The intrinsic high X-ray attenuation and insignificant biological toxicity of Bi-based nanomaterials make them a category of advanced materials in oncology. Bi-based two-dimensional nanomaterials have gained rapid development in cancer diagnosis and treatment owing to their adjustable bandgap structure, high specific surface area and strong NIR absorption. In addition to the single functional cancer diagnosis and treatment modalities, Bi-based two-dimensional nanomaterials have been certified for accomplishing multi-imaging guided multifunctional synergistic cancer therapies. In this review, we summarize the recent progress including controllable synthesis, defect engineering and surface modifications of Bi-based two-dimensional nanomaterials for cancer diagnosis and treatment in the past ten years. Their medical applications in cancer imaging and therapies are also presented. Finally, we discuss the potential challenges and future research priorities of Bi-based two-dimensional nanomaterials.