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目的:构建表达新城疫病毒(newcastle disease virus,NDV)HN基因的重组鸡痘病毒vFVHN,探讨vFVHN对体外培养人肝癌细胞SMMC7721的抑制作用和对小鼠荷肝癌模型的体内抑瘤效果。方法:以野生型鸡痘病毒(fowl poxvirus,FPV)282 E4株为载体,构建表达新城疫病毒HN基因的重组鸡痘病毒vFVHN。采用5-溴-2-脱氧尿苷(5-bromo-2-deoxyribouri- dine,BrdU)加压法筛选重组体,并通过RT-PCR和Western blot等方法对其进行鉴定。采用噻唑蓝(methylthiazoletetrazolium,MTT)染色法检测vFVHN对人肝癌细胞SMMC7721的杀伤率,并通过检测抑瘤率观察其在C57BL/6小鼠荷肝癌模型的抑瘤效果。结果:成功构建重组鸡痘病毒vFVHN,其对SMMC7721细胞的杀伤率为43.37%,对C57BL/6小鼠肝癌模型的抑瘤率为20.65%。结论:重组鸡痘病毒vFVHN可有效杀伤体外培养的人肝癌细胞SMMC7721,并对C57BL/6小鼠荷肝癌模型体内的实体肿瘤有一定抑制作用。
OBJECTIVE: To construct recombinant fowlpox virus vFVHN expressing HN gene of newcastle disease virus (NDV) and investigate the inhibitory effect of vFVHN on human hepatocellular carcinoma cell line SMMC7721 in vitro and its anti-tumor effect in vivo. Methods: The recombinant fowlpox virus vFVHN expressing Newcastle disease virus (HN) gene was constructed by using the wild-type fowl poxvirus strain 282 E4. The recombinant was screened by 5-bromo-2-deoxyribouri-dine (BrdU) pressurization and identified by RT-PCR and Western blot. The killing rate of vFVHN on SMMC7721 cells was detected by methyl thiazoletetrazolium (MTT) staining. The inhibitory effect of vFVHN on the inhibition of SMMC7721 cells was observed by detecting the tumor inhibition rate. Results: The recombinant fowlpox virus vFVHN was successfully constructed and its killing rate on SMMC7721 cells was 43.37%. The inhibitory rate of vFVHN on C57BL / 6 mice liver cancer model was 20.65%. Conclusion: The recombinant fowlpox virus vFVHN can effectively kill human hepatoma SMMC7721 cultured in vitro and inhibit the growth of solid tumors in C57BL / 6 mice.