Journal Article

LncRNA FTO-IT1 promotes glycolysis and progression of hepatocellular carcinoma through modulating FTO-mediated N6-methyladenosine modification on GLUT1 and PKM2

Hu, Yuhang, Huang, Yan, Zhuo, Wenfeng, Wang, Jie, Wang, Hongda, Hu, Ping, Han, Shengbo, Xiong, Hewei, Zhao, Yong, Wang, Decai, Wang, Fan, Zeng, Zhu, Zhao, Gang, Lv, Guozheng

2023

Overview

Background Long non-coding RNAs (LncRNAs) have been extensively studied to play essential roles in tumor progression. However, more in-depth studies are waiting to be solved on how lncRNAs regulate the progression of hepatocellular carcinoma (HCC). Methods Different expression levels of lncRNAs in HCC cells were compared by analysis of Gene Expression Omnibus and The Cancer Genome Atlas databases. The effects of lncRNA FTO Intronic Transcript 1 (FTO-IT1) on HCC cells were assessed by gain- and loss-of-function experiments. Colony formation assay, Edu assay, glucose uptake and lactic acid production assay were performed to evaluate the regulation of proliferation and glycolysis of HCC cells by FTO-IT1. The binding between protein interleukin enhancer binding factor 2/3 (ILF2/ILF3) and FTO-IT1 was determined by RNA pull-down, mass spectroscopy and RNA immunoprecipitation experiments. RNA stability assay, quantitative reverse transcription PCR and Western blot were employed to determine the regulatory mechanisms of FTO-IT1 on fat mass and obesity-associated (FTO). Methylated RNA immunoprecipitation assay was used to assessed the regulation of key enzymes of glycolysis by FTO. The role of FTO-IT1/FTO in vivo was confirmed via xenograft tumor model. Results LncRNA FTO-IT1, an intronic region transcript of FTO gene, was highly expressed in HCC and associated with poor prognosis of patients with HCC. FTO-IT1 was related to proliferation and glycolysis of HCC cells, and contributed to the malignant progression of HCC by promoting glycolysis. Mechanistically, FTO-IT1 induced stabilization of FTO mRNA by recruiting ILF2/ILF3 protein complex to 3’UTR of FTO mRNA. As a demethylase for N 6 -methyladenosine (m 6 A), FTO decreased m 6 A modification on mRNAs of glycolysis associated genes including GLUT1, PKM2, and c-Myc which alleviated the YTH N6-methyladenosine RNA binding protein 2 (YTHDF2)-mediated mRNA degradation. Therefore, the upregulated expression of FTO-IT1 leaded to overexpression of GLUT1, PKM2, and c-Myc by which enhanced glycolysis of HCC. Meanwhile, it was found that c-Myc transcriptional regulated expression of FTO-IT1 by binding to its promoter area under hypo-glucose condition, forming a reciprocal loop between c-Myc and FTO-IT1. Conclusions This study identified an important role of the FTO-IT1/FTO axis mediated m 6 A modification of glycolytic genes contributed to glycolysis and tumorigenesis of HCC, and FTO-IT1 might be served as a new therapeutic target for HCC.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Alpha-Ketoglutarate-Dependent Dioxygenase FTO - genetics / Alpha-Ketoglutarate-Dependent Dioxygenase FTO - metabolism / Animals / Antibodies / Apoptosis / Biomedical and Life Sciences / Biomedicine / Cancer / Cancer Research / Carcinoma, Hepatocellular - pathology / Cell cycle / Cell Line, Tumor / Cell Proliferation - genetics / Comparative analysis / Development and progression / Dextrose / Disease Models, Animal / Enzymes / Ethanol / Ethylenediaminetetraacetic acid / FTO / FTO-IT1 / Gene Expression Regulation, Neoplastic / Genes / Genetic aspects / Genetic transcription / Genomes / Genomics / Glucose / Glucose metabolism / Glucose Transporter Type 1 - genetics / Glycolysis / Hepatocellular carcinoma / Hepatoma / Humans / Immunology / Laboratory animals / Lactic acid / Liver cancer / Liver Neoplasms - pathology / m6A / Mass spectrometry / Medical prognosis / Messenger RNA / Metastasis / Oncology / Plasmids / Protein binding / Proteins / RNA, Long Noncoding - genetics / RNA, Long Noncoding - metabolism / RNA, Messenger - metabolism / Scientific imaging / Thyroid Hormone-Binding Proteins