The Western Liaohe River Basin, rich in prehistoric cultures, is one of the significant origins of Chinese civilization. However, what is the subsistence basis that supports the development of prehistoric cultures in this region? More importantly, which role did agriculture play in the development of various prehistoric cultures? Which factors drive the diachronic evolution in subsistence strategies? These issues are worthy of study. Based on this, we systematically collected zooarchaeological, archaeobotanical, and stable isotope data from various prehistoric archeological cultural sites spanning from the Neolithic Age to the Bronze Age in the Western Liaohe River Basin, attempting to explore their economic characteristics, evolution process as well as driving factor. The study found that in the Western Liaohe River Basin, the cultivation of millet crops may have begun as early as the Xiaohexi cultural period, and domestic pigs and dogs may have occurred in the Xinglongwa cultural period (Xionglongwa and Xionglonggou site respectively). However, during Hongshan cultural period, collection and hunting (cervids) remained the main subsistence strategy. In the Lower Xiajiadian cultural period, millet agriculture and domestic pig breeding replaced gathering and hunting, as the predominant subsistence strategy. This subsistence pattern was also adopted by subsequent Upper Xiajiadian cultural populations, and in some areas, animal husbandry developed during this period. We analyze that the evolution of the subsistence strategies of the prehistoric humans in this region may be related with Holocene climate change. Meanwhile, other factors such as improvements in production technology, population pressure and the environmental impacts they brought may also play a role. This study reveals the evolution process of subsistence strategies of prehistoric cultures in the Western Liaohe River Basin, and is of great significance in understanding the relationship between subsistence evolution of ancient populations and environmental changes in northern China.