Comparison of the inhibiting effects of four antisense oligedeoxynucleotide
phosphorothioates on hepatitis B virus gene expression

YUAN Zheng-Hong; HE Li-Fang; ZHAO Su-Ling; WU Xiang-Fu; CHEN Chang-Qiang

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February 1995

YUAN Zheng-Hong, HE Li-Fang, ZHAO Su-Ling, WU Xiang-Fu and CHEN Chang-Qiang. Comparison of the inhibiting effects of four antisense oligedeoxynucleotidephosphorothioates on hepatitis B virus gene expression[J]. Virologica Sinica, 1995, 10(1).

Citation: YUAN Zheng-Hong, HE Li-Fang, ZHAO Su-Ling, WU Xiang-Fu, CHEN Chang-Qiang. Comparison of the inhibiting effects of four antisense oligedeoxynucleotide phosphorothioates on hepatitis B virus gene expression .VIROLOGICA SINICA, 1995, 10(1) : 34.

反义寡聚脱氧核苷险(ODN)衍生物抑制乙肝病毒抗原表达的研究

1.
上海医科大学医学分子病毒学实验室中国科学院上海生物化学研究所中国科学院上海生物工程研究中心

摘要

为比较针对中不同基因的ＯＤＮ硫代衍生物阻断乙型肝炎病毒（ＨＢＶ）的抗原表达，合成了与核心蛋白编码基因起始码上游序列（＃１８９３－１９０７），多聚酶蛋白编码基因起始码上游及内部序列（＃２２９７－２３１３，＃２７９７－２８１３）３．５ｋｂＲＮＡ３′端起始负链ＤＮＡ合成序列（＃ｌ８１７－１８３１）互补的寡聚脱氧核苷酸硫代衍生物［Ｃ－ＯＤＮ，Ｐ－ＯＤＮ，ＰＭ－ＯＤＮ，ＰＢＳ－ＯＤＮ］，分别在ＨＢＶ短暂表达和稳定表达细胞培养系统中观察其抑制ＨＢＶ抗原表达的作用。结果发现，当培养液中ＯＤＮ浓度为２０μｍｏｌ／Ｌ时，四种ＯＤＮ均能抑制ＨｅｐＧ２２．２．１５细胞ＨＢｓＡｇ（乙肝表面抗原）的产生（抑制率分别为：Ｃ－ＯＤＮ，２４．５％；ＰＯＤＮ，１３．４％；ＰＭ－ＯＤＮ，３８．４％；ＰＢＳ－ＯＤＮ，２８．４％）；与ＨＢＶＤＮＡ双体质粒共转染ＨｅｐＧ２细胞后部分ＯＤＮ可明显抑制ＨＢＶＤＮＡ在细胞内表达（ＨＢｅａｇ分泌抑制率分别为：Ｃ－ＯＤＮ，５２％；ＰＭ－ＯＤＮ，７０％；Ｐ－ＯＤＮ，４８％；ＰＢＳ－ＯＤＮ，１８％）。用测定上清中人白蛋白的结果证明，Ｃ－ＯＤＮ的细胞毒作用最低。对Ｃ－ＯＤＮ的作用机理及进一步发展ＯＤＮ的应用也进行了讨论。
- 乙型肝炎病毒反义寡聚脱氧核苷酸硫代衍生物基因表达抑制

Comparison of the inhibiting effects of four antisense oligedeoxynucleotide phosphorothioates on hepatitis B virus gene expression

1.
Shanghai Medical University

Abstract

In this study we evaluated the effects of four antisense oligodeoxynucl eotide phosphorothioates(ODN)asainst antigen expression of HBV gene,Four ODNs were synthesized with an average length of fifteen bases directed against the region upstream of the core gene initiation cedon(#1893-1907),against the region upstream and downstream of the polymerase codin8 gene initiation codon(#2297-2313,# 2797-2813),against the 3 region of 3.5 kb RNA for initiation of reverse transcription(#1817-l831).An ODN with random sequences was also synthesized as control.ODNs were tested agair1st HBV in both HBV transient and stable expressing systems.When being added to epG2 2.2.15 cells at 20 μmol/L every other day,compared to control,all our ODNs showed variable ex tent of inhibition on the expression of HBsAg.The inhibiting rates at day 4 were:C-ODN,24.5%;P-ODN,13%.PM-ODN,38.4%;PBS-ODN,28.4%,However all ODNs had no effect on the ex pression of HBeAg in th is cell line,After being cotransfected into the HepG2 cells with HBV DNA dime
- epatitis B virus Antisense oligodeoxynucleotide phosphorothioate Inhibition of gene ex pression

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Comparison of the inhibiting effects of four antisense oligedeoxynucleotide phosphorothioates on hepatitis B virus gene expression

1. Shanghai Medical University

Abstract: In this study we evaluated the effects of four antisense oligodeoxynucl eotide phosphorothioates(ODN)asainst antigen expression of HBV gene,Four ODNs were synthesized with an average length of fifteen bases directed against the region upstream of the core gene initiation cedon(#1893-1907),against the region upstream and downstream of the polymerase codin8 gene initiation codon(#2297-2313,# 2797-2813),against the 3 region of 3.5 kb RNA for initiation of reverse transcription(#1817-l831).An ODN with random sequences was also synthesized as control.ODNs were tested agair1st HBV in both HBV transient and stable expressing systems.When being added to epG2 2.2.15 cells at 20 μmol/L every other day,compared to control,all our ODNs showed variable ex tent of inhibition on the expression of HBsAg.The inhibiting rates at day 4 were:C-ODN,24.5%;P-ODN,13%.PM-ODN,38.4%;PBS-ODN,28.4%,However all ODNs had no effect on the ex pression of HBeAg in th is cell line,After being cotransfected into the HepG2 cells with HBV DNA dime