The booming applications of graphene in flexible electronics, mechanical structures, and biomedical sensors require robust mechanical and structural properties as a premise. With the ever-increasing demand for the long-term reliability of graphene-based devices and structures, the fatigue behavior of graphene necessitates careful investigation, especially its intrinsic and interfacial fatigue behavior. The fatigue concern of graphene is more stringent under extreme loading conditions in more complicated designs, such as at severe stress concentrations and abundant interfaces in flexible devices. Here we enabled the intrinsic fatigue study of suspended two-dimensional (2D) materials based on a modified atomic force microscopy technique. We discovered that monolayer and few-layer graphene also suffered fatigue, but they exhibited remarkable fatigue life of more than one billion cycles at large stress levels (e.g., at σ_mean=71 GPa and ∆σ=5.6 GPa), which is higher than any materials reported to date. Surprisingly, monolayer graphene did not reveal any obvious progressive damage during cyclic loading as manifested by its non-changing morphology and non-degraded mechanical properties. Molecular dynamics simulations further revealed bond reconfiguration near the defective site only immediately prior to fracture. Such non-progressive fatigue nature indicates the inapplicability of macroscopic fatigue mechanisms and challenges the fatigue definition at the atomic scale. A kinetic theory was proposed to explain the fatigue behavior of graphene and highlight the strong cyclic effect on reducing its lifetime. Graphene oxide, meanwhile, also exhibited ultrahigh fatigue resistance, but revealed clear progressive damage, similar to conventional fatigue mechanisms. Despite the record-high intrinsic fatigue life of graphene, we observed significant interfacial fatigue damage when introducing graphene-polymer contact. The significant elastic mismatch and weak van der Waals (vdW) interactions at the interface not only cause graphene buckling but also propagate the buckles under cyclic loading. The buckle propagation was revealed to follow an inverse Paris’ law. Moreover, cyclic loading through the vdW interfaces could also induce significant fracture of graphene even in tens of cycles, with the main fracture modes identified as in-plane shear and tear. These studies provide fundamental insights on the dynamic reliability of graphene and call for further fatigue studies of other 2D materials and their interfaces.