Rapid urbanization profoundly affects global biodiversity. How urbanization modifies soil biodiversity and perturbs nematodes remains limited. Here, we investigated soil nematodes in four land-use types: Parks, residential areas, natural forests, and maize fields across 12 cities in China. Urban parks and surrounding forests had similar nematode richness exceeding that in urban residential areas and surrounding farmlands. Nematode communities in parks and residential areas were, however, more homogenous than in forests and farmlands. Variations in nematode assemblages in both core urban and urban surroundings were mainly due to taxa replacement, indicating that nematodes were spatially isolated in cities. Urban residential areas were colonized by the lowest number of specialists (i.e., with narrow niche width) and smaller body sizes. Urban parks, conversely, served as hotspots for soil nematodes in cities. Together, our results indicate that urbanization processes reduce nematode diversity, with e.g., 30% loss in residential areas compared to forests, and homogenize soil nematode communities.