靶向c-Raf-1基因的反义核酸抗乙型肝炎病毒活性研究  

Inhibition of Hepatitis B Virus Gene Expression by Antisense Oligo nucleotide Targeting c-Raf-1 Gene

作　　者：黄海[1,2] 张秀娟[2] 谢佩雯[2] 王学军[2] 王升启[2] 

机构地区：[1]河南中医学院,河南郑州450003 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《生物技术通讯》2013年第5期616-620,共5页Letters in Biotechnology

基　　金：国家"十二五"重大新药创制专项(2012ZX09103301-044)

摘　　要：目的:通过体内外实验验证靶向c-Raf-1基因的反义核酸是否具有抑制乙型肝炎病毒(HBV)的活性。方法:设计靶向c-Raf-1基因的反义核酸,并在细胞水平进行体外抗HBV活性筛选,通过RT-PCR检测c-Raf-1基因mRNA水平的变化,通过体内药效学实验进一步验证反义核酸的抗HBV效果。结果:经体外筛选,靶向c-Raf-1基因的反义核酸Raf-3145具有相对明显的抑制HBV表面抗原(HBsAg)的作用,并可剂量依赖性地抑制c-Raf-1基因的表达;体内药效学结果显示,反义核酸Raf-3145在30 mg/kg剂量下对HBsAg的表达具有一定的抑制作用。结论:经体内外活性评价,初步确定了靶向宿主基因c-Raf-1的反义核酸具有一定的抑制HBsAg表达的活性,也进一步验证了c-Raf-1基因可以作为抗HBV药物设计的候选靶点。Objective： To evaluate the activity of antisense oligonucleotide targeting c-Raf-1 gene for hepatitis B virus（HBV） inhibition both in vitro and in vivo. Methods： The antisense oligonucleotides targeting c-Raf-1 were designed and screened for their anti-HBV activity in HepG2.2.15 cells. Subsequently, the mRNA expression level of c-Raf-1 was detected by RT-PCR assay. Furthermore, the anti-HBV effect of antisense oligonucleotide was evaluated in vivo using an hydrodynamic injection mouse model for HBV. Results： The antisense oligonucle-otide Raf-3145 inhibited hepatitis B surface protein（HBsAg） expression significantly in vitro and the c-Raf-1 mRNA expression was also down-regulated dose-dipendently by it; the in vivo experiment showed that Raf-3145 could also inhibited HBsAg expression at the dose of 30 mg/kg. Conclusion： The antisense oligonucleotide Raf-3145 targeting c-Raf-1 could inhibited HBsAg expression both in vitro and in vivo, and the results also dem-onstrated that the c-Raf-1 gene could be as a potential target for anti-HBV drug design in future.

关 键 词：乙型肝炎病毒 反义核酸 c-Raf-1基因 

分 类 号：Q78[生物学—分子生物学]