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Objective To study the effect and mechanism of antisense oligodeoxynucleotides (ASONs) inhibiting hepatitis B virus (HBV) expression. Methods We designed and synthesized antisense oligodeoxynucleotides directly against HBV PreS 2 gene and noncomplementary sequence control. 2.2.15 cells were chosen as cell model. Inhibitory effect of ASONs on HBV gene expression were assayed by ELISA. Cell apoptosis and proliferation were detected by Fascan Flow Cell Cytometer. Effect of ASONs on cell metabolism was detected by radioimmunoassay (RIA) and MTT assay. Results ASONs were able to effectively inhibit HBV expression. Their inhibitory rates of HBsAg and HBeAg were 66% and 91%, respectively. Noncomplementary sequence control group (both inhibitory rates were 11%) was not able to inhibit HBV expression. ASON might induce host cell apoptosis. Cell apoptosis rates on 3rd and 6th day were 6.10% and 6.43%, respectively. Proliferation index were 37% and 36%, respectively. Results of RIA and MTT showed that ASON had not cytoxicity on host cell. Conclusions Not only are ASON able to inhibit HBV gene expression with sequence specific but also clear HBV in the way of apoptosis.
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0.05).The C myc and N ras protein expression of 2.2.15 were reduced after ASONs were used 3 days.There was a significant difference between ASON group and control group ( P
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Objective:To screen the high performance,specific and nontoxic anti HBV antisense oligonucleotide fragment.Methods:Taking 2.2.15 cells as cell model,the antisense oligodeoxynucleotides(ASON)toward the HBV regulation genesenhancerⅡ(HBV EFⅡ)were designed and synthesized.The role played by ASON in inhibiting the secretion and expession of HBsAg and HBeAg by the host cells was detected by ELISA method.The effect of ASON on prolifezation and metabolism of the cells was detected by MTT method.Results:The inhibitory rate of HBsAg by ASON was 92% and 75%,respectively,indicating that itsdifference from that of noncomplementary sequence control group(inhibitory rate was 11%)had considerable statistical significance( P 0.05).Conclusion:ENⅡ is one of the most important target oriented sequencial selective regions of research on anti HBV nature of antisense oligonucleotide.