Abstract: Select the panel data of 1470 companies recognized by the national high-tech enterprises from 2000 to 2017, establish a multi-regression analysis model to research the relationship between R&D investment, technology accumulation and innovation performance of high-tech enterprises, and the difference of the effect in the aspect that. whether registered in the high-tech zone or not and listed publicly or not. The results show that R&D investment has a significant positive impact on the innovation performance of high-tech enterprises. The internal R&D investment of the enterprise can produce better market performance than the financial R&D investment. The unique institutional advantages of high-tech enterprises registered in the high-tech zone can improve the efficiency of transformation from R&D investment to market performance, but the impact of financial funding is weaker than that outside the high-tech zone. High-tech enterprises with listed financing qualifications are less constrained by financing, making the positive impact of R&D investment on innovation performance more significant than unlisted. The conclusions of this paper can provide reference for the policy innovation of high-tech enterprises at this stage.