硫代反义寡聚脱氧核苷酸体外对乙型肝炎病毒基因表达的抑制作用  

The inhibition of hepatitis B viral gene expression by antisense oligodeoxynucleotides

作　　者：吴庆[1] 张胜权[2] 张在和[2] 

机构地区：[1]安徽医科大学第一附属医院检验科,合肥230022 [2]安徽医科大学生化教研室,合肥230022

出　　处：《安徽医科大学学报》2001年第6期434-437,共4页Acta Universitatis Medicinalis Anhui

摘　　要：目的　探讨乙型肝炎病毒 (HBV)C基因翻译起始区和C基因区硫代反义寡聚脱氧核苷酸 (ASON)对HBV表达的序列性抑制作用。方法　以 2 2 15细胞系作为实验对象 ,设计了一系列互补于HBVmRNA编码核心蛋白基因区(C区 ) 15聚硫代ASON ,通过脂质体介导将这些合成的外源基因导入 2 2 15细胞中 ,用酶联免疫吸附法 (ELISA)检测作用细胞培养上清液中乙型肝炎病毒E抗原 (HBeAg)和乙型肝炎病毒表面抗原 (HBsAg)含量。 结果　与HBVmRNAC基因起始区互补的硫代ASON序列特异性地抑制乙型肝炎病毒基因表达 ,而正义硫代寡聚脱氧核苷酸未见有抑制效应 ,与HBVmRNAC基因内部互补的序列很少或无抗病毒活性。另外 ,硫代ASON对细胞生长无毒性作用。结论　硫2 0 0 1 0 7 0 3收稿 ,2 0 0 1 10 15修回作者单位 :1 安徽医科大学第一附属医院检验科 ,合肥　 2 30 0 2 22 安徽医科大学生化教研室 ,合肥　 2 30 0 2 2作者简介 :吴　庆 ,女 ,36岁 ,主管技师 ,硕士研究生 ,主攻方向 :生物化学张在和 ,女 ,6 2岁 ,教授 ,硕士生导师 ,研究方向Objective To explore the inhibition of hepatitis B viral gene expression in 2.2.15 cells in vitro by antisense oligodeoxynucleotides. Methods The HepG2 cells transfected with HBV DNA were chosen as the target cells and a series of antisense phosphorothioate oligodeoxynucleotides (ASON),complementary to core regions of HBV gene were synthesized. This was accomplished through the use of lipofectin transfection reagent which extracelluar ODNs were transfectected into the 2.2.15 cells. At intervals, medium was sampled and assayed for hepatitis B surface antigen (HBsAg)and hepatitis e antigen (HBeAg) by ELISA. Results ASONs directed against the initiator of gene C of HBV mRNA were found to inhibit specifically gene expression of HBV. Neither the same sense oligomers nor the antisense ODN directed against the middle region of gene C had such effect on viral antigen production.All of the oligonucleotides at the concentration tested had no cytotoxicity. Conclusion The results suggest a therapeutic potential for ASON in the treatment of chronic HBV patients.

关 键 词：乙型肝炎病毒 反义寡核苷酸类 基因表达 

分 类 号：R512.62[医药卫生—内科学] R373.21[医药卫生—临床医学]