The invention discloses an integrated microfluidic chip and a three-dimensional tumor location, construction and recycling method. The disclosed chip is composed of a sample flow layer and a pneumatic control layer; the sample flow layer is provided with an inlet, micro-cavities, outlets and a micro pipeline network, and the pneumatic control layer is provided with a gas inlet, pneumatic control structures and an end-closed micro pipeline network. The pneumatic control structures are connected with the gas inlet through the end-closed micro pipeline network. The height of the pneumatic control structures is greater than that of the end-closed micro pipeline network. Tumor cell accurate locating capture, three-dimensional tumor forming and online analysis and recycling operations can be continuously completed in the single microfluidic chip. The operations have extremely strong time and space control and flexibility, and are convenient for multidisciplinary research scholars to master; and the chip has the characteristics of simple and fast operations, low sample consumption, accurate cell location, high efficiency, high throughput and the like, and is suitable for construction and analysis operations on various three-dimensional tumors.