Based on the case study of Guiyang City, the minimum cumulative resistance model integrating construction land source, ecological rigid constraints and ecological function type resistance factor, was built by use of costdistance analysis of urban spatial expansion resistance value through ArcGIS 9.3 software in this paper. Then, the ecological resistance of city spatial expansion of Guiyang from 2010 was simulated dynamically and the ecological suitability classification of city spatial expansion was assessed. According to the conflict between the newly increased city construction land in 2014 and its ecological suitability, the unreasonable city land spatial allocation was discussed also. The results showed that the ecological suitability zonation and the city expansion in the study area were basically consistent during 2010-2014, but the conflict between the new city construction and its land ecological suitability was more serious. The ecological conflict area accounted for 58.2% of the new city construction sites, 35.4% of which happened in the ecological control area, 13.9% in the limited development area and 8.9% in the prohibition development area. The intensification of ecological land use conflict would impair the ecological service function and ecological safety, so this paper put forward the city spatial expansion optimal path to preserve the ecological land and improve the construction land space pattern of Guiyang City so as to ensure its ecological safety.