Abstract: Alternative splicing (AS) is a critical mechanism in gene expression regulation, generating diverse mRNA isoforms and expanding protein diversity, which is essential for cellular physiological processes such as proliferation, metabolism, and apoptosis . In the field of oncology, AS has been significantly implicated in the regulation of various biological processes involved in tumorigenesis and cancer progression. Specific mutations or expression changes in splicing factors, such as SF3B1, SRSF2, and U2AF1, play key roles in multiple types of cancer. Targeting the spliceosome or splicing factors offers new strategies for cancer therapy, including the inhibition of spliceosome components and the regulation of alternative splicing patterns. This review discusses the mechanisms of RNA alternative splicing, its role in cancer, and the emerging strategies and prospects for its application in cancer treatment.