Abstract
BACKGROUND: Transfer RNA-derived small RNAs (tsRNAs) are a recently discovered class of non-coding RNAs with aberrant expression in various cancers. Substantial evidence implicates tsRNAs in the initiation and progression of colorectal cancer (CRC). This study aimed to investigate the diagnostic and prognostic potential of a specific tsRNA, tRF5-23-GlyTCC-2, in CRC. METHODS: We identified tRF5-23-GlyTCC-2 via high-throughput RNA sequencing and validated its expression using qRT-PCR. Associations between tRF5-23-GlyTCC-2 expression, clinicopathological features, and patient survival were assessed with Chi-square and Kaplan-Meier analyses. Its diagnostic performance was evaluated by ROC curve analysis. Functional roles in CRC were examined using colony formation assays and xenograft mouse models. RESULTS: Expression of tRF5-23-GlyTCC-2 was significantly downregulated in CRC tissues (P = 0.0009) and serum (P < 0.0001) compared to controls. It effectively discriminated CRC patients from healthy individuals and those with colorectal polyps, and served as a strong predictor of poor prognosis. Low tRF5-23-GlyTCC-2 levels were correlated with advanced invasion, metastasis (P = 0.0153), and poor prognosis (P = 0.004). ROC analysis demonstrated its superior diagnostic accuracy over traditional biomarkers (AUC = 0.8628), and its combination with CEA further improved the diagnostic performance (AUC = 0.9077). Both in vitro colony formation assays and in vivo xenograft models confirmed its tumor-suppressive function by inhibiting tumor growth and progression. CONCLUSION: Serum tRF5-23-GlyTCC-2 exhibits high diagnostic accuracy, and its combination with CEA achieves superior sensitivity (84%), highlighting its potential as a powerful non-invasive biomarker to improve CRC detection and prognosis prediction.