Abstract

Colorectal cancer (CRC) remains one of the most prevalent and deadly malignancies worldwide, requiring reliable biomarkers for detection. This study profiled tRNA-derived small RNAs in CRC tissues, identifying 1051 differentially expressed tsRNAs. Notably, i-tRF-Gly-GCC was significantly elevated in CRC tissues and patient sera compared to controls, with ROC analysis yielding an AUC > 0.7, underscoring its diagnostic potential. Weighted Gene Co-expression Network Analysis (WGCNA) revealed that i-tRF-Gly-GCC's target genes, including RAC2 , which was downregulated in the COAD and READ datasets, are closely related to CRC. Gene Ontology and KEGG pathway analyses further linked i-tRF-Gly-GCC to cancer-related processes. Functional experiments in HCT-116 cells demonstrated that i-tRF-Gly-GCC enhances cell proliferation and migration. These results highlight i-tRF-Gly-GCC as a novel non-invasive biomarker for CRC detection and a potential therapeutic target, advancing our understanding of tsRNA-mediated regulation in CRC and paving the way for clinical applications.