新城疫病毒复制对其溶瘤作用影响的初步分析

Role of Viral Replication in the Oncolysis of Newcastle Disease Virus

本研究旨在明确新城疫病毒(NDV)溶瘤作用是否依赖其复制水平,并改进NDV溶瘤机制研究模型。以NDV疫苗株LaSota感染人肿瘤细胞系HCT116、A549和人非肿瘤细胞系HEK293T为研究模型;RT-qPCR检测NDV在各细胞系的基因组复制水平;Western blot检测NDV在各细胞系的蛋白表达水平;利用流式细胞术检测NDV感染对各细胞系的细胞周期和细胞凋亡的影响。结果表明,NDV在三个细胞系内的基因组复制水平和蛋白表达水平没有显著差异,但对三个细胞系的溶瘤作用存在明显差异;进一步流式细胞术检测发现,NDV感染能诱发HEK293T和HCT116的细胞周期发生G1期停滞,但对A549的细胞周期没有明显影响;此外,NDV感染24 h后,A549细胞以早期凋亡为主,而HCT116细胞以坏死/晚期细胞凋亡为主。NDV的溶瘤作用并不依赖于其复制水平,而且NDV对不同类型肿瘤细胞的溶瘤作用存在不同机制。

The aim of this study was to determine the effect of the replication level of Newcastle disease virus(NDV)on its oncolytic effect and to improve the research model for studying oncolytic mechanism of NDV.The human tumor cell line HCT116,A549 and the human non-tumor cell line HEK293 T were infected with the NDV vaccine strain LaSota;the replication levels of viral genome in above mentioned cell lines were detected by NDV specific RT-qPCR;the expression levels of viral protein in above mentioned cell lines were assayed by Western blot;the effects of NDV infection on cell cycle and apoptosis of above mentioned cell lines were investigated using flow cytometry.The results revealed similar levels of viral replication but distinct oncolytic effects of NDV among these three cell lines;further flow cytometry showed that NDV infection induced G1 phase arrest in HEK293 T and HCT116 but not in A549;in addition,NDV mainly induced early apoptosis in A549 but necrosis/late apoptosis in HCT116.The oncolytic effect of NDV does not depend on its replication levels,and NDV kills human tumor cells by cell type-specific mechanisms.