We reconstructed the late Holocene relative sea-level (RSL) evolution of the ancient harbour of Naples, one of the largest coastal conurbations in the Mediterranean. We carried out multiproxy investigations, coupling archaeological evidence with biological indicators. Our data robustly constrain 2000 yr of non-monotonic changes in sea level, chiefly controlled by the complex volcano-tectonic processes that characterize the area. Between ~200 BC and AD ~0, a subsidence rate of more than ~1.5 mm/yr enhanced the postglacial RSL rise, while negligible or moderate land uplift < ~0.5 mm/yr triggered a RSL stabilization during the Roman period (first five centuries AD). This stabilization was followed by a post-Roman enhancement of the sea-level rise when ground motion was negative, attested by a subsidence rate of ~0.5 to ~1 mm/yr. Our analysis seems to indicate very minor impacts of this nonmonotonic RSL evolution on the activities of the ancient harbour of Naples, which peaked from the third century BC to the second century AD. After this period, the progressive silting of the harbour basin made it impossible to safely navigate within the basin, leading to the progressive decline of the harbour.