重组新城疫病毒rNDV-IL15在黑色素瘤治疗中的效果  

Antitumor efficacy of the recombinant Newcastle disease virus rNDV-IL15 on melanoma models

作　　者：牛泽杉[1] 白福良[1] 孙田[1] 田辉[1] 尹杰超[1] 曹宏伟[2] 于丹[1] 田贵游[1] 吴云舟[1] 李德山[1] 任桂萍[1] 

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]黑龙江八一农垦大学生命科学学院,黑龙江大庆163319

出　　处：《药学学报》2014年第3期310-315,共6页Acta Pharmaceutica Sinica

基　　金：国家自然基金东北农业大学生物学理科基地科研训练及科研能力提高项目(J1210069/J0106)

摘　　要：为了提高重组新城疫病毒抑制肿瘤的效果,本实验拯救获得了重组新城疫病毒rNDV-IL15。构建prNDV-IL15重组质粒,转染BHK21细胞后,拯救重组病毒rNDV-IL15,并测定病毒生长曲线;rNDV-IL15以0.1 MOI感染B16F10细胞,ELISA法检测细胞上清液中IL15的表达量;MTT法比较rNDV-IL15和rNDV在体外抑制B16F10细胞的效果,并比较了两者在黑色素瘤肿瘤模型中对荷瘤小鼠的治疗效果。结果表明,rNDV-IL15构建并成功拯救;病毒生长曲线表明,插入IL15后对病毒的生长无影响;IL15在细胞上清液中有较高的表达,达到(1 044.3±27.7)ng·mL-1;rNDV-IL15和rNDV对B16F10细胞的抑制率呈时间依赖性,但两者的抑制率无显著差异;与rNDV相比,rNDV-IL15在体内能较明显地抑制肿瘤生长。结果提示,rNDV-IL15是一种更有效的抗肿瘤制剂。In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of ILl5 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of ILl5 gene. Results showed that there was high level of ILl5 expression in the supernatant of rNDV-ILS-infected B16F10 cells （1 044.3 ± 27.7 ng·mL^-1）, rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.

关 键 词：重组新城疫病毒 IL15 rNDV-IL15 黑色素瘤 B16F10 抗肿瘤制剂 

分 类 号：R963[医药卫生—微生物与生化药学]