硫代反义寡核苷酸对裸鼠异植瘤HCV 5′NCR基因表达的抑制作用

目的 在HCV 5′NCR转基因细胞模型HepG2.9706细胞建立成功的基础上,采用裸鼠异植瘤模型进行HCV特异性硫代反义寡核苷酸（HCV363、HCV279及HCV349）的体内活性评价。方法 将HepG2.9706细胞接种于4－6周龄的雌性BALB／C裸鼠的侧腹皮下,动物随机分组,约1周后,采用腹腔注射途径,开始给药。结果 针对HCV 5′NCR的硫代反义寡核苷酸HCV363、HCV279及HCV349对异植瘤细胞内HCV 5′NCR调控荧光素酶表达具有明显的序列特异性抑制作用,抑制率分别为80．4％、78．6％及47．9％。三个不同浓度的HCV363作用结果表明随着药物浓度的增加,HCV363对荧光素酶基因的表达抑制活性明显增强,最大抑制率可达82．7％。结论 该研究提示反义寡核苷酸有可能成为HCV感染治疗的新途径。

Inhibitory effects of phosphorothioate antisense oligonucleotides on gene expression controlled by HCV 5'NCR in nude mice xenograft

OBJECTIVE: To evaluate the activity in vivo of 3 phosphorothioate antisense oligonucleotides, HCV363, HCV279 and HCV349 using nude mice xenograft models based on the establishment of HCV 5' NCR transgenic cellular model (HepG2.9706). METHODS: Female BALB/C nude mice, aged 4 to 6 weeks and weighing around 20g, were implanted s.c. with 100mul (106 cells) of the HepG2.9706 cells suspension in the lower-back region. In approximate 1 weeks, the animals were randomly grouped and intraperitoneally administrated the antisense drugs at 10 mg/kg body weight. RESULTS: HCV363, HCV279 and HCV349 had obvious sequence-specific inhibitory effects on luciferase expression controlled by HCV 5' NCR in xenograft cells with the inhibitory rates of 80.4%, 78.6% and 47.9%, respectively. The effects of three different concentrations (5, 10, 20 mg/kg body weight) of HCV363 indicated that HCV363 increased the inhibitory activities on luciferase expression following the concentration raise and its inhibitory rate was up to 82.7%. CONCLUSIONS: This study suggests that antisense oligonucleotides may provide a novel therapeutic approach to the treatment of HCV infection.