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目的建立HCV感染动物模型,并对HCV进行RNA干涉。方法制备HCV感染的HepG2细胞,RT-PCR法鉴定后腋下注射裸鼠,免疫组化法检测瘤组织内HCV NS3表达情况。制备带绿色荧光蛋白(GFP)的HCVsiRNA表达质粒,转染HepG2细胞,观察GFP表达情况。将此质粒注射模型小鼠,取瘤组织,RT-PCR及免疫组化法对其进行检测。结果在HCV感染的裸鼠实体瘤中检测到HCV NS3蛋白的稳定表达。成功构建了HCV发夹siRNA表达质粒,该质粒转染细胞后能够在其内正确表达。将该质粒尾静脉注射HCV感染的模型小鼠,注射4d后瘤组织内HCV NS3抗原阳性细胞阴转。结论成功建立了HCV感染的裸鼠模型,构建了HCV发夹siRNA表达质粒,该质粒能成功地干涉模型小鼠中的HCV。
Objective To establish an animal model of HCV infection and perform RNA interference on HCV. Methods HCV infected HepG2 cells were prepared and injected into nude mice by RT-PCR. Immunohistochemistry was used to detect the expression of HCV NS3 in the tumor tissue. HCV siRNA expression plasmid with green fluorescent protein (GFP) was prepared and transfected into HepG2 cells to observe GFP expression. The plasmid was injected into the model mice to detect the tumor tissue, RT-PCR and immunohistochemistry. Results The stable expression of HCV NS3 protein was detected in HCV-infected solid tumor of nude mice. The HCV hairpin siRNA expression plasmid was successfully constructed and transfected into the cells and correctly expressed in the cells. The plasmid was injected intravenously into the tail vein of HCV-infected model mice, and the HCV NS3 antigen-positive cells in the tumor tissue were negatively transfected 4 days after injection. Conclusion The HCV infected nude mouse model was successfully established and the HCV hairpin siRNA expression plasmid was successfully constructed and successfully interfered with the HCV in model mice.