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  • Title: [Specific regulatory inhibition of transfected HepG(2) with HCV 5'NCR by antisense phosphorothioate oligodeoxynucleotides].
    Author: Wang S, Wang X, Guan W, Li M.
    Journal: Zhonghua Gan Zang Bing Za Zhi; 2000 Oct; 8(5):288-90. PubMed ID: 11058954.
    Abstract:
    OBJECTIVE: To screen efficient and specific new drugs against hepatitis C virus (HCV) and find the best target sequence of antisense oligodeoxynucleotides (ASODNs) targeting at HCV gene. METHODS: Fifteen S-ASODNs targeting at the 5'NCR and start AUG of HCV RNA were designed and synthesized according to the predicted RNA secondary structure of the HCV 5'NCR and the adjacent AUG region. HepG(2) cells were co-transfected with pHCV-neo4 and S-ASODN. The inhibitory effects of S-ASODNs on gene expression controlled by HCV 5'NCR were determined by the assay of luciferase activity. RESULTS: Five S-ASODNs, i.e. HCV65, HCV279, HCV363, HCV349 and HCV352, showed sequence-specific and dose-dependent inhibitory activities with an inhibition rate of more than 80% at a concentration of 100 nmol/L. According to 50% inhibitory concentrations, the inhibitory activity of HCV363 was the best. On the other hand, ASODNs had only little non-specific effect by negative control trials. In addition, the results also indicated that there were some coordinate effects between ASODNs at different targets. CONCLUSION: Stem-loop 2a and 3d of HCV 5'NCR and the start AUG of polyprotein precursor are candidate targets of S-ASODNs.
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