimental steel by refining ferrite grains and precipitation strengthening, however, it deteriorated its plasticity, which is difficult to compensate for through grain refinement and due to the TRIP effect of retained austenite (RA). The largest amount of RA and the appropriate stability also make a significant contribution to the outstanding UTS of the steel containing 0.04 wt.% of V through the TRIP effect. However, the further increase of V content led to decreased RA content and stability, weakening the TRIP effect and resulting in a weaker strength ductility balance.