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

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.