DNA molecules have shown unique advantages and high potential in enhancing drug targeting and reducing drug toxicity because of their unique biocompatibility and programmability. And spurred by the development of tumor microenvironment research and the development of environmentally responsive DNA triggers, many DNA nanostructure delivery systems based on tumor microenvironment have been reported in recent years. These DNA nanostructures combine the favorable biodistribution and pharmacokinetic properties of nanodelivery vehicles and the rapid diffusion and penetration properties of smaller drug cargos. By targeting a wide range of tumor habitats rather than tumor specific receptors, the strategy is likely to overcome the problem of tumor heterogeneity and can be used to design nanoparticles for diagnosis and treatment of multiple solid tumors. They can be transported steadily in the blood circulation, releasing drugs under the unique microenvironmental stimulation of tumor tissue, effectively controlling the drug release site and release speed, and greatly reducing the toxic and side effects of tumor treatment. In this article, the latest research progress of tumor microenvironment responsive DNA nanostructure drug delivery systems is reviewed from five aspects of pH responsive systems, GSH responsive systems, ATP responsive systems, enzyme responsive systems, and antigen responsive systems. The design strategy and responsive release mechanism of corresponding DNA nanocarriers are discussed in great detail. In addition, the prospects and main challenges in this area are also discussed.