Pipeline transmission is one of the most important ways of oil and gas transportation, and its safety is majorly threatened by fatigue fracture that generally occurs in heat-affected zone (HAZ) adjacent to the welded joint of the pipeline. Therefore, studying the fatigue properties of HAZ is important. In this work, the microstructure and mechanical properties in the HAZ of a X80 pipe were accurately simulated by thermal simulation. The fatigue life and crack growth rate of typical sub-HAZ were tested. Results showed that the fine grain HAZ had the lowest strength and fatigue life. By contrast, the coarse grain HAZ had the highest strength, but its fatigue life was lower than that of the intercritical HAZ. Furthermore, the microstructure of each sub-HAZ and its effect on fatigue properties were discussed in detail. The results will reveal the effects of microstructure and mechanical properties on the service safety of pipeline transportation.