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[A study of hepatitis B virus(HBV) anti-genome and its inhibitory effect on HBV replication]
1Department of Infectious Diseases, Union Hospital, Tongji Medical College, Central China Science and Technology University, Wuhan 430022, China.
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.