DNA nanotechnology is a technique for generating programmable nucleic acid structures based on the Watson Crick base pairing principle. Nanostructures synthesized by DNA nanotechnology can not only interact with small molecules, nucleic acids, proteins, viruses and cancer cells, but also serve as nanocarriers to deliver different therapeutic drugs due to their high precision engineering design, unprecedented programmability and inherent biocompatibility. DNA origami, as an effective and multifunctional method to construct 2D and 3D programmable nanostructures, is a milestone in the development of DNA nanotechnology. Due to its highly controllable geometric shape space address-ability and easy chemical modification, DNA origami has great application potential in many fields. This review first introduces the basic principle of DNA origami, summarizes the development process of DNA origami according to the time sequence, introduces the software development process used to design DNA origami and compares its advantages and disadvantages. Drug loading of DNA nanostructures is described by category of drug loading, and drug release is described by release mode. Ideally, drug delivery carriers should be able to carry one or more drugs for collaborative treatment and penetrate the cell membrane and barrier to avoid harmful chemical and enzymatic degradation drugs, as well as adverse toxicity and immunogenicity, and show targeted and controlled drug release. Although drug delivery schemes based on DNA origami technology have not been applied in clinic, they are currently developing rapidly and show great potential in cancer treatment. This paper introduces the origin of DNA origami, starting with the basic principles and current progress, the methods of drug loading and releasing using DNA origami were summarized. Based on this technology, the future development trend, opportunities and challenges were prospected.