摘要
为了观察逆转录病毒载体包装细胞系统介导反义基因转移表达的抗乙型肝炎病毒（HBV）作用，将HBVayw前C／C基因（preC／C）片段反向插入逆转录病毒载体质粒。将重组体转染逆转录病毒包装细胞PA317后，获得重组逆转录病毒。用重组逆转录病毒感染2．2．15细胞后发现，感染后第3天，细胞培养上清HBV表面抗原（HBsAg）和e抗原（HBeAg）表达量即明显减少，抑制作用于感染后第5天达到高峰，其中对HBsAg表达的抑制率为27．0％，对HBeAg表达的抑制率为59.5％。反义基因重组逆转录病毒感染2.2．15细胞对HBV抗原表达的抑制作用至少可以持续到转导后的第11天。空载及正义基因重组逆转录病毒感染对2．215细胞HBV抗原表达无明显抑制作用。此外，反义基因重组逆转录病毒感染对2.2.15细胞HBVDNA复制也有抑制作用．无细胞毒性。
Retroviral vector expresing antisense RNA complementary to HBV pre C/C (ANTI - C) was used to transduce human hepatoblastoma cell 2. 2. 15, which was transfected with HBV DNA and can express HBV markers. The results showed that the inhibitory effects of antisense gene transfer mediated by retroviral vector ANTI - C on the expression of HBV antigens appeared as early as on the 3rd day after transduction, reached peak level on the fifth day, and persisted at least for 11 days. The inhibitory rates of HBsAg and HBeAg in 2. 2. 15 cells by ANTI - C were 27. 0 %and 59. 5 % on the fifth day after transduction, and in comparison to blank or sense - expressing vectors, the inhibitory effects of ANTI - C were highly significant (P＜0.01 ). HBV DNA in the supernatants of 2. 2. 15 cells transduced with ANTl - C in comparison to blank or SENSE - C was also reduced on the fifth day after transduction as detected by DNA dot blot assays, but the viability of transduced 2. 2. 15 cells was not affected as indicated by MTT assays. Our results demostrated that HBV can be inhibited by retroviral vectors containing antisense gene and the antisense retroviral vectors may be potentially useful for anti - HBV gene therapy.
出处
《中国病毒学》
CSCD
1997年第3期214-218,共5页
Virologica Sinica
基金
国家自然科学基金
