Understanding the reciprocal link between changes in land use and transportation infrastructure is essential for sustainable development because of the consequences it has for both of these factors. The Czech Republic is a nation that possesses a robust railway system but is relatively understudied in this context. The purpose of this study is to give an in-depth analysis of the influence that railway networks have on changes to land use in the Czech Republic. The study focuses on critical railway lines linking Prague to Brno, Brno to Ostrava, and Prague to Plzeň using Geographic Information Systems (GIS) for spatial analysis and augmented by regression models. The investigation makes use of multi-temporal datasets, which incorporate topographical maps, land use classifications, and railway network data spanning three decades. According to the findings of the study, being in close proximity to a railway node acts as a driver for urban and industrial development while simultaneously causing a reduction in the amount of land used for agriculture. These findings give essential insights into the intricate interplay that exists between railway infrastructure and regional development, and they highlight the need to include land use planning into one's overall strategic planning.