Metal-organic frameworks (MOFs), a class of porous crystalline materials formed by the self-assembly of metal ions/clusters and organic ligands, have shown broad application potential in the biomedical field due to their high specific surface area, precisely tunable pore structure, designable framework composition, and good biocompatibility. This paper traces the origin and development of MOFs, summarizes the contributions of the main promoters, and then systematically reviews the conventional synthesis and characterization methods of MOFs. Subsequently, it conducts an in-depth discussion around three application directions in the biomedical field: first, in the integration of cancer diagnosis and treatment, MOF-based treatment systems can integrate multiple modes such as chemotherapy, radiotherapy, photodynamic therapy, photothermal therapy, chemodynamic therapy, starvation therapy, and immunotherapy through single or combined strategies to exert a synergistic anti- tumor therapeutic effect; second, by constructing MOF-based carriers, including pH-responsive, GSH-responsive, and photo-responsive carriers, effective loading and precise delivery of drug molecules, including biological macromolecules, can be achieved; third, in the field of in vitro diagnosis, various MOF-based biomarker detection methods have been developed, providing technical means for the precise early diagnosis of diseases. Meanwhile, this paper deeply analyzes the key challenges that MOFs still face in clinical translation, including large-scale preparation, long-term stability, and biological safety assessment, and prospects the future development directions and application prospects.