携带CD自杀基因NDV重组病毒对小鼠肝癌细胞转移模型的治疗效果

采用反向遗传操作技术构建新城疫病毒基因组NP/P位点表达胞嘧啶脱氨酶(Cytosine deaminase,CD)基因弱毒力重组新城疫病毒rClone30-CD和中毒力重组新城疫病毒rAnhinga-CD。MTT检测结果表明,在协同5-氟胞嘧啶(5-Fluorocytosine,5-FC)条件下,两株重组新城疫病毒可在体外有效杀伤人肝癌细胞HepG2。通过Balb/c鼠尾静脉注射小鼠肝癌细胞H22构建小鼠肝癌细胞转移模型,经重组病毒协同5-FC联合治疗,荷瘤小鼠转移肿瘤体积显著缩小、肿瘤组织出现明显凋亡与坏死。体内试验结果表明,中毒力重组新城疫病毒rAnhinga-CD抑瘤效果优于弱毒力重组新城疫病毒r Clone30-CD。

Therapeutic effect of NDV recombinant virus carrying CD suicide gene on metastasis model of mouse hepatocellular carcinoma cells

The recombinant newcastle disease virus(rNDV) lentogenic strain rClone30-CD and velogenic strain rAnhinga-CD expressing cytosine deaminase(CD) gene at NP/P site in NDV genome were rescued by reverse genetic manipulation. MTT assay was used to detect the inhibitory effect of rClone30-CD and r Anhinga-CD combined with 5-Fluorocytosine(5-FC) on HepG2 cells in vitro. The results showed that the two recombinant viruses combined with 5-FC could kill HepG2 cells in a dosedependent manner. To prove the inhibitory effect of recombinant newcastle disease virus(rNDV)combined with CD/5-FC system on metastasis of hepatocellular carcinoma in mice, a model of H22 metastasis of hepatocellular carcinoma in Balb/c mice was established. After rNDVs treatment combinating of CD/5-FC system, the metastatic tumors in each group were weighed and the tumor tissue slices were stained with HE and TUNEL to evaluate the therapeutic effect of each group. The weight of metastasis tumors was significantly reduced and the obvious apoptosis and necrosis were found in tumor tissue. The inhibit effect of rAnhinga-CD was more efficient.