The aim of the article is to contribute to the discussion of effectiveness of construction of high-speed lines (HSR) in the Czech Republic with an emphasis on the perception of their impacts on the spatial mobility of labor force. The concrete approach to impacts assessment is based on the potential availability of twelve urban centers on planned lines and their attractiveness for labor mobility, using data on commuting to the 2011 census. The results document strategic position of Prague as the main commuting center, which relevant indicators significantly exceed all other labor mobility centres (the second most important center Brno is about half the importance). This fact was confirmed not only by the analysis of gradient labor areas, but also by evaluating of commuting relations among relevant centers. The system assessment of potential impacts of HSR on labor markets is then carried out using the model of marginal rate of labor mobility, where it is possible to count on positive impacts except for Prague on two other commuter centers, Brno and Pilsen. In the case of other analyzed centers, then we can count on increasing the potential of a trip for work to the above-mentioned centers.