重组新城疫病毒对肺癌荷瘤鼠的治疗效果研究  

Study on therapeutic effect of recombinant Newcastle disease virus on lung cancer-bearing mice

作　　者：李德山[1] 张旭[1] 刘云野[1] 田贵游[1] 孙田[1] 安莹[1] 陈睿[1] LI Deshan;ZHANG Xu;LIU Yunye;TIAN Guiyou;SUN Tian;ANYing;CHEN Rui(School of Life Sciences, Northeast Agricultural University, Harbin 150030, China)

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2018年第11期1-8,17,共9页Journal of Northeast Agricultural University

基　　金：国家重点研发计划项目(2017YFD050103-03)

摘　　要：新城疫病毒用于肿瘤治疗具有良好靶向性和安全性,在众多溶瘤制剂中有独特优势。试验通过反向遗传操作技术成功拯救3株重组新城疫病毒r Clone30-IL2、r Clone30-P53和r Clone30-P53-IL2。经RT-PCR鉴定,表明插入新城疫病毒基因组中IL2、P53基因和P53-IL2双基因均插入预期位置,未发生突变。重组新城疫病毒在DF-1细胞中生长曲线表明,其与原代毒株生长特征相同。重组病毒在鸡胚中增殖稳定性试验表明,外源基因插入未影响重组病毒在鸡胚中增殖能力。体外肿瘤细胞生长抑制试验表明,与空载体r Clone30相比,r Clone30-IL2、r Clone30-P53和r Clone30-P53-IL2对LLC肺肿瘤细胞生长抑制效果更显著(P<0.01),呈剂量依赖性。体内抑瘤试验结果表明,r Clone30-IL2、r Clone30-P53和r Clone30-P53-IL2与尿囊液相比对肿瘤抑制效果差异极显著(P<0.01)。r Clone30-P53-IL2与r Clone30-IL2和rClone30-P53相比,对肿瘤抑制效果差异显著(P<0.05)。小鼠生存率结果表明,经rClone30-P53-IL-2治疗小鼠存活率最高,与其他组相比差异极显著(P <0.01)。综上所述,联合P53和IL2重组新城疫病毒对肺肿瘤治疗效果优于单一抗肿瘤相关因子。研究为筛选先导病毒治疗肺癌奠定基础。The advantage by using Newcastle disease virus for tumor therapy is tumor specific and safe.In this study,three recombinant Newcastle disease virus strains,rClone30-Il2,rClone30-P53and rClone30-P53-Il2,were successfully saved by reverse genetic manipulation The sequencing results showed that IL2,P53and p53-IL2genes were inserted in right location and not mutated.The growth curve experiment of recombinant Newcastle disease viruses in DF-1cells showed that the recombinant Newcastle disease viruses had the same growth characteristics as the wild type strain,suggesting that the insertion of exogenous genes did not affect the proliferation of recombinant viruses in chicken embryos.In vitro tumor suppressive experiment results showed that rClone30-P53and rClone30-P53-IL2had significant growth inhibition effects on LLC lung tumor cells(P<0.01)in a dose-dependent manner compared with rClone30and rClone30-IL2.In vivo tumor inhibition experiment,results showed that the inhibition effect of rClone30-P53-IL2was more potent than that of rClone30(P<0.01),rClone30-IL2and rClone30-P53(P<0.05).The results of the survival rate showed that the mice treated with rClone30-P53-IL-2had the longest survival rate(P<0.01).To summary,recombinant Newcastle disease virus simultaneously expressing P53and IL2had better tumor inhibition than those with single factors.This study laid a foundation for screening the lead virus for the treatment of lung cancer.

关 键 词：重组新城疫病毒 反向遗传操作技术 肺肿瘤 

分 类 号：R734.2[医药卫生—肿瘤]