5’-transfer RNA halve-lysine-CTT as a promising biomarker for early detection of hepatocellular carcinoma

Abstract

BACKGROUND

Hepatocellular carcinoma (HCC) is a globally prevalent malignancy associated with high morbidity and mortality. Transfer RNA (tRNA)-derived small RNAs (tsRNAs), a class of small non-coding RNAs originating from tRNA, have emerged as potential therapeutic targets in cancers, including HCC. However, the specific tsRNAs involved in HCC and their precise mechanisms remain largely unknown. In this study, we identify and characterize specific tsRNAs involved in the development and progression of HCC, discovering their potential as novel biomarkers for early detection and potential therapeutic targets.

AIM

To investigate differentially expressed tsRNAs in HCC, identify potential biomarkers, and elucidate the functions and mechanisms of tsRNAs in HCC.

METHODS

Differentially expressed tsRNAs in Barcelona Clinic Liver Cancer 0/A-stage HCC tissues were identified through high-throughput sequencing. Agarose gel electrophoresis, Sanger sequencing, and quantitative polymerase chain reaction were conducted to detect 5’-tRNA halve (tiRNA)-lysine (Lys)-CTT in tissues and serum samples. The diagnostic performance of 5’-tiRNA-Lys-CTT was evaluated using receiver operating characteristic analysis. HCC cell proliferation was examined using the Cell Counting Kit-8 assay, colony formation assay, and 5-ethynyl-2’-deoxyuridine staining. Additionally, the migratory capability of HCC cells was investigated using Transwell assays.

RESULTS

The 5’-tiRNA-Lys-CTT demonstrated excellent stability and can be easily detected. Its expression was significantly upregulated in 50 HCC tissues, 110 HCC serum samples, and 5 HCC cell lines vs control groups, and the differences were all significant. This elevated expression was strongly associated with clinicopathological characteristics, particularly tumor size, Barcelona Clinic Liver Cancer stage, and cirrhosis of the liver. Receiver operating characteristic analysis revealed superior detection efficiency of 5’-tiRNA-Lys-CTT exhibits for early-stage HCC compared to established markers. Functional assays revealed that 5’-tiRNA-Lys-CTT overexpression promoted cell proliferation and migration, while its inhibition had the opposite effect. Bioinformatics predictions suggest that 5’-tiRNA-Lys-CTT may influence the development and progression of liver cancer by targeting downstream mRNA via metabolic pathways, cancer pathways, and HCC-specific pathways.

CONCLUSION

The 5’-tiRNA-Lys-CTT levels were higher in early HCC patients. 5’-tiRNA-Lys-CTT is a promising diagnostic biomarker for early-stage HCC and may play an oncogenic role in HCC by interacting with downstream mRNA targets via specific pathways.

Keywords: 5’-transfer RNA halve-lysine-CTT; Hepatocellular carcinoma; Transfer RNA-derived small RNAs; Transfer RNA-derived fragments; Diagnosis biomarker

Core Tip: Through high-throughput sequencing, we found transfer RNA (tRNA)-derived small RNA differentially expressed in early-stage hepatocellular carcinoma (HCC) tissues. 5’-tRNA halve (tiRNA)-lysine (Lys)-CTT, one of the tRNA-derived small RNAs, had great diagnostic efficacy and potential as new “liquid biopsy” biomarkers for the diagnosis of early stage of HCC. We speculated that 5’-tiRNA-Lys-CTT may be involved in multiple cancer-related signaling pathways, and would like to investigate the specific mechanisms of 5’-tiRNA-Lys-CTT in HCC in future studies.