tsRNAs sequencing of FFPE tissues reveals i-tRF-Gly-GCC as a biomarker for colorectal cancer

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. • We identified tRNA-Gly (GCC)-derived tsRNA as up-regulated markers in colorectal cancer samples using tsRNA sequencing. • tRNA-Gly (GCC)-derived i-tRF-Gly-GCC in CRC was high in colorectal cancer tissue and serum compared to the control. • WGCNA analysis showed that i-tRF-Gly-GCC target genes were enriched in cancer-associated pathways. • The mechanism was experimentally validated, demonstrating that i-tRF-Gly-GCC specifically binds to the RAC2 gene. • Function validation experiments confirmed that i-tRF-Gly-GCC significantly enhanced cell proliferation and migration.