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Effects of two novel nucleoside analogues on different hepatitis B virus promoters

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Author(s): Xing-Xing He, Ju-Sheng Lin, Ying Chang, Ying-Hui Zhang, Yan Li, Xiao-Yan Wang, Dong Xu, Xiao-Ming Cheng

Journal: World Journal of Gastroenterology
ISSN 1007-9327

Volume: 14;
Issue: 12;
Start page: 1836;
Date: 2008;
Original page

Keywords: Hepatitis B virus | Nucleoside analogue | Hepatitis B virus promoter | Enhanced green fluorescent protein | Fluorescence activated cell sorter

ABSTRACT
AIM: To explore the effects of the nucleoside analogues β-L-D4A and β-LPA on hepatitis B virus (HBV) promoters.METHODS: Four HBV promoters were amplified by polymerase chain reaction (PCR) and subcloned into the expression vector pEGFP-1. The four recombinants controlled by HBV promoters were confirmed by restriction analysis and sequencing. Human hepatoma HepG2 cells transfected with the recombinant plasmids were treated with various concentrations of β-L-D4A and β-LPA. Then, enhanced green fluorescent protein (EGFP)-positive cells were detected by fluorescence microscopy and using a fluorescence activated cell sorter (FACS).RESULTS: Four HBV promoters were separately obtained and successfully cloned into pEGFP-1. Expression of EGFP under the control of the surface promoter (Sp) and the X promoter (Xp) was inhibited by β-L-D4A in a dose-dependent manner, while expression of EGFP under the control of the core promoter (Cp) and Xp was inhibited by β-LPA in a dose-dependent manner.CONCLUSION: The two novel nucleoside analogues investigated here can inhibit the activities of HBV promoters in a dose-dependent manner. These findings may explain the mechanisms of action by which these two novel compounds inhibit HBV DNA replication.
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