In order to investigate the fatigue behavior of metals in nanoscale, a cyclic bending experiment is carried out using a nano-specimen. The specimen includes a copper film with a thickness of 20 nm constrained by highly rigid materials, which yields a high strain region with a size of a few nanometers near the interface edge. The specimen broke before the maximum load in the 7th cycle under fatigue (load range of 18 μN). The load-displacement curve shows nonlinear behavior and a distinct hysteresis loop, indicating plasticity in the Cu film. Reverse yielding appearing after the 2nd cycle suggests the development of a cyclic substructure in the Cu film. The cumulative plastic strain in the Cu film at fracture is more than three times larger than that under monotonic loading. These results indicate that the specimen breaks owing to fatigue of the Cu film on the nanoscale.