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Metal nanoparticles possess unique properties and usage patterns compared to traditional materials owing to their distinctive structures. In recent years, their application scenarios and dosages have considerably expanded. Biosynthetic nanoparticles, particularly those derived from endophytic fungi that help host organisms in adapting to heavy metal environments, hold substantial value and potential for application. This is largely attributed to their simplicity, cost-effectiveness, and energy efficiency. The present review provides an overview of the entire process of metal nanoparticle biosynthesis by plant endophytic fungi and illustrates various scenarios of their applications in oncology treatment. In addition to focusing on the preparation of metal nanoparticles using plant endophytic fungi, this review also explores the characterization of these nanoparticles and clarifies the synthesis mechanisms, including the synthetic pathways and the roles of fungal enzymes. It also comprehensively summarizes the application of biosynthetic metal nanoparticles in cancer, covering their role in diagnosis, enhancement of drug biocompatibility, and improvement of therapeutic efficacy. These nanoparticles exhibit toxicity toward cancer cells by generating reactive oxygen species and inducing oxidative stress, ultimately leading to the death of malignant cells. The biosynthesis of metal nanoparticles by plant endophytic fungi represents a promising, green, and environmentally friendly approach with potential applications in various fields, including cancer treatment, in the future.