Dynamics of HBV cccDNA expression and transcription in different cell growth phase

Journal of Biomedical Science

Research

Mong-Liang Chen¹  Cheng-po Hu²  Chungming Chang³  Chien-Chiao Huang³  Chin-Liew Chong³  King-Song Jeng⁴  Yu-Chi Chou⁴  Yi-Chieh Wu⁵  Kuen-Nan Tsai⁶ 

[1] Center for Molecular Medicine, China Medical University and Hospital, 2, Yude Road, Taichung, 40447, Taiwan;Department of Life Science, Tunghai University, 181, Sec.3, Taichung Port Road, 40704, Taichung, Taiwan;Institute of Microbiology and Immunology, National Yang-Ming University, 155, Sec.2, Linong Street, 112, Taipei, Taiwan;Institute of Molecular and Genomic Medicine, National Health Research Institutes, 35, Keyan Road, 350, Zhunan Town, Miaoli, Taiwan;Institute of Molecular Biology, Academia Sinica, 128, Sec. 2, Academia Road, Nankang, 115, Taipei, Taiwan;Institute of Molecular and Genomic Medicine, National Health Research Institutes, 35, Keyan Road, 350, Zhunan Town, Miaoli, Taiwan;Institute of Molecular and Genomic Medicine, National Health Research Institutes, 35, Keyan Road, 350, Zhunan Town, Miaoli, Taiwan;Institute of Molecular Medicine, National Tsing Hua University, 101, Sec.2, Kuang-Fu Road, 30013, Hsinchu, Taiwan;

关键词: HBV;    cccDNA;    viral replication;    cell proliferation;    growth confluency;   

DOI  :  10.1186/1423-0127-18-96

 received in 2011-11-18, accepted in 2011-12-30,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThe covalently closed-circular DNA (cccDNA) of hepatitis B virus (HBV) is associated with viral persistence in HBV-infected hepatocytes. However, the regulation of cccDNA and its transcription in the host cells at different growth stages is not well understood.MethodsWe took advantages of a stably HBV-producing cell line, 1.3ES2, and examine the dynamic changes of HBV cccDNA, viral transcripts, and viral replication intermediates in different cellular growth stages.ResultsIn this study, we showed that cccDNA increased suddenly in the initial proliferation phase of cell growth, probably attributable to its nuclear replenishment by intracellular nucleocapsids. The amount of cccDNA then decreased dramatically in the cells during their exponential proliferation similar to the loss of extrachromosomal plasmid DNA during cell division, after which it accumulated gradually while the host cells grew to confluency. We found that cccDNA was reduced in dividing cells and could be removed when proliferating cells were subjected to long term of lamivudine (3TC) treatment. The amounts of viral replicative intermediates were rapidly reduced in these proliferating cells and were significantly increased after cells reaching confluency. The expression levels of viral transcripts were increased in parallel with the elevated expression of hepatic transcription factors (HNF4α, CEBPα, PPARα, etc.) during cell growth confluency. The HBV transcripts were transcribed from both integrated viral genome and cccDNA, however the transcriptional abilities of cccDNA was less efficient then that from integrated viral genome in all cell growth stages. We also noted increases in the accumulation of intracellular viral particles and the secretion of mature virions as the cells reached confluency and ceased to grow.ConclusionsBased on the dynamics of HBV replication, we propose that HBV replication is modulated differently in the different stages of cell growth, and can be divided into three phases (initial proliferation phase, exponential proliferation phase and growth confluency phase) according to the cell growth curve. The regulation of cccDNA in different cell growth phase and its importance regarding HBV replication are discussed.

【 授权许可】

Unknown   
© Chong et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.