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Analysis of HBV Genomes Integrated into the Genomes of Human Hepatoma PLC/PRF/5 Cells by HBV Sequence Capture-Based Next-Generation Sequencing.
Genes (Basel). 2020 06 18; 11(6)G

Abstract

Hepatitis B virus (HBV) infection is a leading cause of hepatocellular carcinoma (HCC) worldwide. The integration of HBV genomic DNA into the host genome occurs randomly, early after infection, and is associated with hepatocarcinogenesis in HBV-infected patients. Therefore, it is important to analyze HBV genome integration. We analyzed HBV genome integration in human hepatoma PLC/PRF/5 cells by HBV sequence capture-based next-generation sequencing (NGS) methods. We confirmed the results by using Sanger sequencing methods. We observed that HBV genotype A is integrated into the genome of PLC/PRF/5 cells. HBV sequence capture-based NGS is useful for the analysis of HBV genome integrants and their locations in the human genome. Among the HBV genome integrants, we performed functional analysis and demonstrated the automatic expression of some HBV proteins encoded by HBV integrants from chromosomes 3 and 11 in Huh7 cells transfected with these DNA sequences. HBV sequence capture-based NGS may be a useful tool for the assessment of HBV genome integration into the human genome in clinical samples and suggests new strategies for hepatocarcinogenesis in HBV infection.

Authors+Show Affiliations

Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.Genome Medical Science Project, National Center for Global Health and Medicine, Ichikawa 272-8516, Japan.Genome Medical Science Project, National Center for Global Health and Medicine, Ichikawa 272-8516, Japan.Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32570699

Citation

Ishii, Tomotaka, et al. "Analysis of HBV Genomes Integrated Into the Genomes of Human Hepatoma PLC/PRF/5 Cells By HBV Sequence Capture-Based Next-Generation Sequencing." Genes, vol. 11, no. 6, 2020.
Ishii T, Tamura A, Shibata T, et al. Analysis of HBV Genomes Integrated into the Genomes of Human Hepatoma PLC/PRF/5 Cells by HBV Sequence Capture-Based Next-Generation Sequencing. Genes (Basel). 2020;11(6).
Ishii, T., Tamura, A., Shibata, T., Kuroda, K., Kanda, T., Sugiyama, M., Mizokami, M., & Moriyama, M. (2020). Analysis of HBV Genomes Integrated into the Genomes of Human Hepatoma PLC/PRF/5 Cells by HBV Sequence Capture-Based Next-Generation Sequencing. Genes, 11(6). https://doi.org/10.3390/genes11060661
Ishii T, et al. Analysis of HBV Genomes Integrated Into the Genomes of Human Hepatoma PLC/PRF/5 Cells By HBV Sequence Capture-Based Next-Generation Sequencing. Genes (Basel). 2020 06 18;11(6) PubMed PMID: 32570699.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of HBV Genomes Integrated into the Genomes of Human Hepatoma PLC/PRF/5 Cells by HBV Sequence Capture-Based Next-Generation Sequencing. AU - Ishii,Tomotaka, AU - Tamura,Akinori, AU - Shibata,Toshikatsu, AU - Kuroda,Kazumichi, AU - Kanda,Tatsuo, AU - Sugiyama,Masaya, AU - Mizokami,Masashi, AU - Moriyama,Mitsuhiko, Y1 - 2020/06/18/ PY - 2020/05/25/received PY - 2020/06/11/revised PY - 2020/06/16/accepted PY - 2020/6/24/entrez PY - 2020/6/24/pubmed PY - 2021/3/19/medline KW - HBV KW - HCC KW - Huh7 KW - PLC/PRF/5 KW - chromosome KW - hepatocarcinogenesis KW - integration KW - next-generation sequencing KW - transfection JF - Genes JO - Genes (Basel) VL - 11 IS - 6 N2 - Hepatitis B virus (HBV) infection is a leading cause of hepatocellular carcinoma (HCC) worldwide. The integration of HBV genomic DNA into the host genome occurs randomly, early after infection, and is associated with hepatocarcinogenesis in HBV-infected patients. Therefore, it is important to analyze HBV genome integration. We analyzed HBV genome integration in human hepatoma PLC/PRF/5 cells by HBV sequence capture-based next-generation sequencing (NGS) methods. We confirmed the results by using Sanger sequencing methods. We observed that HBV genotype A is integrated into the genome of PLC/PRF/5 cells. HBV sequence capture-based NGS is useful for the analysis of HBV genome integrants and their locations in the human genome. Among the HBV genome integrants, we performed functional analysis and demonstrated the automatic expression of some HBV proteins encoded by HBV integrants from chromosomes 3 and 11 in Huh7 cells transfected with these DNA sequences. HBV sequence capture-based NGS may be a useful tool for the assessment of HBV genome integration into the human genome in clinical samples and suggests new strategies for hepatocarcinogenesis in HBV infection. SN - 2073-4425 UR - https://www.unboundmedicine.com/medline/citation/32570699/Analysis_of_HBV_Genomes_Integrated_into_the_Genomes_of_Human_Hepatoma_PLC/PRF/5_Cells_by_HBV_Sequence_Capture_Based_Next_Generation_Sequencing_ DB - PRIME DP - Unbound Medicine ER -