[A study of hepatitis B virus(HBV) anti-genome and its inhibitory effect on HBV replication].
OBJECTIVE
To study the combination of triplex forming oligodeoxynucleotides (TFO) with hepatitis B virus gene and its effect on replication of HBV.


METHODS
An oligodeoxynucleotide (TFO20) which can form triplex with SP1 sites in HBV core promoter was synthesized and was linked with biotin. Both the liposome-TFO20 and naked TFO20 were transfected to HepG2.2.15 cells, which was detected by SABC assay and showed signals that TFO20 was combined with HBV DNA. ELISA, reverse transcript (RT)-PCR and fluorescence quantitative PCR were used to investigate the change of HBV DNA, 3.5 kb/3.4 kb RNA, 2.4 kb/2.1 kb RNA, HBsAg and HBeAg in HepG2.2.15 cells before and after treatment with TFO20.


RESULTS
The ratio of liposome-TFO20 and naked TFO20 transfected to HepG2.2.15 cells were 80% and 40%, respectively. TFO20 signals mainly distributed in the nucleus and cytoplasm. After treatment with TFO20, the level of HBV DNA was reduced from 10(7.01) to 10(4.94), while 3.5 kb/3.4 kb RNA, 2.4 kb/2.1 kb RNA, HBsAg and HBeAg were also reduced by 70.2%, 31.6%, 37.0%, 78.2% respectively.


CONCLUSIONS
TFO20 can be transfected into cells by packing with liposome and can be combined with HBV DNA. TFO20 can effectively inhibit replication of HBV.
Gao
Y
Y
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Central China Science and Technology University, Wuhan 430022, China.
Luo
D
D
Cai
S
S
Zeng
L
L
Li
S
S
D004267:DNA Viruses / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication / Q000235:genetics*
D020319:Oligodeoxyribonucleotides, Antisense