Abstract: The microbiome is a significant participant and driver of cancer development. Using traditional methods (such as antibiotics, probiotics, and microbiota transplantation) to regulate the microbiome has been proven to improve the therapeutic effect of cancer. But this type of method also has many limitations, such as the indirect damage to the symbiotic microbiota and the consistency of methodology. There is an urgent need to develop new technologies to solve these problems. Considering the success of nanotechnology in the field of cancer diagnosis and treatment, the use of nanotechnology to regulate the interaction between the microbiome and the tumor microenvironment is also expected to provide new and effective strategies for cancer treatment. In this review, we summarized the characteristics and advantages of various generations of nanotechnology, and reviewed the recent research related to the application of nanotechnology intervention to regulate the microbiome and its metabolites in cancer diagnosis and treatment, and discussed the challenges and future development prospects of this emerging field.