Gly-tRF enhances LCSC-like properties and promotes HCC cells migration by targeting NDFIP2

Yongqiang Zhou,Zongli Fu,Lu Liu,Mengchao Yan,Xiaojing Song,Liming Hu,Jinjing Hu,Xun Li,Qiyu Zhang,Dan Zhu,Yue Chen,Yan Lin,Yongjian Wei

doi:10.1186/s12935-021-02102-8

Abstract

BackgroundAccumulating evidence demonstrates that tRFs (tRNA-derived small RNA fragments) and tiRNAs (tRNA-derived stress-induced RNA), an emerging category of regulatory RNA molecules derived from transfer RNAs (tRNAs), are dysregulated in in various human cancer types and play crucial roles. However, their roles and mechanisms in hepatocellular carcinoma (HCC) and liver cancer stem cells (LCSCs) are still unknown.MethodsThe expression of glycine tRNA-derived fragment (Gly-tRF) was measured by qRT-PCR. Flow cytometric analysis and sphere formation assays were used to determine the properties of LCSCs. Transwell assays and scratch wound assays were performed to detect HCC cell migration. Western blotting was conducted to evaluate the abundance change of Epithelial-mesenchymal transition (EMT)-related proteins. Dual luciferase reporter assays and signalling pathway analysis were performed to explore the underlying mechanism of Gly-tRF functions.ResultsGly-tRF was highly expressed in HCC cell lines and tumour tissues. Gly-tRF mimic increased the LCSC subpopulation proportion and LCSC-like cell properties. Gly-tRF mimic promoted HCC cell migration and EMT. Loss of Gly-tRF inhibited HCC cell migration and EMT. Mechanistically, Gly-tRF decreased the level of NDFIP2 mRNA by binding to the NDFIP2 mRNA 3′ UTR. Importantly, overexpression of NDFIP2 weakened the promotive effects of Gly-tRF on LCSC-like cell sphere formation and HCC cell migration. Signalling pathway analysis showed that Gly-tRF increased the abundance of phosphorylated AKT.ConclusionsGly-tRF enhances LCSC-like cell properties and promotes EMT by targeting NDFIP2 and activating the AKT signalling pathway. Gly-tRF plays tumor-promoting role in HCC and may lead to a potential therapeutic target for HCC.

Highlights

Accumulating evidence demonstrates that TRNA-derived small RNA fragments (tRFs) and tiRNAs, an emerging category of regulatory RNA molecules derived from transfer RNAs, are dysregulated in in various human cancer types and play crucial roles
A recent study showed that glycine transfer RNA (tRNA)-derived fragment (Gly-tRF) expression is upregulated in ethanol-fed mice and promotes alcoholic fatty liver disease (AFLD) [21]
We observed that overexpression of Nedd4 family interacting protein 2 (NDFIP2) weakened the promotive effects of Gly-tRF on Epithelial-mesenchymal transition (EMT) and liver cancer stem cells (LCSCs)-like cell sphere formation ability

Summary

Introduction

Accumulating evidence demonstrates that tRFs (tRNA-derived small RNA fragments) and tiRNAs (tRNA-derived stress-induced RNA), an emerging category of regulatory RNA molecules derived from transfer RNAs (tRNAs), are dysregulated in in various human cancer types and play crucial roles. Their roles and mechanisms in hepatocellular carcinoma (HCC) and liver cancer stem cells (LCSCs) are still unknown. Accumulating evidence shows that tRFs and tiRNAs play crucial roles in human cancers, including breast cancer [12,13,14,15], prostate cancer [16, 17], and colorectal cancer [18, 19], by participating in multiple biological functions, including gene expression and silencing, translation regulation and epigenetic regulation [20].

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