The entrepreneurship cost is an important factor that affects the development of innovative entrepreneurship activities, but there are few quantitative studies on the cost of urban entrepreneurship. In this paper, the entrepreneurship cost is divided into six categories: cost of living, environmental cost, financing cost, operating cost, research and development cost and failure cost. Ten cities with active entrepreneurial activities in China were selected to carry out an empirical study on the entrepreneurship cost index for 2012-2016. The results show that the entrepreneurship cost in Shenzhen has been at a high level in 10 sample cities, that the cost of starting a business in Beijing and Shanghai has generally been on the rise in the past five years, and that the cost of starting a business in Changsha and Chengdu has been on the decline. The entrepreneurship cost in Chongqing has been kept low. The higher the level of economic development, the higher the explicit cost of entrepreneurship cost, but the relatively low hidden cost.