Transfer RNA (tRNA)-derived small non-coding RNAs (tsRNAs) are generated through specific endonucleolytic cleavage of mature or precursor tRNA at different sites and exist widely in prokaryotic and eukaryotic transcriptome, and are a new class of gene expression regulators with a variety of biological functions. tsRNAs participate in some physiological and pathological processes, including stress response, protein translation, ribosomal biosynthesis and regulating immunity. Dysregulation of tsRNAs is closely related with the occurrence and development of many human diseases. This article reviewed the advance in study on tsRNAs in gastrointestinal tumors.