抗HCV不同位点核酶细胞内对病毒基因的抑制作用  

INTRACELLULAR INHIBITION OF VIRAL GENE EXPRESSION BY HAMMERHEAD RIBOZYMES AGAINST HEPATITIS C VIRUS 5′ NCR AND C REGIONS

作　　者：焦成松[1] 周永兴[1] 贾战生[1] 冯志华 

机构地区：[1]第四军医大学唐都医院,西安710038

出　　处：《解放军医学杂志》2001年第7期471-473,共3页Medical Journal of Chinese People's Liberation Army

基　　金："九五"军队医药卫生科研基金资助课题 (编号 98D0 48)

摘　　要：为探讨核酶细胞内对HCV基因表达的抑制作用 ,笔者设计合成了两个针对HCV 5′NCR2 13位点和C区 498位点的锤头结构核酶 ,并插入真核表达载体 pcDNA3。应用WISH nc转基因细胞模型 ,通过脂质体法转染核酶表达载体 ,经发光检测仪测定荧虫素酶报告基因的表达变化以反应病毒基因的抑制率。结果显示 ,核酶2 13和核酶 498均能在细胞内降低荧虫素酶的表达 ,转染后 7日内的抑制率为 42 .94%～ 6 7.81%。提示增加核酶作用位点可防止病毒基因突变后的切割失效 。To investigate the potential use of synthetic stabilized ribozymes for the treatment of chronic hepatitis C virus (HCV) infection,we designed and synthesized 2 hammerhead ribozymes (Rz213 and Rz498) targeting conserved sites in the 5′noncoding region (NCR) and C gene of HCV RNA.Constructed to the eukaryotic vector pcDNA3, the two ribozymes were respectively or simultaneously transfected with lipofectamine into WISHnc transgenic cells, which could express permanently HCV C luciferase protein under the control of HCV 5′NCR.The expression of C luciferase was measured by luminometer.The results showed that the luciferase activities were significantly down regulated in the WISHnc cells, and the inhibitory rates were 42.94%～67.81% within 7 days after ribozymes transfection. There was no significant differences between Rz213 or Rz498 and co transfection, but adding the target site of ribozymes might prevent host cells from the loss of ribozyme therapeutic effect due to viral gene mutation.

关 键 词：C型肝炎样病毒属 核酶 荧虫素酶 丙型肝炎 抗HCV 

分 类 号：R512.63[医药卫生—内科学]