HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development

Abstract Background HBV integration is considered as the main contributor to hepatocellular carcinoma (HCC). However, whether HBV integrated sequences determine genotype pathogenicity and how to block their function during HCC progression remains unclear. Methods An in vitro HBV-infected PHH model a...

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Main Authors: Lu Chen, Wenxuan Li, Wenjing Zai, Xiangyi Zheng, Xianlong Meng, Qunyan Yao, Wei Li, Ying Liang, Mu Ye, Kaicheng Zhou, Mengxing Liu, Zhicong Yang, Zhanrui Mao, Hongyan Wei, Shuai Yang, Guoming Shi, Zhenghong Yuan, Wenqiang Yu
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Journal of Experimental & Clinical Cancer Research
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Online Access:https://doi.org/10.1186/s13046-025-03413-8
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author Lu Chen
Wenxuan Li
Wenjing Zai
Xiangyi Zheng
Xianlong Meng
Qunyan Yao
Wei Li
Ying Liang
Mu Ye
Kaicheng Zhou
Mengxing Liu
Zhicong Yang
Zhanrui Mao
Hongyan Wei
Shuai Yang
Guoming Shi
Zhenghong Yuan
Wenqiang Yu
author_facet Lu Chen
Wenxuan Li
Wenjing Zai
Xiangyi Zheng
Xianlong Meng
Qunyan Yao
Wei Li
Ying Liang
Mu Ye
Kaicheng Zhou
Mengxing Liu
Zhicong Yang
Zhanrui Mao
Hongyan Wei
Shuai Yang
Guoming Shi
Zhenghong Yuan
Wenqiang Yu
author_sort Lu Chen
collection DOAJ
description Abstract Background HBV integration is considered as the main contributor to hepatocellular carcinoma (HCC). However, whether HBV integrated sequences determine genotype pathogenicity and how to block their function during HCC progression remains unclear. Methods An in vitro HBV-infected PHH model and liver cancer cell lines were established to confirm the pathogenic potential of HBV-SITEs. The roles of HBV-SITE-1 in HCC development were analyzed using cellular phenotypic assays and molecular biology techniques, including the combined analysis of RNA-seq and ChIP-seq. Animal models were also used to evaluate the therapeutic effect of HBV-miR-2 inhibitors. Results We identified nine fragments of HBV Sequences Integrated To Enhancer, termed as “HBV-SITEs”. Particularly, a single nucleotide variation (T > G) was embedded at seed sequence of HBV-miR-2 in the highest integrated HBV-SITE-1 between genotypes B and H. Unexpectedly, B-HBV-SITE-1, not H-HBV-SITE-1, could abnormally activate oncogenic genes including TERT and accelerate HCC cell proliferation and migration. Meanwhile, HBV-miR-2 was gradually increased in HBV-infected cells and patient plasma with different HCC stages. Importantly, 227 genes upregulated by HBV, were also activated by HBV-miR-2 through triggering HBV-SITE-1 enhancer. Conversely, enhancer activities were particularly decreased by HBV-miR-2 inhibitors, and further downregulated activated oncogenic genes. Finally, HCC growth was dramatically restrained and HBV-induced transcripts were systematically reduced via injection of HBV-miR-2 inhibitors in animal models. Conclusion HBV-SITEs were identified as novel oncogenic elements for HCC, which provides an insightful perspective for the other cancers caused by oncogenic DNA viruses. We demonstrated that the integrated HBV sequence itself acted as oncogenic enhancers and nucleotide variations of HBV genotypes account for particular pathogenic progression, supporting that the viral nucleotide sequences are vital pathogenic substances beyond viral proteins. And modulation of their enhancer activities could be clinically achievable strategy for blocking DNA viruses-related cancer progression in the future.
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spelling doaj-art-af9c6baaec5946d49fd8561186d5e0b52025-08-20T03:08:40ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662025-05-0144112210.1186/s13046-025-03413-8HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma developmentLu Chen0Wenxuan Li1Wenjing Zai2Xiangyi Zheng3Xianlong Meng4Qunyan Yao5Wei Li6Ying Liang7Mu Ye8Kaicheng Zhou9Mengxing Liu10Zhicong Yang11Zhanrui Mao12Hongyan Wei13Shuai Yang14Guoming Shi15Zhenghong Yuan16Wenqiang Yu17Shanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Research Unit of Cure of Chronic Hepatitis B Virus Infection (CAMS), Shanghai Frontiers Science Center of Pathogenic Microbes and Infection, School of Basic Medical Sciences, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityDepartment of Liver Surgery and Transplantation, Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan UniversityDepartment of Liver Surgery and Transplantation, Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityPrecision Pharmacy and Drug Development Center, Department of Pharmacy, Tangdu Hospital, Fourth Military Medical UniversityDepartment of Liver Surgery and Transplantation, Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityDepartment of Liver Surgery and Transplantation, Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan UniversityKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Research Unit of Cure of Chronic Hepatitis B Virus Infection (CAMS), Shanghai Frontiers Science Center of Pathogenic Microbes and Infection, School of Basic Medical Sciences, Shanghai Medical College, Fudan UniversityShanghai Public Health Clinical Center & Laboratory of RNA Epigenetics, Institutes of Biomedical Sciences & Department of General Surgery, Huashan Hospital, Cancer Metastasis Institute, Shanghai Medical College, Fudan UniversityAbstract Background HBV integration is considered as the main contributor to hepatocellular carcinoma (HCC). However, whether HBV integrated sequences determine genotype pathogenicity and how to block their function during HCC progression remains unclear. Methods An in vitro HBV-infected PHH model and liver cancer cell lines were established to confirm the pathogenic potential of HBV-SITEs. The roles of HBV-SITE-1 in HCC development were analyzed using cellular phenotypic assays and molecular biology techniques, including the combined analysis of RNA-seq and ChIP-seq. Animal models were also used to evaluate the therapeutic effect of HBV-miR-2 inhibitors. Results We identified nine fragments of HBV Sequences Integrated To Enhancer, termed as “HBV-SITEs”. Particularly, a single nucleotide variation (T > G) was embedded at seed sequence of HBV-miR-2 in the highest integrated HBV-SITE-1 between genotypes B and H. Unexpectedly, B-HBV-SITE-1, not H-HBV-SITE-1, could abnormally activate oncogenic genes including TERT and accelerate HCC cell proliferation and migration. Meanwhile, HBV-miR-2 was gradually increased in HBV-infected cells and patient plasma with different HCC stages. Importantly, 227 genes upregulated by HBV, were also activated by HBV-miR-2 through triggering HBV-SITE-1 enhancer. Conversely, enhancer activities were particularly decreased by HBV-miR-2 inhibitors, and further downregulated activated oncogenic genes. Finally, HCC growth was dramatically restrained and HBV-induced transcripts were systematically reduced via injection of HBV-miR-2 inhibitors in animal models. Conclusion HBV-SITEs were identified as novel oncogenic elements for HCC, which provides an insightful perspective for the other cancers caused by oncogenic DNA viruses. We demonstrated that the integrated HBV sequence itself acted as oncogenic enhancers and nucleotide variations of HBV genotypes account for particular pathogenic progression, supporting that the viral nucleotide sequences are vital pathogenic substances beyond viral proteins. And modulation of their enhancer activities could be clinically achievable strategy for blocking DNA viruses-related cancer progression in the future.https://doi.org/10.1186/s13046-025-03413-8HBV Sequences Integrated To Enhancer (HBV-SITEs)Nuclear activating miRNAEnhancerHepatocellular carcinoma (HCC)HBV integration
spellingShingle Lu Chen
Wenxuan Li
Wenjing Zai
Xiangyi Zheng
Xianlong Meng
Qunyan Yao
Wei Li
Ying Liang
Mu Ye
Kaicheng Zhou
Mengxing Liu
Zhicong Yang
Zhanrui Mao
Hongyan Wei
Shuai Yang
Guoming Shi
Zhenghong Yuan
Wenqiang Yu
HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
Journal of Experimental & Clinical Cancer Research
HBV Sequences Integrated To Enhancer (HBV-SITEs)
Nuclear activating miRNA
Enhancer
Hepatocellular carcinoma (HCC)
HBV integration
title HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
title_full HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
title_fullStr HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
title_full_unstemmed HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
title_short HBV sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
title_sort hbv sequence integrated to enhancer acting as oncogenic driver epigenetically promotes hepatocellular carcinoma development
topic HBV Sequences Integrated To Enhancer (HBV-SITEs)
Nuclear activating miRNA
Enhancer
Hepatocellular carcinoma (HCC)
HBV integration
url https://doi.org/10.1186/s13046-025-03413-8
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