Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma

Abstract Background RNA 5‐methylcytosine (m5C) plays an important role in the progression of hepatocellular carcinoma (HCC). Dysregulation of ferroptosis is closely associated with HCC. However, the effect of the epigenetic mRNA m5C modification on ferroptosis in HCC remains unclear. Methods In this...

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Main Authors: Ouwen Li, Ke An, Han Wang, Xianbin Li, Yueqin Wang, Lan Huang, Yue Du, Nuo Qin, Jiasheng Dong, Jingyao Wei, Ranran Sun, Yong Shi, Yanjia Guo, Xiangyi Sun, Ying Yang, Yun‐Gui Yang, Quancheng Kan, Xin Tian
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Language:English
Published: Wiley 2025-03-01
Series:Clinical and Translational Medicine
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Online Access:https://doi.org/10.1002/ctm2.70270
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author Ouwen Li
Ke An
Han Wang
Xianbin Li
Yueqin Wang
Lan Huang
Yue Du
Nuo Qin
Jiasheng Dong
Jingyao Wei
Ranran Sun
Yong Shi
Yanjia Guo
Xiangyi Sun
Ying Yang
Yun‐Gui Yang
Quancheng Kan
Xin Tian
author_facet Ouwen Li
Ke An
Han Wang
Xianbin Li
Yueqin Wang
Lan Huang
Yue Du
Nuo Qin
Jiasheng Dong
Jingyao Wei
Ranran Sun
Yong Shi
Yanjia Guo
Xiangyi Sun
Ying Yang
Yun‐Gui Yang
Quancheng Kan
Xin Tian
author_sort Ouwen Li
collection DOAJ
description Abstract Background RNA 5‐methylcytosine (m5C) plays an important role in the progression of hepatocellular carcinoma (HCC). Dysregulation of ferroptosis is closely associated with HCC. However, the effect of the epigenetic mRNA m5C modification on ferroptosis in HCC remains unclear. Methods In this study, ferroptosis was evaluated by detecting lipid reactive oxygen species (lipid ROS), ferrous ion and 4‐hydroxynonenal (4‐HNE) in xenograft mouse model, diethylnitrosamine (DEN)‐initiated HCC model and so forth. The regulatory mechanisms of YBX1 in mRNA translation were elucidated using RNA sequencing, ribosome sequencing, RNA immunoprecipitation (RIP)‐sequencing, bisulphite sequencing and immunoprecipitation (IP)–mass spectrometry assays. Dual‐luciferase reporter, RIP‐qPCR, Co‐IP, RNA pulldown and methylated RNA immunoprecipitation (MeRIP)‐quantitative polymerase chain reaction (qPCR) assays were performed to validate the mechanism of YBX1 in regulating mRNA translation by m5C modification. Results Here, we found that YBX1 promoted the translation of Ring Finger Protein 115 (RNF115) mRNA through m5C modification, thereby inhibiting ferroptosis and promoting HCC development. Moreover, RNF115 was identified as an E3 ubiquitin ligase for dihydroorotate dehydrogenase (DHODH), promoting Lys27 (K27) ubiquitination and inhibiting its autophagic degradation to counteract ferroptosis. In addition, YBX1 bound to the m5C modification sites of RNF115 3′‐untranslated region (UTR) and interacted with Eukaryotic Translation Initiation Factor 4A1 (EIF4A1) to bridge the 5′‐UTR regions, promoting mRNA circularisation and translation, while NOP2/Sun RNA methyltransferase 2 (NSUN2) was identified as responsible for m5C modification of RNF115 mRNA in HCC. Conclusions The current work revealed that YBX1 promoted RNF115 mRNA translation in an m5C‐dependent manner, thereby regulating DHODH ubiquitination and expression to suppress ferroptosis. This research sheds light on the mechanism of YBX1 in m5C‐modified mRNAs translation and ferroptosis, highlighting its promise as a biomarker for prognosis and a target for therapy in HCC. Key points YBX1 inhibits ferroptosis in HCC by regulating the RNF115‐DHODH axis. RNF115, an E3 ligase, mediates K27 ubiquitination and autophagic degradation of DHODH. YBX1 binds to the m5C sites of RNF115 mRNA 3′‐UTR and interacts with EIF4A1 to bridge the 5′‐UTR, promoting mRNA circularisation and translation. High expression of YBX1/RNF115 predicts the poor overall survival in HCC.
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spelling doaj-art-24a08e7c80434e6695033f8dbad2daf52025-08-20T03:07:28ZengWileyClinical and Translational Medicine2001-13262025-03-01153n/an/a10.1002/ctm2.70270Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinomaOuwen Li0Ke An1Han Wang2Xianbin Li3Yueqin Wang4Lan Huang5Yue Du6Nuo Qin7Jiasheng Dong8Jingyao Wei9Ranran Sun10Yong Shi11Yanjia Guo12Xiangyi Sun13Ying Yang14Yun‐Gui Yang15Quancheng Kan16Xin Tian17Department of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaTranslational Medicine Center The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Infectious Diseases The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaKey Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, Beijing Institute of Genomics, Chinese Academy of Sciences China National Center for Bioinformation Beijing ChinaKey Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, Beijing Institute of Genomics, Chinese Academy of Sciences China National Center for Bioinformation Beijing ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaDepartment of Pharmacy The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan ChinaAbstract Background RNA 5‐methylcytosine (m5C) plays an important role in the progression of hepatocellular carcinoma (HCC). Dysregulation of ferroptosis is closely associated with HCC. However, the effect of the epigenetic mRNA m5C modification on ferroptosis in HCC remains unclear. Methods In this study, ferroptosis was evaluated by detecting lipid reactive oxygen species (lipid ROS), ferrous ion and 4‐hydroxynonenal (4‐HNE) in xenograft mouse model, diethylnitrosamine (DEN)‐initiated HCC model and so forth. The regulatory mechanisms of YBX1 in mRNA translation were elucidated using RNA sequencing, ribosome sequencing, RNA immunoprecipitation (RIP)‐sequencing, bisulphite sequencing and immunoprecipitation (IP)–mass spectrometry assays. Dual‐luciferase reporter, RIP‐qPCR, Co‐IP, RNA pulldown and methylated RNA immunoprecipitation (MeRIP)‐quantitative polymerase chain reaction (qPCR) assays were performed to validate the mechanism of YBX1 in regulating mRNA translation by m5C modification. Results Here, we found that YBX1 promoted the translation of Ring Finger Protein 115 (RNF115) mRNA through m5C modification, thereby inhibiting ferroptosis and promoting HCC development. Moreover, RNF115 was identified as an E3 ubiquitin ligase for dihydroorotate dehydrogenase (DHODH), promoting Lys27 (K27) ubiquitination and inhibiting its autophagic degradation to counteract ferroptosis. In addition, YBX1 bound to the m5C modification sites of RNF115 3′‐untranslated region (UTR) and interacted with Eukaryotic Translation Initiation Factor 4A1 (EIF4A1) to bridge the 5′‐UTR regions, promoting mRNA circularisation and translation, while NOP2/Sun RNA methyltransferase 2 (NSUN2) was identified as responsible for m5C modification of RNF115 mRNA in HCC. Conclusions The current work revealed that YBX1 promoted RNF115 mRNA translation in an m5C‐dependent manner, thereby regulating DHODH ubiquitination and expression to suppress ferroptosis. This research sheds light on the mechanism of YBX1 in m5C‐modified mRNAs translation and ferroptosis, highlighting its promise as a biomarker for prognosis and a target for therapy in HCC. Key points YBX1 inhibits ferroptosis in HCC by regulating the RNF115‐DHODH axis. RNF115, an E3 ligase, mediates K27 ubiquitination and autophagic degradation of DHODH. YBX1 binds to the m5C sites of RNF115 mRNA 3′‐UTR and interacts with EIF4A1 to bridge the 5′‐UTR, promoting mRNA circularisation and translation. High expression of YBX1/RNF115 predicts the poor overall survival in HCC.https://doi.org/10.1002/ctm2.702705‐methylcytosine modificationferroptosishepatocellular carcinomatranslational regulationYBX1
spellingShingle Ouwen Li
Ke An
Han Wang
Xianbin Li
Yueqin Wang
Lan Huang
Yue Du
Nuo Qin
Jiasheng Dong
Jingyao Wei
Ranran Sun
Yong Shi
Yanjia Guo
Xiangyi Sun
Ying Yang
Yun‐Gui Yang
Quancheng Kan
Xin Tian
Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
Clinical and Translational Medicine
5‐methylcytosine modification
ferroptosis
hepatocellular carcinoma
translational regulation
YBX1
title Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
title_full Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
title_fullStr Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
title_full_unstemmed Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
title_short Targeting YBX1‐m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
title_sort targeting ybx1 m5c mediates rnf115 mrna circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma
topic 5‐methylcytosine modification
ferroptosis
hepatocellular carcinoma
translational regulation
YBX1
url https://doi.org/10.1002/ctm2.70270
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