Effects of two novel nucleoside analogues on different hepatitis B virus promoters.
OBJECTIVE
To explore the effects of the nucleoside analogues beta-L-D4A and beta-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 beta-L-D4A and beta-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 beta-L-D4A in a dose-dependent manner, while expression of EGFP under the control of the core promoter (Cp) and Xp was inhibited by beta-LPA in a dose-dependent manner.


CONCLUSIONS
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.
He
Xing-Xing
XX
Institute of Liver Diseases, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Lin
Ju-Sheng
JS
Chang
Ying
Y
Zhang
Ying-Hui
YH
Li
Yan
Y
Wang
Xiao-Yan
XY
Xu
Dong
D
Cheng
Xiao-Ming
XM
D015075:2-Aminopurine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D016048:Dideoxyadenosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D015394:Molecular Structure; D011401:Promoter Regions, Genetic
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D015075:2-Aminopurine; D016048:Dideoxyadenosine; C075923:2'3'-didehydro-2'3'-dideoxyadenosine