Abstract: A long-span railway cable-stayed bridge was regarded as an engineering example,then the effect of the stiffness of girder,cables and auxiliary piers was analyzed by changing structural stiffness. It is shown by analysis results that the increase of truss width can significantly increase the lateral bending stiffness of the bridge, but have a limited impact on train running performance; Increasing truss height or the cable cross section area can both notably increase the vertical bending fundamental frequency and reduce the response of vehicle-bridge systems; Local bridge deck system contributes little to the overall bridge stiffness and has a limited impact on the vehicle response; Auxiliary pier can improve the vertical stiffness of the bridge, reduce the vehicle vertical acceleration and the vertical rotation angle at girder ends.