Nowadays, the safety issue has greatly hindered the development of large capacity lithium-ion batteries (LIBs), especially in electric vehicles applications. TiO2 is a kind of potential anode candidate for improving the safety of LIBs. However, it still needs to understand how to improve the performance of TiO2 anode in the practical applications. Herein, we design a contrast experiment by using three sizes of TiO2 hierarchical hollow microspheres (TiO2-HSs). The research results indicated that the cycling performance of TiO2-HSs anode can be affected by the size of microspheres, and the nanoparticles size of microspheres and crystallinity of TiO2 can affect their initial discharge capacity and lithium ion diffusion. And, the influence of microspheres size is more significant. This may provide a new strategy for improving the lithium-ion storage property of TiO2 anode material in the practical applications.