title: Specific inhibition of hepatitis b virus gene expression by an antisense oligonucleotide in vitro.
abstract: It was previously shown that a number of antisense oligonucleotides against hepatitis B virus (HBV) mRNAs were highly effective in inhibition of HBV gene expression (Yao et al., 1995). Here, using radioisotope techniques, we report a specific inhibition of HBV surface antigen (HBsAg) production in vitro by 2.2.15 cells (Hep-G2 cells transfected with HBV genome) by the antisense oligonucleotide 15-S-asON, a 15-mer phosphorothioate analogue complementary to the cap site of the SPII promoter of HBV mRNA, at a concentration of 2-5 mumol/l. After 24 and 48 hrs of incubation of cells with 15-S-asON, the intracellular concentration of the latter rose to 69.4 and 75.8 nmol/l, respectively, and the HBsAg level assayed by ELISA was reduced by 50.0% and 70.6%, respectively. These results were checked by use of the radioimmunoprecipitation method: 2.2.15 cells exposed to 15-S-asON and labelled with [35S]-methionine for 48 hrs showed a decrease of the HBsAg level by 81.26% but almost none of the total proteins. No cytotoxicity of the 15-S-asON was observed with regard to the cell morphology and growth. These results indicate that the tested antisense oligonucleotide specifically inhibits the HBV gene expression.
authors.affiliation: Department of Infectious Diseases, Fourth Military Medical University, Xi'an, P.R. of China.
authors.forename: Z Q
authors.initials: ZQ
authors.lastname: Yao
authors.forename: Y X
authors.initials: YX
authors.lastname: Zhou
authors.forename: X M
authors.initials: XM
authors.lastname: Feng
authors.forename: Z X
authors.initials: ZX
authors.lastname: Chen
mesh_terms: D002460:Cell Line; D002470:Cell Survival; D015967:Gene Expression Regulation, Viral / Q000187:drug effects* / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000187:drug effects* / Q000276:immunology; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012314:RNA Cap Analogs / Q000494:pharmacology*
chemical_list: D006514:Hepatitis B Surface Antigens; D016376:Oligonucleotides, Antisense; D012314:RNA Cap Analogs