HepG2.9706细胞内HDV核酶对HCV RNA抑制活性的初步研究  

Preliminary studies on the intracellular inhibiting activity of HDV ribozymes against HCV RNA

作　　者：郭焕珍[1] 毛青[1] 李奇芬[1] 王宇明[1] 顾长海[1] 于乐成[1] 

机构地区：[1]第三军医大学西南医院全军感染病研究所,重庆400038

出　　处：《第三军医大学学报》2004年第11期960-962,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 396 0 0 0 31)~~

摘　　要：目的　探讨HDV核酶在细胞内抑制HCVRNA的可能性。方法　将重组载体转染至转基因细胞HepG2 .970 6中 ,通过荧光定量PCR检测细胞和培养上清中的HCVRNA ,初步探讨HDV核酶对HCVRNA的抑制活性。结果　①RzC1,RzC2 ,RzC3 3组HDV核酶定向插入到真核表达载体。②在转基因细胞HepG2 .970 6中 ,pC1 RzC1、pC1 RzC2、pC1 RzC3对HCV 5′NCR CRNA抑制活性分别为 69.2 %,3 8.7%、4.2 %。结论　①成功构建了 3个HDV核酶重组表达载体。②pC1 RzC1、pC1 RzC2在转基因细胞HepG2 .970 6具有抑制HCV 5′NCRObjective To discuss the possibility of hepatitis delta virus (HDV) ribozymes inhibiting HCV RNA in cells. Methods Three recombined expression vectors were transfected to engineered cell HepG2.9706 using liposome mediated method. The inhibitory effects of HDV were evaluated by HCV RNA quantitation in cultured cells and supernatants. Results ① The mammalian expression vectors contained ribozymes cDNA of RzC1, RzC2, and RzC3. ② The inhibitory effects of pC1 RzC1, pC1 RzC2, and pC1 RzC3 were 69.2%, 38.7% and 4.2%, respectively, in the HepG2.9706. Conclusion ① Three eukaryotic expression vectors are constructed. ② The inhibitory activity of pC1 RzC1 (107-113 nt) and pC1 RzC2 (268-274 nt) is more efficient than that of pC1 RzC3 (345-351 nt) in HepG2.9706 cells.

关 键 词：核酶 HDV核酶 丙型肝炎 荧光定量PCR 

分 类 号：R394.3[医药卫生—医学遗传学] R512.6[医药卫生—基础医学]