表达新城疫病毒HN基因重组鸡痘病毒的构建及其抑瘤作用

Construction of recombinant fowl poxvirus harboring HN gene of newcastle disease virus and its anti-tumor effect

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 establish a recombinant fowlpox virus vector harboring HN gene of Newcastle disease virus and to study its inhibitory effect on in vitro cultured human hepatic carcinoma cell line SMMC7721 and on H22 hepatoma in C57BL/6 mice. Methods： The fowl poxvirus 282 FA was transfected with HN gene of Newcastle disease virus, and the product （vFVHN） was screened by 5-Bromo-2-deoxyribouridine and identified by RT-PCR and Western blotting. The mortality of SMMC7721 cells was detected by Methyhhiazoletetra-zolium staining after vFVHN infection ; the tumor suppression rate of vFVHN＇on H22 hepatoma in C57BL/6 mice was detected to study its anti-tumor effect. Results： Fowlpox virus harboring HN gene of Newcastle disease was successfully constructed, and its killing effect on SMMC7721 was 43. 37% and its anti-tumor rate on SMMC7721 cells was 20.65%. Conclusion： It is demonstrated that vFVHN can effectively kill SMMC7721 cells cultured in vitro and has inhibitory effect on C57BL/6 mice bearing H22 hepatoma.